Nucleic Acid Amplification Techniques
Sensitivity and Specificity
Polymerase Chain Reaction
Gene Amplification
Self-Sustained Sequence Replication
Ligase Chain Reaction
Gonorrhea
Neisseria gonorrhoeae
Keratin-19
DNA Primers
Base Sequence
Urine
Clinical significance of expression of human cytomegalovirus pp67 late transcript in heart, lung, and bone marrow transplant recipients as determined by nucleic acid sequence-based amplification. (1/1844)
Human cytomegalovirus (HCMV) infection was monitored retrospectively by qualitative determination of pp67 mRNA (a late viral transcript) by nucleic acid sequence-based amplification (NASBA) in a series of 50 transplant recipients, including 26 solid-organ (11 heart and 15 lung) transplant recipients (SOTRs) and 24 bone marrow transplant recipients (BMTRs). NASBA results were compared with those obtained by prospective quantitation of HCMV viremia and antigenemia and retrospective quantitation of DNA in leukocytes (leukoDNAemia). On the whole, 29 patients were NASBA positive, whereas 10 were NASBA negative, and the blood of 11 patients remained HCMV negative. NASBA detected HCMV infection before quantitation of viremia did but after quantitation of leukoDNAemia and antigenemia did. In NASBA-positive blood samples, median levels of viremia, antigenemia, and leukoDNAemia were significantly higher than the relevant levels detected in NASBA-negative HCMV-positive blood samples. By using the quantitation of leukoDNAemia as the "gold standard," the analytical sensitivity (47.3%), as well as the negative predictive value (68. 3%), of NASBA for the diagnosis of HCMV infection intermediate between that of antigenemia quantitation (analytical sensitivity, 72. 3%) and that of viremia quantitation (analytical sensitivity, 28.7%), while the specificity and the positive predictive value were high (90 to 100%). However, with respect to the clinically relevant antigenemia cutoff of >/=100 used in this study for the initiation of preemptive therapy in SOTRs with reactivated HCMV infection, the clinical sensitivity of NASBA reached 100%, with a specificity of 68. 9%. Upon the initiation of antigenemia quantitation-guided treatment, the actual median antigenemia level was 158 (range, 124 to 580) in SOTRs who had reactivated infection and who presented with NASBA positivity 3.5 +/- 2.6 days in advance and 13.5 (range, 1 to 270) in the group that included BMTRs and SOTRs who had primary infection (in whom treatment was initiated upon the first confirmation of detection of HCMV in blood) and who presented with NASBA positivity 2.0 +/- 5.1 days later. Following antiviral treatment, the durations of the presence of antigenemia and pp67 mRNA in blood were found to be similar. In conclusion, monitoring of the expression of HCMV pp67 mRNA appears to be a promising, well-standardized tool for determination of the need for the initiation and termination of preemptive therapy. Its overall clinical impact should be analyzed in future prospective studies. (+info)Comparison of levels of human immunodeficiency virus type 1 RNA in plasma as measured by the NucliSens nucleic acid sequence-based amplification and Quantiplex branched-DNA assays. (2/1844)
This study compared levels of human immunodeficiency virus type 1 RNA in plasma as measured by the Quantiplex branched-DNA and NucliSens nucleic acid sequence-based amplification assays. RNA was detectable in 118 of 184 samples (64.13%) by the Quantiplex assay and in 171 of 184 samples (92.94%) by the NucliSens assay. Regression analysis indicated that a linear relationship existed between the two sets of values (P < 0.0001), although the Quantiplex and NucliSens values were significantly different (P < 0.001), with the NucliSens values being approximately 0.323 log higher. Spearman correlation analysis indicated that the overall changes in patient viral load patterns were highly correlative between the two assays: r = 0.912, P < 0.0001. The lower limits of sensitivity were determined to be approximately 100 copies/ml and 1,200 to 1,400 copies/ml for the NucliSens and Quantiplex assays, respectively. (+info)Prospective comparison of the Gen-probe PACE 2 assay and the Abbott ligase chain reaction for the direct detection of Chlamydia trachomatis in a low prevalence population. (3/1844)
In a prospective study, the Gen-Probe PACE 2 (GP) assay was compared with Abbott Laboratories' ligase chain reaction (LCR) assay for the detection of Chlamydia trachomatis. A total of 493 female patients consented to collection of two cervical samples; a first-void urine (FVU) sample was collected also from 446 of the participants. Cervical samples were tested by both GP and LCR; 16 samples (3.1%) tested positive by both methods and no discrepant results were observed. All but one of the FVU samples collected from patients with a positive cervical sample was positive for C. trachomatis by LCR. The stability of FVU samples over time in the LCR test was also evaluated and proved to be significantly longer than the 4 days stated by the manufacturer. While LCR proved to be highly sensitive in detecting chlamydial infection in FVU samples, no difference was noted between LCR and GP in the detection of cervical C. trachomatis infection in this study population. (+info)Design and evaluation of a human immunodeficiency virus type 1 RNA assay using nucleic acid sequence-based amplification technology able to quantify both group M and O viruses by using the long terminal repeat as target. (4/1844)
Currently available human immunodeficiency virus type 1 (HIV-1) RNA quantification assays can detect most viruses of the group M subtypes, but a substantial number are missed or not quantified reliably. Viruses of HIV-1 group O cannot be detected by any commercially available assay. We developed and evaluated a quantitative assay based on nucleic acid sequence-based amplification (NASBA) technology, with primers and probes located in the conserved long terminal repeat (LTR) region of the HIV-1 genome. In 68 of 72 serum samples from individuals infected with HIV-1 subtypes A to H of group M, viruses could be detected and quantified. In serum samples from two patients infected with HIV-1 group O viruses, these viruses as well could be detected and quantified. In contrast, the currently used gag-based assay underestimated the presence of subtype A viruses and could not detect subtype G and group O viruses. The discrepancy between the results of the two assays may be explained by the number of mismatches found within and among the probe and primer regions of the subtype isolates. These data indicate that LTR-based assays, including the NASBA format chosen here, are better suited to monitoring HIV-1 therapy than are gag-based assays in an era in which multiple HIV-1 subtypes and groups are spreading worldwide. (+info)High-resolution genotyping of Streptococcus pyogenes serotype M1 isolates by fluorescent amplified-fragment length polymorphism analysis. (5/1844)
