Polymerase chain reaction infections

Spann, W., Pachmann, K., Zabnienska, H., Pielmeier, A., and Emmerich, B. (1991) In-situ amplification of single copy gene segments in individual cells by polymerase chain reaction. Infection 19, 242-244. 

The plaque assay is desirable because it is very sensitive and only detects infectious viral particles. However, there are viral agents which cannot be supported by cell lines. In these cases other methods must be used. The polymerase chain reaction (PGR), which amplifies DNA or RNA from viral agents, can be used to detect the presence and quantity of viral agents. The amount of RNA or DNA target in the initial sample can be determined by competitive PGR where the quantity of amplified product is compared to a control PGR product where the initial amount of target is known. Quantification is also possible by an end-point dilution method similar to that used to determine a tissue culture infections dose. PGR methods can be very sensitive however. 

Lok AS, McMahon BJ (2007) Chronic hepatitis B. Hepatology 45(2) 507-539 Manesis EK, Papatheodoridis GV, Sevastianos V, Cholongitas E, Papaioannou C, Hadziyannis SJ (2003) Significance of hepatitis B viremia levels determined by a quantitative polymerase chain reaction assay in patients with hepatitis B e antigen-negative chronic hepatitis B virus infection. Am J Gastroenterol 98(10) 2261-2267... 

Bagasra O, Lavi E, Bobroski L, Khafili K, Pestaner JP, Tawadros R, Pomerantz RJ (1996) CeUular reservoirs of HIV-1 in the central nervous system of infected individuals identification by the combination of in situ polymerase chain reaction and immunohistochemistry. Aids 10(6) 573-585... 

Hepatitis D infection requires the presence of HBV for HDV viral replication. Measuring HDV RNA levels in the serum by polymerase chain reaction (PCR) confirms the presence of... 

Confirmation of positive ELISA for HIV infection (more specific test for anti-HIV antibodies) Polymerase chain reaction (PCR) ... 

Armed with this new tool, Schena et al. (1996) created a microarray of 1,046 human cDNAs of unknown sequence. They were derived from human peripheral blood lymphocyfes fransformed wifh Epsfein-Barr virus. Suitably sized inserts [>600 base pairs (bp)] were cloned into a lambda vector, subsequently infected into an Escherichia coli strain, and finally amplified by polymerase chain reaction (PCR) using 5 -amino-modified primers. The resulting 5 -amino-modified cDNA amplicons were then arrayed onto sily-lated microscope slides. Next, the expression levels in human Jurkat cells undergoing heat shock or phorbol ester induction were examined. 

Beck, J. J., and Barnett, C. J. (2003). In "Detection of Fusarium species infecting com using the polymerase chain reaction", p. 23U. S. Patent Application 200330113722, June 19, 2003. 

E. Therapeutic response Efficacy of Infergen therapy was determined by measurement of serum alanine aminotransferase (ALT) concentrations at the end of therapy (24 weeks) and following 24 weeks of observation after the end of treatment of adults with chronic HCV infection. Serum HCV RNA was also assessed using a quantitative reverse transcriptase polymerase chain reaction (RT-PCR). At the end of 24 weeks of treatment, ALT normalization was observed in 39% of patients on Infergen and in 35% of patients on interferon alfa-2b Intron A). Only 17% of patients in each group... 

Bettinger, D., Mougin, C, and Lab, M (1994) Rapid detection of cytomegalo-virus-infection by in situ polymerase chain reaction on MRC5 cells inoculated with blood specimens J Virol Methods 49, 59-66... 

A number of phagemid display systems that fuse Ab chains to either full-length or N-terminally deleted gp3 fusions have been described, along with the polymerase chain reaction (PCR) primer sets, restriction enzymes, and host strains required for the display of scFvs and Fabs (3—7). Some systems are commercially available. There is an important difference in whether the Ab-gp3 fusions are with the whole of the protein or with just the C-terminal domain. Fusions to the whole of gp3 require that the expression of the fusion is suppressed until after it is infected with the helper phage because of the resistance... 

What are the major advantages of the polymerase chain reaction (PCR) method for amplifying defined segments of DNA as opposed to the use of conventional cloning methods How might the PCR method be used to test for infection with the AIDS virus and... 

Hamburger, J., He, N., Xin, X.Y., Ramzy, R.M., Jourdane, J. and Ruppel, A. (1998a) A polymerase chain reaction assay for detecting snails infected with bilharzia parasites (Schistosoma mansoni) from very early prepatency. American Journal of Tropical Medicine and Hygiene 59, 872-876. 

Stefanic, S., Shaikenov, B.S., Deplazes, P., Dinkel, A., Torgerson, P.R. and Mathis, A. (2004) Polymerase chain reaction for detection of patent infections of Echinococcus granulosus ( sheep strain ) in naturally infected dogs. Parasitology Research 92, 347-351. 

Viral detection assays based on infectivity suffer from significant variability, which necessitates the use of statistical evaluation. Polymerase chain reaction-based assays are currently being developed and validated for viral clearance. With PCR assays, there is a potential to distinguish between inactivation and physical removal, perform mass balance studies, evaluate more than one vims at a time for a given process step, reduce the time for completing clearance studies, and accurately quantitate the amount of vims bound to such surfaces as chromatography resins. Table 5 compares the assay precision between an infectivity assay and a quantitative PCR assay. 

Fedricks DN, Reiman DA. Application of polymerase chain reaction to the diagnosis of infectious diseases. Clin Infect Dis 1999 29 475-488. 

Wesselingh, S. L., Takahashi, K., Glass,J. D., McArthur,J. C., Griffin,J. W., and Griffin, D. E. (1997). Cellular localization of tumor necrosis factor mRNA in neurological tissue from HIV-infected patients by combined reverse transcriptase/polymerase chain reaction in situ hybridization and immunohistochemistry. J. Neuroimmunol. 74, 1—8. 

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. 

Schochetman, G. and Sninsky, J.J. (1992). Direct detection of human immunodeficiency virus infection using polymerase chain reaction. In AIDS Testing Methodology and Management Issues. G.Schochetman and J.R.George, eds. (New York Springer-Verlag), pp. 90 110. 

A clinical suspicion of an Acanthamoeba infection is the critical first therapeutic step. Acanthamoeba can be diagnosed by eye smears, culture, tissue biopsy, polymerase chain reaction, and confocal microscopy. 

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