Polymerase chain reaction product detection

Schwartz, H. E., Ulfelder, K., Sunzeri, F. J., Busch, M. R, and Brownlee, R. G., Analysis of DNA restriction fragments and polymerase chain reaction products towards detection of the AIDS (HIV-1) virus in blood, /. Chromatogr., 559, 267, 1991. 

Hurst, G. Doktycz, M. Vass, A. Buchanan, M. Detection of bacterial DNA polymerase chain reaction products by matrix assisted laser desorption/ionization mass spectrometry. Rapid. Commun. Mass Spectrom. 1996,10,377-382. 

Sanchez-Rangel, D., Sanjuan-Badillo, A., and Plasencia, J. (2005). Fumonstn production by Fusarium verticillioides strains isolated from maize in Mexico and development of a polymerase chain reaction to detect potential toxigenic strains in grains. Agric. Food Chem. 53, 8565-8571. 

McWhorter S, Soper SA. Conductivity detection of polymerase chain reaction products separated by micro-reversed-phase liquid chromatography. Journal of Chromatography A 883,1-9, 2000. 

Analysis of polymerase chain reaction-product by capillary electrophoresis with laser-induced fluorescence detection and its application to the diagnosis of medium-chain acyl-coenzyme A dehydrogenase deficiency. 

M. Galloway and S.A. Soper, Contact conductivity detection of polymerase chain reaction products analyzed by reverse-phase ion pair microcapillary electrochromatography, Electrophoresis, 23 (2002) 3760-3768. M. Masar, M. Dankova, E. Olvecka, A. Stachurova, D. Kaniansky and B. Stanislawski, Determination of free sulfite in wine by zone electrophoresis with isotachophoresis sample pretreatment on a column-coupling chip, J. Chromatogr. A, 1026 (2004) 31-39. 

Holland, P.M. Abramson, R.D. Watson, R. Gelfand, D.H. Detection of specific polymerase chain reaction product by utilizing the 5 to 3 exonuclease activity of Thermus aquaticus. Proc. Natl. Acad. Sci. 1991, 88, 7276-7280. 

Suzuki Y, Orita M, Shiraishi M, Hayashi K, Sekiya T. Detection of ras gene mutations in human lung cancers by single-strand conformation polymorphism analysis of polymerase chain reaction products. Oncogene 1990 5 1037-43. 

Muhammad WT, Tabb DL, Fox KF, and Fox A (2003) Automated discrimination of polymerase chain reaction products with closely related sequences by software-based detection of characteristic peaks in product ion spectra. Rapid Communications in Mass Spectrometry 16 1-8. 

Walters JJ, Fox KF, and Fox A (2002) Mass spectrometry and tandem mass spectrometry, alone or after liquid chromatography, for analysis of polymerase chain reaction products in the detection of genomic variants. Journal of Chromatography B 782 57-66. 

Vlieger AM, Medenblik AM, van Gijlswijk RP, Tanke HJ, van der Ploeg M, Gratama JW, and Raap AK (1992) Quantitation of polymerase chain reaction products by hybridization-based assays with fluorescent, colorimetric, or chemiluminescent detection. Analytical Biochemistry 205(1) 1-7. 

Honeycutt, R., Sobral, B. W and McClelland, M. (1997) Polymerase chain reaction (PCR) detection and quantification using a short PCR product and a synthetic internal positive control. Anal. Biochem. 248, 303-306. 

Nurmi J, Ylikoski A, Soukka T et al (2000) A new label technology for the detection of specific polymerase chain reaction products in a closed tube. Nucleic Acids Res 28 e28... 

Lopez E., C. Chypre, B. Alpha, G. Mathis. Europium tris(bipyridyne) cryptate label for time-resolved luminescence detection of polymerase chain reaction products fixed on a solid support, Clin. Chem., 39, 196-201 (1993). 

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. 

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