We have used fluorescent amplified-fragment length polymorphism (FAFLP) analysis to subtype clinical isolates of Streptococcus pyogenes serotype M1. Established typing methods define most M1 isolates as members of a clone that has a worldwide distribution and that is strongly associated with invasive diseases. FAFLP analysis simultaneously sampled 90 to 120 loci throughout the M1 genome. Its discriminatory power, precision, and reproducibility were compared with those of other molecular typing methods. Irrespective of disease symptomatology or geographic origin, the majority of the clinical M1 isolates shared a single ribotype, pulsed-field gel electrophoresis macrorestriction profile, and emm1 gene sequence. Nonetheless, among these isolates, FAFLP analysis could differentiate 17 distinct profiles, including seven multi-isolate groups. The FAFLP profiles of M1 isolates reproducibly exhibited between 1 and more than 20 amplified fragment differences. The high discriminatory power of genotyping by FAFLP analysis revealed genetic microheterogeneity and differentiated otherwise "identical" M1 isolates as members of a clone complex. (+info)Amplified-fragment length polymorphism analysis versus macro-restriction fragment analysis for molecular typing of Streptococcus pneumoniae isolates. (6/1844)
Forty-eight pneumococci were genotyped by on-line laser fluorescence amplified-fragment length polymorphism (AFLP) and pulsed-field gel electrophoresis (PFGE) analysis of chromosomal restriction fragments. Overall, the data generated by the two methods corresponded well. However, with AFLP, clusters were delineated at a higher similarity level, and isolate differentiation was more pronounced. AFLP and PFGE were equally efficient for assessing intraserotype diversity. We conclude that AFLP is a useful alternative to PFGE. (+info)Loss of heterozygosity analysis using whole genome amplification, cell sorting, and fluorescence-based PCR. (7/1844)
Loss of heterozygosity (LOH) is a common genetic lesion found in many human neoplasms. Extending investigation of LOH to large-scale clinical and public health science studies has proven difficult because of the small size and cellular and genetic heterogeneity of human neoplasms, in addition to the challenges associated with increasing throughput. Our approach to LOH analysis was developed using clinical biopsy samples from patients with Barrett's esophagus (BE) and uses flow cytometric cell sorting to increase sample purity, whole genome amplification to increase sample amount, and automated fluorescent genotyping to increase sample throughput. This approach allows LOH assessment at 20 loci in DNA extracted from 1000 flow-purified cells while maintaining accurate and reproducible allele ratios compared with the standard method of using genomic DNA. This method of analysis should allow accurate, reproducible determination of allele ratios in a variety of human tumors and premalignant conditions. (+info)Comparison of the PACE 2 assay, two amplification assays, and Clearview EIA for detection of Chlamydia trachomatis in female endocervical and urine specimens. (8/1844)
Screening for sexually transmitted diseases (STDs) in a greater proportion of sexually active patients has become an accepted protocol by most health care providers. The purpose of this study was to compare the current test methods for detection of Chlamydia trachomatis used at the University of South Alabama, the PACE 2 assay (Gen-Probe) and the Clearview EIA (Wampole Laboratories), with two amplification technologies, the AMP CT (Gen-Probe) and LCx (Abbott) assays. In addition, a number of demographic parameters were ascertained by asking questions at the time of examination as well as for health care provider concerns and preferences. One urine and four endocervical swab specimens were collected in random order from 787 female patients attending one of four obstetrics-gynecology clinics. Eighty-seven percent of patients had no STD-related symptoms. Patients were considered positive for C. trachomatis if three or more assays (swab and/or urine) were positive. Abbott and Gen-Probe confirmed discrepant results by alternate amplified assays. A total of 66 true-positive specimens were detected by use of the combination of endocervical swabs and urine specimens. After discrepant analysis, sensitivities for endocervical swab specimens for the EIA and the PACE 2, LCx, and AMP CT assays were 50, 81, 97, and 100%, respectively. Sensitivities for the LCx and AMP CT assays with urine specimens were 98 and 81%, respectively. The prevalence of C. trachomatis was 8.4%, as determined by amplification technology. Overall, the amplification technologies were the most sensitive methods with either swab (AMP CT assay) or urine (LCx assay) specimens. The PACE 2 assay offered the advantage of a simpler and less expensive assay with acceptable sensitivity. The clearview CT EIA, while yielding a rapid in-office result, had unacceptably low sensitivity. The wide variation in performance with amplification assays with urine specimens as reported in both this study and the literature obviates the need to clarify optimal parameters for this specimen type. (+info)Nucleic acid amplification techniques (NAATs) are medical laboratory methods used to increase the number of copies of a specific DNA or RNA sequence. These techniques are widely used in molecular biology and diagnostics, including the detection and diagnosis of infectious diseases, genetic disorders, and cancer.
The most commonly used NAAT is the polymerase chain reaction (PCR), which involves repeated cycles of heating and cooling to separate and replicate DNA strands. Other NAATs include loop-mediated isothermal amplification (LAMP), nucleic acid sequence-based amplification (NASBA), and transcription-mediated amplification (TMA).
NAATs offer several advantages over traditional culture methods for detecting pathogens, including faster turnaround times, increased sensitivity and specificity, and the ability to detect viable but non-culturable organisms. However, they also require specialized equipment and trained personnel, and there is a risk of contamination and false positive results if proper precautions are not taken.
Sensitivity and specificity are statistical measures used to describe the performance of a diagnostic test or screening tool in identifying true positive and true negative results.
* Sensitivity refers to the proportion of people who have a particular condition (true positives) who are correctly identified by the test. It is also known as the "true positive rate" or "recall." A highly sensitive test will identify most or all of the people with the condition, but may also produce more false positives.
* Specificity refers to the proportion of people who do not have a particular condition (true negatives) who are correctly identified by the test. It is also known as the "true negative rate." A highly specific test will identify most or all of the people without the condition, but may also produce more false negatives.
In medical testing, both sensitivity and specificity are important considerations when evaluating a diagnostic test. High sensitivity is desirable for screening tests that aim to identify as many cases of a condition as possible, while high specificity is desirable for confirmatory tests that aim to rule out the condition in people who do not have it.
