Polymerase chain reaction , real-time

Cultures are subject to time delay. New methodology using real-time polymerase chain reaction (PCR) shortens the reporting time. 

Fortin, N. Y. Mulchandani, A. Chen, W. Use of real-time polymerase chain reaction and molecular beacons for the detection of Escherichia coli 0157 H7. Anal. Biochem. 2001, 289, 281-288. 

Kuboniwa, M. Amano, A. Kimura, K. R. Sekine, S. Kato, S. Yamamoto, Y. Okahashi, N. Iida, T. Shizukuishi, S. Quantitative detection of periodontal pathogens using real-time polymerase chain reaction with TaqMan probes. Oral Microbiol. Immunol. 2004,19,168-176. 

Badosa E, Trias R, Pares D, Pla M and Montesinos E. 2008. Microbiological quality of fresh fruit and vegetable products in Catalonia (Spain) using normalised plate-counting methods and real time polymerase chain reaction (QPCR). J Sci Food Agric 88(4) 605-611. 

J.R. Uhl, C.A. Bell, L.M. Sloan, M.J. Espy, T.F. Smith, J.E. Rosenblatt and F.R. Cockerill, Application of rapid-cycle real-time polymerase chain reaction for the detection of microbial pathogens the Mayo-Roche rapid anthrax test, Mayo Clin. Proc., 77 (2002) 673-680. 

Nelson SM, Ferguson LR, Denny WA (2005) Demonstration by real time polymerase chain reaction that cellular DNA alkylation by novel aminoindoline compounds affects expression of the protooncogene c-myc. Chem Res Toxicol 18(2) 239-248... 

Figure 2 Cytokine gene expression in immunopotentiating reconstituted influenza virosomes (IRIV) stimulated peripheral blood mononuclear cells (PBMC). PBMC were cultured in the presence or absence of IRIV. On days 1 and 2, culture cells were harvested and total cellular RNA was extracted and reverse transcribed. The cDNAs thus obtained were tested in real time polymerase chain reaction assays in the presence of primers specific for the indicated cytokine genes. Source From Refs. 6 and 9.

Fig. 9.4 Establishment of the quantitative methylation-specific polymerase chain reaction (MSP) analytical procedure. The ABI PRISM 7900 HT Sequence Detector system was used to perform real-time polymerase chain reaction (PCR) using MSP primers and bisulfite-modified template DNA. Upper panels Setting up the conditions to obtain the standard curves with 50% (A) or 25% (B) sequential dilution of the template. Lower panels The amplification curves on the left represent P-actin, unmethylated, and methylated MSP products, respectively, for reelin (RELN) (C). Amplification curves were compared at the set threshold before 40 cycles. Amplification curves from various samples are shown in the lower panel right (D)...

Real-time polymerase chain reaction (RT-PCR), microarray... 

FIGURE3.10 UCP1 mRNA expression levels in epididymal WAT (Okada et al., 2011). Expression of UCP1 mRNA was estimated by quantitative real-time polymerase chain reaction. Relative values were presented as the ratio of UCP1 mRNA to GAPDH mRNA. 

It should be possible to develop either biochemical- or molecular-based tests for resistance in flatworms, as has been successfully used to detect insecticide resistance in insects. However, the necessary information is not available on the mechanism of resistance to be able to design tests of this type for use with trematodes or cestodes. When developed they will hopefully be based on real-time polymerase chain reaction (PCR) or pyrosequencing. 

Sims PW, Vasser M, Wong WL, Williams PM, Meng YG. Immunopolymerase chain reaction using real-time polymerase chain reaction for detection. Anal Biochem 2000 281(2) 230-232. 

Assimilable organic carbon (AOC) was determined in water using flow-cytometric enumeration and a natural microbial consortium as inoculum.134 Two bacterial species were used for the measurement of AOC in water, based on their respective 16S rDNA sequences. The AOC content in 41 water samples was determined with these two sets by quantitative real-time polymerase chain reaction (qRT-PCR).135... 

Chang TK, Chen J, Pillay V, Ho JY, Bandiera SM. Real-time polymerase chain reaction analysis of CYP1B1 gene expression in human liver. Toxicol Sci 2003 71 11-19. 

In another example, real-time polymerase chain reaction (RT-PCR) was used to determine expression levels of the 48 known human ABC-type transporters in the NCI-60 cell lines, and these levels were correlated with sensitivity to 1429 candidate anticancer drugs (149, 150). Patterns of expression correlated moderately well with tissue of origin and were independent of sequence homology among... 

Beller H. R., Kane S. R., Legler T. C., and Alvarez P. J. J. (2002) A real-time polymerase chain reaction method for monitoring anaerobic, hydrocarbon-degrading bacteria based on a catabolic gene. Environ. Sci. Technol. 36(18), 3977-3984. 

Lekanne Deprez, R.H. Fijnvandraat, A.C. Ruijter, J.M. Moorman, A.F.M. Sensitivity and accuracy of quantitative real-time polymerase chain reaction using SYBR green... 

Real-Time Polymerase Chain Reaction Methods... 

Helps et al. (2004) slaughtered two female cattle, followed by one male, and then four female cattle and collected swab samples from the split vertebral-column surfaces. Real-time polymerase chain reaction protocols were followed to determine the extent to which tissue from the male carcass accumulated in the splitting saw and was disseminated to subsequent female carcasses. Under simulated abattoir conditions (i.e., washing the saw for 5 s between carcasses and washing the carcasses before collecting samples), these researchers reported that 0.01% of the tissue recovered from the split vertebral-column surface of the final female carcass in the sequence was from the male carcass and that 10% of the tissue remaining in the housing of the saw was from the male carcass. It was concluded from that study that "should a BSE-positive carcass be... 

Pryor RJ, Wittwer GT. Real-time polymerase chain reaction and melting curve analysis. Methods Mol Biol. 2006 336 19-32. 

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