Quantitative, polymerase chain reaction

Quantitative PGR (Q-PGR or real-time PGR) detects specific DNA amplification products being formed by the polymerases. As with classic spectrophotometric methods, the signal arising from the bound fluorescently-labeled nucleotide probe increases in direct proportion to the amount of PGR product formed during reaction. Because the method is real-time, Q-PGR eliminates the need for post-PGR processing thus 

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Quantitative polymerase chain reaction, also called real-time RT-PCR or QPCR, is a method which employs insertion of a signal, such as fluorescence or enzyme activity, into PCR products generated by RT-PCR to determine the amount of messenger RNA (mRNA) in a tissue accurately. 

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... 

Nakamura, T., Sakaeda, T., Ohmoto, N., Tamura, T., Aoyama, N. et al., Real-time quantitative polymerase chain reaction for MDR1, MRP1, MRP2, and CYP3A-mRNA levels in Caco-2 cell lines, human duodenal enterocytes, normal colorectal tissues, and colorectal adenocarcinomas, Drug Metab. Dispos. 2002, 30, 4—6. 

Quantitative Polymerase Chain Reaction (QPCR) This method can determine the amount of DNA or RNA in a sample. Using the QPCR method, DNA can be amplified many times, allowing minute quantities to be assessed. 

Cruz-Perez, P., Buttner, M. P., and Stetzenbach, L. D. (2001). Specific detection of Stachybotrys chartarum in pure cultures using quantitative polymerase chain reaction. Mol. Cell. Probes 15,129-138. 

Bortolin, S. Christopoulos, T. K. Verhaegen, M. Quantitative polymerase chain reaction using a recombinant DNA internal standard and time-resolved fluorometry. Anal. Chem. 1996, 68(5), 834-840. 

Becker, K. Pan, D. Whitley, . B. Real-time quantitative polymerase chain reaction to assess gene transfer. Hum. Gene. Ther. 1999, 70(15), 2559-2566. 

Fig. 8.2.2 Principle of genomic quantification by multiplex ligation-dependent probe amplification (MLPA). Synthetic oligonuleotides are designed to bind to exon 1 (oligo 1a and 1b) and 2 (2a and 2b). After specific hybridisation, the oligos are ligated. The ligation products undergo quantitative polymerase chain reaction using universal primers X and Y...

Van den Heuvel JP, Clark GC, Thompson CL, et al. 1993. CYP1A1 mRNA levels as a human exposure biomarker use of quantitative polymerase chain reaction to measure CYP1A1 expression in human peripheral blood lymphocytes. Carcinogenesis 14 2003-2006. 

Bieche I, Olivi M, Champeme MH, Vidaud D, Lidereau R, Vidaud M. Novel approach to quantitative polymerase chain reaction using real-time detection application to the detection of gene amplification in breast cancer. Int J Cancer 1998 78(5) 661-666. 

Li, B., Sehajpal, P. K., Khanna, A., Vlassara, H., Cerami, A. et al. (1991). Differential regulation of transforming growth factor beta and interleukin 2 genes in human T cells Demonstration by usage of novel competitor DNA constructs in the quantitative polymerase chain reaction./. Exp. Med. 174, 1259-1262. 

Biodistribution analysis is conducted at the molecular level. The current gold standard is a quantitative polymerase chain reaction (Q-PCR) assay that detects the number of vector copies per microgram of genomic DNA. Biological fluids and tissue samples are carefully harvested (to avoid crosscontamination) from control and gene therapy product-injected animals at... 

For HAM/TSP disease development, HTLV-I proviral load is an important factor. Recently, a real time quantitative polymerase chain reaction (PCR) assay (TaqMan) was developed... 

Biodistribution studies are conducted to evaluate the dissemination and persistence of nucleic acid/DNA and viral vector-based vaccines. Vector persistence should be examined at more than one time point up to at least 21 days or more after the last injection 22 unpublished data) and the biodistribution and persistence of a plasmid is usually dependent upon the route of administration.20 These studies can be combined with a general toxicology study or may be conducted separately. In most cases, the DNA is extracted from the selected tissues and assessed by quantitative Polymerase Chain Reaction (qPCR).1516... 

Feldman AM, Ray PE, Bristow MR. Expression of a-subunits of G proteins in failing human heart a reappraisal utilizing quantitative polymerase chain reaction. J Mol Cell Cardiol 1991 23 1355-1358. 

Ferreira-Gonzalez A, Fisher RA, Weymouth LA, Langley MR, Wolfe L, WiUdnson DS, et al. Clinical utility of a quantitative polymerase chain reaction for diagnosis of cytomegalovirus disease in solid organ transplant patients. Transplantation 1999 68 991-6. 

Roberts TC, Brennan DC, Buller RS, Gaudreault-Keener M, Schnitzler MA, Sternahell KA, et al. Quantitative polymerase chain reaction to predict occurrence of symptomatic cytomegalovirus infection and assess response to ganciclovir therapy in renal transplant recipients. J Infect Dis 1998 178 626-35. 

Ma XJ, Patel R, Wang X, et al. Molecular classification of human cancers using a 92-gene real-time quantitative polymerase chain reaction assay. Arch Pathol Lab Med. 2006 130 465-473. 

G8. Guller, U., Zajac, P., Schnider, A., Bosch, B., Vorburger, S., Zuber, M., et al. Disseminated single tumor cells as detected by real-time quantitative polymerase chain reaction represent a prognostic factor in patients undergoing surgery for colorectal cancer. Ann. Surg. 236, 768-776 (2002). 

Quantitative polymerase chain reaction does not improve preoperative prostate cancer staging a clinicopathological molecular analysis of 121 patients. / Urol, 156, 1560-1566. 

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