It's worth noting that sensitivity and specificity are often influenced by factors such as the prevalence of the condition in the population being tested, the threshold used to define a positive result, and the reliability and validity of the test itself. Therefore, it's important to consider these factors when interpreting the results of a diagnostic test.
Polymerase Chain Reaction (PCR) is a laboratory technique used to amplify specific regions of DNA. It enables the production of thousands to millions of copies of a particular DNA sequence in a rapid and efficient manner, making it an essential tool in various fields such as molecular biology, medical diagnostics, forensic science, and research.
The PCR process involves repeated cycles of heating and cooling to separate the DNA strands, allow primers (short sequences of single-stranded DNA) to attach to the target regions, and extend these primers using an enzyme called Taq polymerase, resulting in the exponential amplification of the desired DNA segment.
In a medical context, PCR is often used for detecting and quantifying specific pathogens (viruses, bacteria, fungi, or parasites) in clinical samples, identifying genetic mutations or polymorphisms associated with diseases, monitoring disease progression, and evaluating treatment effectiveness.
Molecular diagnostic techniques are a group of laboratory methods used to analyze biological markers in DNA, RNA, and proteins to identify specific health conditions or diseases at the molecular level. These techniques include various methods such as polymerase chain reaction (PCR), DNA sequencing, gene expression analysis, fluorescence in situ hybridization (FISH), and mass spectrometry.
Molecular diagnostic techniques are used to detect genetic mutations, chromosomal abnormalities, viral and bacterial infections, and other molecular changes associated with various diseases, including cancer, genetic disorders, infectious diseases, and neurological disorders. These techniques provide valuable information for disease diagnosis, prognosis, treatment planning, and monitoring of treatment response.
Compared to traditional diagnostic methods, molecular diagnostic techniques offer several advantages, such as higher sensitivity, specificity, and speed. They can detect small amounts of genetic material or proteins, even in early stages of the disease, and provide accurate results with a lower risk of false positives or negatives. Additionally, molecular diagnostic techniques can be automated, standardized, and performed in high-throughput formats, making them suitable for large-scale screening and research applications.
Gene amplification is a process in molecular biology where a specific gene or set of genes are copied multiple times, leading to an increased number of copies of that gene within the genome. This can occur naturally in cells as a response to various stimuli, such as stress or exposure to certain chemicals, but it can also be induced artificially through laboratory techniques for research purposes.
In cancer biology, gene amplification is often associated with tumor development and progression, where the amplified genes can contribute to increased cell growth, survival, and drug resistance. For example, the overamplification of the HER2/neu gene in breast cancer has been linked to more aggressive tumors and poorer patient outcomes.
In diagnostic and research settings, gene amplification techniques like polymerase chain reaction (PCR) are commonly used to detect and analyze specific genes or genetic sequences of interest. These methods allow researchers to quickly and efficiently generate many copies of a particular DNA sequence, facilitating downstream analysis and detection of low-abundance targets.
"Self-Sustained Sequence Replication" is not a recognized medical term. It appears to be related to the field of molecular biology, specifically in the study of DNA replication and gene expression. However, I am an assistant trained to assist with general knowledge questions and not a medical professional. Therefore, I would recommend consulting a reliable medical source or speaking with a healthcare provider for accurate information regarding this term.
Ligase Chain Reaction (LCR) is a highly specific and sensitive method used in molecular biology for the detection of point mutations or small deletions or insertions in DNA. It is an enzymatic reaction-based technique that relies on the repeated ligation of adjacent oligonucleotide probes to form a continuous strand, followed by thermal denaturation and reannealing of the strands.
The LCR process involves the use of a thermostable ligase enzyme, which catalyzes the formation of a phosphodiester bond between two adjacent oligonucleotide probes that are hybridized to complementary sequences in the target DNA. The oligonucleotides are designed to have a gap at the site of the mutation or deletion/insertion, such that ligation can only occur if the probe sequences match perfectly with the target DNA.
After each round of ligation and denaturation, the reaction mixture is subjected to PCR amplification using primers flanking the region of interest. The amplified products are then analyzed for the presence or absence of ligated probes, indicating the presence or absence of the mutation or deletion/insertion in the target DNA.
LCR has been widely used in diagnostic and research applications, including the detection of genetic diseases, infectious agents, and cancer-associated mutations. However, it has largely been replaced by other more sensitive and high-throughput methods such as real-time PCR and next-generation sequencing.
'Chlamydia trachomatis' is a species of bacterium that is the causative agent of several infectious diseases in humans. It is an obligate intracellular pathogen, meaning it can only survive and reproduce inside host cells. The bacteria are transmitted through sexual contact, and can cause a range of genital tract infections, including urethritis, cervicitis, pelvic inflammatory disease, and epididymitis. In women, chlamydial infection can also lead to serious complications such as ectopic pregnancy and infertility.
In addition to genital infections, 'Chlamydia trachomatis' is also responsible for two other diseases: trachoma and lymphogranuloma venereum (LGV). Trachoma is a leading cause of preventable blindness worldwide, affecting mostly children in developing countries. It is spread through contact with contaminated hands, clothing, or eye secretions. LGV is a sexually transmitted infection that can cause inflammation of the lymph nodes, rectum, and genitals.
'Chlamydia trachomatis' infections are often asymptomatic, making them difficult to diagnose and treat. However, they can be detected through laboratory tests such as nucleic acid amplification tests (NAATs) or culture. Treatment typically involves antibiotics such as azithromycin or doxycycline. Prevention measures include safe sex practices, regular screening for STIs, and good hygiene.
Chlamydia infections are caused by the bacterium Chlamydia trachomatis and can affect multiple body sites, including the genitals, eyes, and respiratory system. The most common type of chlamydia infection is a sexually transmitted infection (STI) that affects the genitals.
In women, chlamydia infections can cause symptoms such as abnormal vaginal discharge, burning during urination, and pain in the lower abdomen. In men, symptoms may include discharge from the penis, painful urination, and testicular pain or swelling. However, many people with chlamydia infections do not experience any symptoms at all.
