The objective of the current study was to evaluate the reproducibility of a reverse transcriptase PCR (RT-PCR)-based technique to differentiate viable and dead Salmonella cells in raw and sterilized milk. The microorganism was initially inoculated into the milk samples followed by incubating at 37°C for 4 h prior to inactivation by heat at 80°C for 10 min. The treated and non-treated samples we...

backgrounds and aims: coxsakievirus b3 (cvb3), one of the six coxsakievirus b serotypes, is a member of the enterovirus genus within the picornaviridae family. cvb3 is an important pathogen of viral myocarditis, which accounts for more than 50% of viral myocarditis cases. the genome of cvb3, like that of other entroviruses, is a single-stranded, sense, polyadenylated rna molecule with 7400 nucl...

Reverse transcription PCR (RT-PCR) represents a sensitive and powerful tool for analyzing RNA. While it has tremendous potential for quantitative applications, a comprehensive knowledge of its technical aspects is required. Successful quantitative RT-PCR involves correction for experimental variations in individual RT and PCR efficiencies. This review addresses the mathematics of RT-PCR, choice...

Avian influenza virus (AIV) infection is a major cause of bird or human mortality and morbidity, therefore the rapid identification of the virus is of important clinical and epidemiological implication. Methods: A multiplex Reverse Transcriptase PCR (RT-PCR) was optimized for the detection of influenza A virus and the H5 and H9 subtypes. The influenza type A specific primers were directed to t...

In this paper we determine the limits and accuracy of quantitative reverse transcription (RT)-PCR using a modification of the original protocol. The quantification of mRNA with this procedure requires a preliminary estimation of the target molecule (TM) concentration, established from experiments with an internal control molecule (ICM). A definitive quantification is then attained from serial d...

The present studies demonstrate a theoretical and practical framework for the accurate quantitation of gene expression in RNA extracted from microscopic tissue samples. The approaches are developed around competitive RT-PCR techniques. Assay performance has been examined and validated at both the RT and PCR steps. Our analysis of RT transcription efficiency for a number of native and competitor...