PCR test

Diagnostic tests for amebiasis include stool for ova, antigen detection, or polymerase chain reaction (PCR) testing. 

Combining both heating and nonheating protocols employed in a sequential order were evaluated, but without any advantage (Fig. 3.4). RT-PCR was performed by standard methods, RNA extracted from fresh MDA cells and human tissue of breast cancer with known tested genes was used as positive control, and pure water was used to replace template (cDNA) as negative control for every experiment of PCR. To assure the accuracy of PCR tests, all reactions were performed in triplicate. 

Note A simplified protocol of DNA extraction from FFPE tissue may be modified by using heat treatment alone without subsequent purification steps as a one-step protocol boiling the FFPE tissue sections in alkalin solution (0.1 M NaOH) for 20min, cooled down for 15min, and 0.2pF of the retrieval solution is aspirated for PCR test and the remaining sample stored at 4°C for further use. 

PCR Testing for Sickle Cell Disease with ASO Probes... 

Brown WT, Houck GE, Jeziorowska A, et al. Rapid fragile X carrier screening and prenatal diagnosis using a non-radioactive PCR test. JAMA 270 1569-1575,1993. 

PCR testing, etc. GLPs with the exception of. .. of the toxicology study Methodology/standardization of assays evolving (level of sensitivity improving) Pre-GMP test material Laboratory procedures draft SOPs QA review of the final report only... 

Figure 11.6. A typical restriction length polymorphism-polymerase chain reaction (RFLP-PCR) test for the CYP2C19 2 mutation. Complementary primers that are specific for CYP2C19 (heavy arrows) are used to amplify a 312-bp fragment. The DNA is digested with a restriction enzyme Smal and electrophoresced on an agarose gel. The wild-type DNA is cut by Smal into two pieces of 109 bp and 212 bp, whereas the homozygous mutant allele is not cut. Individuals heterozygous for the two alleles show all three bands.

Polymerase chain reaction (PCR) techniques can be used to diagnose meningitis caused by M meningitidis, S. pneumoniae, and H. influenzaetype b (Hib). PCR is considered to be highly sensitive and specific. PCR testing of the CSF is the preferred method of diagnosing most viral meningitis infections. 

A positive anti-HCV test should be confirmed by means of the RT-PCR test or quantitative hybridization test, e. g. branched-DNA hybridization, amplicor technique. With the help of these procedures, it is possible to detect HCV RNA within 1 week after infection. (104, 105, 108,... 

The detection of viraemia can only be carried out by determining the HCV RNA in the PCR test after prior reverse transcription (RT) (K.B. Mullis, 1985). This also works in the incubation phase with patients who are still anti-HCV-negative with normal transaminase values as well as with serum that is still anti-HCV-negative in the acute phase. Thus the following indications are given for the PCR method ... 

Check again for the absence of EMCV DNA by repeating the PCR testing. If negative PCR results are seen, proceed to check the functional quality of the template. 

Enzyme-linked immunoabsorbent assays (ELISAs) for VHP infections can be performed on samples inactivated by treatment with P-propiolactone. Reverse transcriptase polymerase chain reaction (RT-PCR) tests on samples following RNA extraction using chloroform and methanol can detect most of the VHP agents rapidly. RT-PCR is particularly useful when isolation of the virus is difficult or impractical, and was effective in detecting the agent causing HPS months before it was isolated in culture (48). 

Bourlet T, Caro V, MmjoUe S, Jusselin I, Pozzetto B, Crainic R, et al. New PCR test that recognizes all human prototypes of enterovirus application for clinical diagnosis. J Clin Microbiol 2003 41 ... 

Kleiber J, Walter T, Haber hausen G, Tsang S, Babiel R, Rosenstraus M. Performance characteristics of a quantitative, homogeneous TaqMan RT-PCR test for HCV RNA. J Mol Diagn 2000 2 158-66. 

In the absence of an unambiguous history of ricin exposure, the preferred diagnostic method is specific immunoassay of ricin in serum, respiratory secretions, or other clinical samples associated with poisoning. Most of the methods described for ricin detection are experimental or are under development. The CDC and the Federal Laboratory Response Network have the capability to detect ricin in environmental specimens using validated polymerase chain reaction (PCR) tests and time-resolved immunofluorescence assays, with cell-based bioassays to confirm ricin activity. The U.S. Department of Defense has produced experimental field immunoassays, but commercial distribution of field test kits currently is limited. 

PCR testing of the CSF is the preferred method of diagnosing most viral meningitis infections. 

Newer diagnostic tests such as monoclonal antibody or DNA probe techniques, as well as PCR tests that can detect small amounts of trichomonal DNA, have been developed. These tests are highly sensitive and specific for detecting infection in both urogenital spec-... 

The research formats presently used in clinical reference laboratories employ complex interpretation schemes, though they have been streamlined to be as decisive as is practical. Compatible with traditional interpretation, the first decision to be made is whether the test is valid through examination of the control results. PCR tests may include controls to test for sample preparation, amplification, and the detection of the amplified DNA. Additionally, for each patient specimen, an internal amplification control (e.g., for normal and mutant genes) may be included. Table 6 lists one interpretation scheme for an HIV assay wherein one primer pair and probe are used in the test and the decision is based on replicate testing. 

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