If left untreated, chlamydia infections can lead to serious complications, such as pelvic inflammatory disease (PID) in women, which can cause infertility and ectopic pregnancy. In men, chlamydia infections can cause epididymitis, an inflammation of the tube that carries sperm from the testicles, which can also lead to infertility.
Chlamydia infections are diagnosed through a variety of tests, including urine tests and swabs taken from the affected area. Once diagnosed, chlamydia infections can be treated with antibiotics such as azithromycin or doxycycline. It is important to note that treatment only clears the infection and does not repair any damage caused by the infection.
Prevention measures include practicing safe sex, getting regular STI screenings, and avoiding sharing towels or other personal items that may come into contact with infected bodily fluids.
Gonorrhea is a sexually transmitted infection (STI) caused by the bacterium Neisseria gonorrhoeae, also known as "gono" bacteria. It can infect various parts of the body including the genitals, rectum, and throat. The bacteria are typically transmitted through sexual contact with an infected person.
Symptoms may vary but often include abnormal discharge from the genitals or rectum, painful or burning sensations during urination, and in women, vaginal bleeding between periods. However, many people with gonorrhea do not develop symptoms, making it essential to get tested regularly if you are sexually active with multiple partners or have unprotected sex.
If left untreated, gonorrhea can lead to severe complications such as pelvic inflammatory disease (PID) in women and epididymitis in men, which may result in infertility. In rare cases, it can spread to the bloodstream and cause life-threatening conditions like sepsis.
Gonorrhea is curable with appropriate antibiotic treatment; however, drug-resistant strains of the bacteria have emerged, making accurate diagnosis and effective treatment increasingly challenging. Prevention methods include using condoms during sexual activity and practicing safe sex habits.
Neisseria gonorrhoeae is a species of gram-negative, aerobic diplococcus that is the etiologic agent of gonorrhea, a sexually transmitted infection. It is commonly found in the mucous membranes of the reproductive tract, including the cervix, urethra, and rectum, as well as the throat and eyes. The bacterium can cause a range of symptoms, including discharge, burning during urination, and, in women, abnormal menstrual bleeding. If left untreated, it can lead to more serious complications, such as pelvic inflammatory disease and infertility. It is important to note that N. gonorrhoeae has developed resistance to many antibiotics over time, making treatment more challenging. A culture or nucleic acid amplification test (NAAT) is used for the diagnosis of this infection.
Bacterial DNA refers to the genetic material found in bacteria. It is composed of a double-stranded helix containing four nucleotide bases - adenine (A), thymine (T), guanine (G), and cytosine (C) - that are linked together by phosphodiester bonds. The sequence of these bases in the DNA molecule carries the genetic information necessary for the growth, development, and reproduction of bacteria.
Bacterial DNA is circular in most bacterial species, although some have linear chromosomes. In addition to the main chromosome, many bacteria also contain small circular pieces of DNA called plasmids that can carry additional genes and provide resistance to antibiotics or other environmental stressors.
Unlike eukaryotic cells, which have their DNA enclosed within a nucleus, bacterial DNA is present in the cytoplasm of the cell, where it is in direct contact with the cell's metabolic machinery. This allows for rapid gene expression and regulation in response to changing environmental conditions.
Keratin-19 is a type I acidic keratin that is primarily expressed in simple epithelia, such as the gastrointestinal tract, respiratory tract, and epidermal appendages (e.g., hair follicles, sweat glands). It plays an essential role in maintaining the structure and integrity of these tissues by forming intermediate filaments that provide mechanical support to cells.
Keratin-19 is often used as a marker for simple epithelial differentiation and has been implicated in various pathological conditions, including cancer progression and metastasis. Mutations in the KRT19 gene, which encodes keratin-19, have been associated with certain genetic disorders, such as epidermolysis bullosa simplex, a blistering skin disorder.
In summary, Keratin-19 is an important structural protein expressed in simple epithelia that plays a crucial role in maintaining tissue integrity and has implications in various pathological conditions.
DNA primers are short single-stranded DNA molecules that serve as a starting point for DNA synthesis. They are typically used in laboratory techniques such as the polymerase chain reaction (PCR) and DNA sequencing. The primer binds to a complementary sequence on the DNA template through base pairing, providing a free 3'-hydroxyl group for the DNA polymerase enzyme to add nucleotides and synthesize a new strand of DNA. This allows for specific and targeted amplification or analysis of a particular region of interest within a larger DNA molecule.
A base sequence in the context of molecular biology refers to the specific order of nucleotides in a DNA or RNA molecule. In DNA, these nucleotides are adenine (A), guanine (G), cytosine (C), and thymine (T). In RNA, uracil (U) takes the place of thymine. The base sequence contains genetic information that is transcribed into RNA and ultimately translated into proteins. It is the exact order of these bases that determines the genetic code and thus the function of the DNA or RNA molecule.
Bacteriological techniques refer to the various methods and procedures used in the laboratory for the cultivation, identification, and study of bacteria. These techniques are essential in fields such as medicine, biotechnology, and research. Here are some common bacteriological techniques:
1. **Sterilization**: This is a process that eliminates or kills all forms of life, including bacteria, viruses, fungi, and spores. Common sterilization methods include autoclaving (using steam under pressure), dry heat (in an oven), chemical sterilants, and radiation.
2. **Aseptic Technique**: This refers to practices used to prevent contamination of sterile materials or environments with microorganisms. It includes the use of sterile equipment, gloves, and lab coats, as well as techniques such as flaming, alcohol swabbing, and using aseptic transfer devices.
3. **Media Preparation**: This involves the preparation of nutrient-rich substances that support bacterial growth. There are various types of media, including solid (agar), liquid (broth), and semi-solid (e.g., stab agar). The choice of medium depends on the type of bacteria being cultured and the purpose of the investigation.
4. **Inoculation**: This is the process of introducing a bacterial culture into a medium. It can be done using a loop, swab, or needle. The inoculum should be taken from a pure culture to avoid contamination.
5. **Incubation**: After inoculation, the bacteria are allowed to grow under controlled conditions of temperature, humidity, and atmospheric composition. This process is called incubation.
6. **Staining and Microscopy**: Bacteria are too small to be seen with the naked eye. Therefore, they need to be stained and observed under a microscope. Gram staining is a common method used to differentiate between two major groups of bacteria based on their cell wall composition.
7. **Biochemical Tests**: These are tests used to identify specific bacterial species based on their biochemical characteristics, such as their ability to ferment certain sugars, produce particular enzymes, or resist certain antibiotics.
8. **Molecular Techniques**: Advanced techniques like PCR and DNA sequencing can provide more precise identification of bacteria. They can also be used for genetic analysis and epidemiological studies.
Remember, handling microorganisms requires careful attention to biosafety procedures to prevent accidental infection or environmental contamination.
Urine is a physiological excretory product that is primarily composed of water, urea, and various ions (such as sodium, potassium, chloride, and others) that are the byproducts of protein metabolism. It also contains small amounts of other substances like uric acid, creatinine, ammonia, and various organic compounds. Urine is produced by the kidneys through a process called urination or micturition, where it is filtered from the blood and then stored in the bladder until it is excreted from the body through the urethra. The color, volume, and composition of urine can provide important diagnostic information about various medical conditions.
Specimen handling is a set of procedures and practices followed in the collection, storage, transportation, and processing of medical samples or specimens (e.g., blood, tissue, urine, etc.) for laboratory analysis. Proper specimen handling ensures accurate test results, patient safety, and data integrity. It includes:
1. Correct labeling of the specimen container with required patient information.
2. Using appropriate containers and materials to collect, store, and transport the specimen.
3. Following proper collection techniques to avoid contamination or damage to the specimen.
4. Adhering to specific storage conditions (temperature, time, etc.) before testing.
5. Ensuring secure and timely transportation of the specimen to the laboratory.
6. Properly documenting all steps in the handling process for traceability and quality assurance.
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Polymerase Chain Reaction ProtocolPolymerase13
- The World Health Organization (WHO) reports that nucleic acid amplification tests, such as quantitative nucleic acid sequence-based amplification, polymerase chain reaction (PCR) loop-mediated isothermal amplification (LAMP), including quantitative or real-time PCR, are used to detect malaria infections as of February 2019. (medgadget.com)
- It is a quick polymerase chain reaction-related nucleic acid amplification technique. (medgadget.com)
- The polymerase chain reaction (PCR) is a technique that allows amplification of nucleic acids in-vitro. (biosyn.com)
- Furthermore, a variety of technological improvements, these as the improvement of Strand Displacement Amplification (SDA), Single Primer Isothermal Amplification (SPIA) and Recombinase Polymerase Amplification (RPA), is performing as an additional development-inducing factor. (floschi.info)
- Lyo-ready Bst DNA polymerase demonstrates fast viral RNA amplification with detection in as few as ten minutes in an RT-LAMP reaction, using SSIV reverse transcriptase for the initial amplification step. (thermofisher.com)
- The high sensitivity of Lyo-ready Bst DNA Polymerase enables amplification from a limited amount of starting material or amplification from samples containing low concentration of the target DNA/RNA. (thermofisher.com)
- As amounts of DNA and RNA in samples can be minute, the researchers applied a technique called recombinase polymerase amplification, which can amplify nucleic acids without special equipment. (nih.gov)
- Nucleic acid amplification, commonly known as polymerase chain reaction (PCR), is one of the most revolutionary developments in modern molecular biology. (strategic-directions.com)
- Further attempts to increase sensitivity and specificity of sentinel node analysis include molecular biology-based techniques such as the real-time polymerase chain reaction (RT-PCR) and, more recently, one step nucleic acid amplification (OSNA). (minervamedica.it)
- Common detection methods include enzyme-linked immunosorbent assay (ELISA), immunofluorescence, and nucleic acid amplification techniques (such as polymerase chain reaction, PCR). (wkinformation.com)
- Polymerase Chain Reaction (PCR) is a widely used technique, extensively applied in clinical diagnosis and molecular biology research. (mantacc.com)
- PCR is the amplification of specific nucleic acid (NA) sequences by DNA polymerase in vitro. (mantacc.com)
- PCR amplifies target nucleic acid sequences using DNA polymerase, primers, and nucleotides. (mantacc.com)
Loop-mediated isot5
- The GEAR technique is an improvement over loop-mediated isothermal amplification (LAMP) in three ways. (nih.gov)
- The global market for isothermal nucleic acid amplification technology (INAAT) has by product type instruments and reagents & kits, by technology loop-mediated isothermal amplification, helicase dependent amplification, single primer isothermal amplification, nicking enzyme amplification reaction, transcription-medicated amplification, and nucleic acid sequence-based amplification, and by application blood screening, and infectious disease diagnostics, is segmented from 2019 - 2027. (medgadget.com)
- In line with this, the widespread adoption of Loop-mediated Isothermal Amplification (LAMP) tests that are used for the amplification of both equally DNA and RNA, together with the identification of genetically modified organisms (GMOs) is driving the merchandise need further more. (floschi.info)
- Loop-mediated isothermal amplification, also known as LAMP, is a simple and cost-effective nucleic acid amplification technique that does not require expensive instruments and helps ensure rapid results and easy outcome interpretation. (thermofisher.com)
- The LAMP (loop-mediated isothermal amplification) test is a form of testing in which small fragments of genetic material are copied or replicated, similar to the PCR test. (rivm.nl)
Target nucleic acid7
- NEAA reactions offer high levels of amplification efficiency and can exponentially amplify a target nucleic acid over a very short period of time. (nih.gov)
- This technology employs photoinduced electron transfer (PET) nucleic acid molecules that can be used detect and amplify target nucleic acid molecules. (nih.gov)
- The universal application of PCR lies in its ability to amplify a small quantity of target nucleic acid sequences into a large amount of PCR products, which can then be detected by downstream methods, such as observing nucleic acid (NA) products through agarose gel electrophoresis. (mantacc.com)
- This is due to the exponential amplification of nucleic acid (NA) sequences, resulting in the original template (target nucleic acid sequence) being amplified to produce millions of copies. (mantacc.com)
- The template for a PCR reaction can be any target nucleic acid sequence. (mantacc.com)
- Primers are designed to complementarily bind to the antisense strand of a specific target nucleic acid template, with the length of the primers usually between 15-40 bases. (mantacc.com)
- This leads to exponential amplification of the original template, typically resulting in millions or billions of copies of the original target nucleic acid sequence. (mantacc.com)
Amplify1
- For instance, Genomtec received a Horizon 2020 SME phase I grant in December 2019 for Genomtec Tumor, a device that operates on the theory of streamlined nucleic acid amplification technology (SNAAT), which uses photon-based technology to heat the reaction mixture and amplify particular nucleic acid fragments that are specific to cancer. (medgadget.com)
Isothermal amplification2
- Nucleic acid isothermal amplification detection technologies have undergone rapid development in the field of molecular diagnosis due to their freedom from the limitations of complex temperature control equipment. (nih.gov)
- In recent years, with the use of newly discovered nicking endonucleases, a novel isothermal amplification technique, -nicking enzyme-assisted amplification (NEAA), has been developed. (nih.gov)
NAATs3
- and Nucleic Acid Amplification Techniques (NAATs). (wikipedia.org)
- Antigen tests, cell culture tests, routine screening, Nucleic Acid Amplification Tests (NAATs), and symptom based physical or biological examinations are generally conducted to determine the presence of Chlamydia infection. (streetinsider.com)
- Nucleic acid amplification techniques (NAATs) tend to be costly and in some cases lack sensitivity. (crimsonpublishers.com)
Assays3
- The performance of hepatitis E virus (HEV) RNA nucleic acid amplification (NAT)-based assays has been investigated using a panel of HEV-containing plasma samples. (nih.gov)
- As a result, the worldwide market for INAAT is anticipated to benefit from the use of nucleic acid assays in the diagnosis of malaria. (medgadget.com)
- Because of the improved culture methods and sensitive nucleic acid amplification assays developed in recent years, Kingella kingae , a gram-negative coccobacillus of the Neisseriaceae family, is increasingly recognized as an invasive pathogen of early childhood. (cdc.gov)
Transcription Mediated Amplification1
- For a TMA (Transcription Mediated Amplification) test, a sample is taken using the same method as the PCR test, and then processed in a laboratory. (rivm.nl)
Antigen5
- HIV infection can be diagnosed based on detection of antibodies that are directed against the proteins encoded by the 3 major genes, the detection of the p24 antigen, the viral nucleic acid, and, finally, by means of culturing the virus. (medscape.com)
- Due to the testing technique, the tests in the NAAT category are generally more sensitive than the rapid antigen tests (which includes self-tests). (rivm.nl)
- In biosensor-based diagnostic technique, an analytical device consisting of biological elements and physicochemical component gives a signal upon detection of viral antigen. (biomedcentral.com)
- In immunological-based diagnostic techniques, enzyme-linked antibodies are utilized to find the particular antiviral antibody or viral antigen in human specimens, and nucleic acid-based diagnostic techniques are based on the principle of amplification of the viral genome. (biomedcentral.com)
- The electrochemical biosensor is a device that contains bioreceptor elements which may be in a form of biocatalysts (enzymes, tissues) or affinity sensors (antibody, nucleic acid) that gets reacted with our target analyte which may be antibody-antigen, proteins, etc. (biomedcentral.com)
Sequences6
- The outer primer pair targets three specific nucleic acid sequences at a constant 60°C, while the inner pair of primers accelerates and improves the sensitivity of the assay. (nih.gov)
- The ability to quickly and cheaply detect minute amounts of specific nucleic acid (DNA and RNA) sequences could bring significant public health benefits. (nih.gov)
- At present, fluorescent primers are useful for detecting and identifying microbes and specific nucleic acid sequences, amplifying nucleic acids for pyro-sequencing, determining the levels of gene expression, and many other uses. (nih.gov)
- Laboratory techniques have been introduced for inducing disproportional replication by unequal crossing over, uptake of DNA from lysed cells, or generation of extrachromosomal sequences from rolling circle replication. (bvsalud.org)
- Primers are short sequences of nucleic acids synthesized in vitro. (mantacc.com)
- For example, amplification of very short target sequences requires much shorter durations at each temperature compared to very long target sequences. (mantacc.com)
Detection and Amplification1
- PCR enables the detection and amplification of a specific sequence, requiring only a single template to produce millions of copies. (strategic-directions.com)
NAAT3
- The category of tests known as NAAT (Nucleic Acid Amplification Test) includes the PCR test, the TMA test and the LAMP test. (rivm.nl)
- As the prevalence of influenza is increasing, the number of diagnostic techniques such as immunochromatography-based rapid diagnostic test (RDT) and nucleic acid amplification test (NAAT) are used for the detection of influenza viruses in humans. (medgadget.com)
- According to researchers, this new technique will cost $2.00 per test, while the Nucleic Acid Amplification Test (NAAT)-which is the current chlamydia-testing method being used-costs around $10.00 per test. (doctorshealthpress.com)
Viral2
- Amplification speed was determined in RT-LAMP reaction using SARS-CoV-2 synthetic RNA and Measles viral RNA, their respective primer sets, and SYTO 9 green fluorescent nucleic acid stain for real-time detection. (thermofisher.com)
- Blood testing devices: These devices are used to detect antibodies or viral nucleic acids in the blood to determine if you have been infected with a sexually transmitted disease, such as HIV or Syphilis. (wkinformation.com)
World Health Organization International S1
- The assay is calibrated to the First World Health Organization International Standard for EBV for nucleic acid amplification techniques. (testcatalog.org)
Molecules1
- Capillary electrophoresis is a family of analytical techniques that separate charged molecules based on their electrophoretic mobility through applied voltage. (strategic-directions.com)
Hybridization4
- Genotypic methods allow the direct detection of specific resistance genes, as well as any novel mutations that may have occurred ( Table 1 ), and tend to fall into one of three categories: amplification-based, sequence-based or hybridization-based methods. (integra-biosciences.com)
- Comprehensive chromosomal analysis of human preimplantation embryos using whole genome amplification and single cell comparative genomic hybridization. (ox.ac.uk)
- We have developed a novel technique based on whole genome amplification and comparative genomic hybridization (CGH), which for the first time allows the copy number of every chromosome to be assessed in almost every cell of a cleavage-stage embryo. (ox.ac.uk)
- Here, we report an innovative hybridization chain reaction (HCR) technique that involves computer-aided design (CAD) and reversible assembly. (cngb.org)
Specimens5
- On September 30, 1999, the Food and Drug Administration approved a reformulated Amplified Mycobacterium Tuberculosis Direct Test* (MTD) (Gen-Probe ® , San Diego, California) for detection of Mycobacterium tuberculosis in acid-fast bacilli (AFB) smear-positive and smear-negative respiratory specimens from patients suspected of having tuberculosis (TB). (cdc.gov)
- MTD and one other nucleic acid amplification (NAA) test, the Amplicor ® Mycobacterium Tuberculosis Test (Amplicor) (Roche ® Diagnostic Systems, Inc., Branchburg, New Jersey), previously had been approved for the direct detection of M. tuberculosis in respiratory specimens that have positive AFB smears. (cdc.gov)
- Selective Whole-Genome Amplification as a Tool to Enrich Specimens with Low Treponema pallidum Genomic DNA Copies for Whole-Genome Sequencing. (cdc.gov)
- Further fixation of specimens in GA containing ruthe nium red or tannic acid depicts that the interstitial interface inside of the renal stem progenitor cell niche consists of an unexpectedly substantial quantity of amorphous extracellular matrix. (agckinase.com)
- Sputum specimens subjected to fluorochrome staining is preferred because it can be done more rapidly, and it is a more sensitive technique to identify mycobacteria than carbolfuchsin staining. (japt.in)
Reaction9
- CDC researchers developed a simple target-specific isothermal nucleic acid amplification technique, termed Genome Exponential Amplification Reaction (GEAR). (nih.gov)
- To test for inhibitors of MTD, spike an aliquot of the lysated sputum sample with lysed M. tuberculosis (approximately 10 organisms per reaction, or an equivalent amount of M. tuberculosis rRNA) and repeat the test starting with amplification. (cdc.gov)
- The reaction is stopped by adding a sulphuric acid solution and the OD of each well is read. (cdc.gov)
- The reaction is stopped by adding a sulphuric acid solution and the OD of each well is read.The presence or absence of IgM antibodies to hepatitis E virus is determined by the ratio of the OD of each sample to the calculated cut-off value. (cdc.gov)
- The ligation-during-amplification (LDA) reaction can be used for the synthesis of circular oligonucleotides. (biosyn.com)
- Since the introduction of the ligation-during-amplification (LDA) reaction, in-vitro synthesis of closed circular DNA is possible without the need for subcloning. (biosyn.com)
- The final phase is the plateau phase, where the reaction stops and no more amplification products are generated. (mantacc.com)
- Phases of a PCR Reaction: During the PCR process, product doubling occurs in the exponential phase, product amplification slightly less than doubling occurs in the linear phase, and very low, almost stagnant product amplification occurs in the plateau phase. (mantacc.com)
- This signal indicates the amount of double-stranded nucleic acid contained in the reaction tube or well. (mantacc.com)
Sensitivity3
- High sensitivity and reliable amplification from low amounts of DNA. (thermofisher.com)
- The latter technique, that has sensitivity close to 100% and extremely high specificity along with good reproducibility, allows analysis of the SLN in full with an intraoperative procedure in approximately 30 minutes. (minervamedica.it)
- In situ spatial proteomics analysis of a single cell has not been achieved yet, mainly because of insufficient throughput and sensitivity of current techniques. (cngb.org)
Exponential amplification1
- The use of thermal cycling allows the exponential amplification of closed circular DNA. (biosyn.com)
Complementary2
- PET tag activity can be quenched by at least two consecutive guanosines (G-G) within the tag sequence and activity is un-quenched when the PET tag hybridizes with its complementary nucleic acid molecule. (nih.gov)
- The complementary room among the ruthenium red and tannic acid constructive material is no cost of any recognizable structures. (agckinase.com)
Primer3
- The graphic illustrates the primer configuration utilized during the amplification. (biosyn.com)
- Amplification speed was determined in LAMP reactions using adenovirus AdV41 synthetic DNA and Mycoplasma pneumoniae genomic DNA, their respective primer sets and SYTO 9 green fluorescent nucleic acid stain for real-time detection. (thermofisher.com)
- This enzyme extends the primer according to the nucleic acid sequence of the template (adding dNTPs or nucleotides to the end of the primer). (mantacc.com)
Genetic1
- A virus is an infectious microorganism made up of a nucleic acid segment, which may contain any type of genetic material like deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) coated inside a protein layer. (biomedcentral.com)
Molecular Biology2
- The isothermal nucleic acid amplification engineering (INAAT) is made use of in the industry of molecular biology and recombinant DNA systems for detecting and analyzing nucleic acids. (floschi.info)
- The present invention relates generally to the fields of molecular biology and nucleic acid analysis. (justia.com)
LAMP2
- The LAMP technique is the foundation of the Complex DNA Lyo Kit. (medgadget.com)
- We analyse the techniques designed to miniaturize nucleic acid amplification and bring it towards point-of-care, such as PCR, LAMP, and NEAR. (idtechex.com)
Sequence3
- Molecular technique is further segmented into nucleic acid sequence based amplification and PCR. (prnewswire.co.uk)
- In their paper, the two scientists describe an in-vitro procedure for the selective amplification of closed circular DNA using sequence-specific primers. (biosyn.com)
- We will take a closer look at the first two methods, amplification-based and sequence-based. (integra-biosciences.com)
Detect2
- The 'Cycle threshold' or Ct value is the number of amplification cycles required to detect the first signal that the virus is present. (rivm.nl)
- In this review, we have explained a few types of diagnostic techniques: Biosensor based, immunological-based, and molecular-based diagnostic techniques that are prominent methodologies to identify and detect the course of infection related to the medical viruses. (biomedcentral.com)
Fluorescent2
- However, problems exist with current techniques used to create fluorescent primers. (nih.gov)
- The technique converts a biological response into a measurable signal without the aid of fluorescent labels or other tags and tagging agents. (strategic-directions.com)
INAAT4
- The worldwide market for isothermal nucleic acid amplification technology (INAAT) is anticipated to increase as a result of major companies increasingly developing highly effective and user-friendly INAAT-based solutions. (medgadget.com)
- This useful gadget relies on the isothermal nucleic acid amplification technology concept (INAAT). (medgadget.com)
- The worldwide market for isothermal nucleic acid amplification technology (INAAT) is anticipated to expand as a result of the rising product and technological improvements for DNA amplification. (medgadget.com)
- Due to an increase in product launches by major players, the worldwide isothermal nucleic acid amplification technology (INAAT) market is anticipated to grow at a CAGR of 8.3% between 2019 and 2027. (medgadget.com)
Throughput2
- Furthermore, the miniaturization and automation of ASOF analysis leads to an exceedingly increased throughput of nucleic acid analysis. (justia.com)
- Advances in nucleic acid sample preparation have facilitated increased throughput and speed, the ability to work with smaller sample sizes, and increased effectiveness for more complex sample types. (strategic-directions.com)
Electrophoresis1
- This involves separating nucleic acid fragments by electrophoresis, staining nucleic acid fragments with embedded dyes such as ethidium bromide or SybrSafe, and then detecting with a UV light source and imaging system (Figure 2). (mantacc.com)
ELISA1
- Biochemical technique is further segmented into enzyme immunoassay, agglutination and ELISA. (prnewswire.co.uk)
20192
- For instance, in June 2019, researchers from the University of Electro-Communications, Abbott Japan Co., Ltd., and the Tokyo Institute of Technology created DNA L-TEAM, a revolutionary technique for amplifying DNA at Low-temperature Amplification. (medgadget.com)
- The international isothermal nucleic acid amplification technological innovation market grew at a CAGR of all over 10% all through 2014-2019. (floschi.info)
Fragments1
- Photo of DNA Agarose Gel: The far-left row is a DNA ladder, containing nucleic acid fragments of known sizes. (mantacc.com)
Deoxyribonucleic acid1
- In this chapter, aspects of avian mycobacteriosis, including clinical presentation, therapeutic options, zoonotic potential and diagnostic tests with promising new molecular techniques such as deoxyribonucleic acid (DNA) probes, will be covered. (ivis.org)
Assay1
- The isothermal technique allows for positive results to be available as soon as 5 min into the assay, and negative results within 13 min. (elifesciences.org)
Tests1
- Tests are usually performed using rapid immunochromatographic strips or nucleic acid amplification techniques. (wkinformation.com)
Pathogen1
- Conventional technique is further segmented into gram-stain and pathogen culturing. (prnewswire.co.uk)
Fluorescence2
- Additionally, the GEAR isothermal method can be performed in a relatively inexpensive water bath or heating block, with detection of amplification products by fluorescence, thus making it suitable for low resource settings. (nih.gov)
- Real-Time PCR Amplification Curve: The graph correlates the fluorescence signal with the number of temperature cycles. (mantacc.com)
Methods3
- In order to ensure lesser chlamydia complications, Reliable diagnostic techniques and effectives treatment methods play indispensable role in reducing the degree of severity in patients. (streetinsider.com)
- The study's primary objective is to compare conventional methods such as acid-fast bacillus (AFB) culture and microscopy with rapid diagnostic methods. (scielo.br)
- Molecular diagnostics techniques are emerging as the tools of choice for discerning both the identity of pathogens and their mechanisms of resistance, either as an alternative approach or to complement conventional methods. (integra-biosciences.com)
Specificity1
- Specificity is highest on sputum that is positive by acid-fast staining. (aafp.org)
Thermal3
- Thermal denaturation separates the two circular DNA strands that can now serve as the template for the next round of extension and amplification. (biosyn.com)
- The technique is cost effective because it is carried out at a constant temperature and does not require a thermal cycler or other expensive equipment. (thermofisher.com)
- The ID NOW system is a point-of-care (POC) device that uses an isothermal nucleic acid amplification technique to allow for nucleic acid amplification without thermal cyclers and allows for results to be obtained quickly. (elifesciences.org)
Approaches2
- A Feature Paper should be a substantial original Article that involves several techniques or approaches, provides an outlook for future research directions and describes possible research applications. (mdpi.com)
- Established nucleic acid detection approaches can be similarly sensitive, but SHERLOCK gave more consistent results. (nih.gov)
Diagnostic1
- Market driving factors responsible for the growth of infectious disease diagnostic market include growing incidences of infectious disease, emphasis on advanced molecular techniques in underdeveloped regions. (prnewswire.co.uk)
Bacilli3
- Acid-fast bacilli (AFB) smear and culture using sputum obtained from the patient. (medscape.com)
- His acid-fast bacilli C/S reports during follow-up showed growth of Mycobacterium Avium complex (MAC). (japt.in)
- Bronchoscopy was done and bronchoalveolar lavage (BAL) samples sent for acid-fast bacilli (AFB) smear was negative, Gene X-pert MTB was not detected, but cytology showed granuloma lesions. (japt.in)
Diagnostics2
- Based on techniques, the infectious diagnostics disease market is segmented into conventional techniques, biochemical techniques and molecular techniques. (prnewswire.co.uk)
- We can now effectively and readily make sensors for any nucleic acid, which is incredibly powerful when you think of diagnostics and research applications," Collins says. (nih.gov)
Specific1
- NEAA reactions also present some drawbacks, (e.g., severe non-specific amplification and strong background signals. (nih.gov)
Primers1
- Chen and Ruffer used a circular plasmid of 1990 base pairs (bp), and two 5' phosphorylated primers,16 and 17 nucleotides (nt) long, for the generation and amplification of closed circular DNA. (biosyn.com)
Laboratory1
- A summary of the laboratory techniques for GBS identification is depicted in Ref 18. (wikipedia.org)
Routine1
- Nucleic acid testing (NAT) in routine HIV testing programs can increase the detection of infected individuals, but the most effective implementation of NAT remains unclear. (nih.gov)
Researchers2
- The researchers took advantage of this property to design a CRISPR-based nucleic acid detection platform. (nih.gov)
- Researchers suggest that mobiLab may be an effective-and cheaper-screening technique and are hopeful more healthcare facilities will offer this testing method to reach a wider population. (doctorshealthpress.com)
Sialic1
- As other virulent bacteria, GBS harbours an important number of virulence factors, The most important are the capsular polysaccharide (rich in sialic acid), and a pore-forming toxin, β-haemolysin. (wikipedia.org)
