5′ Nuclease PCR Assay To Detect Yersinia pestis

Rapid identification of Yersinia enterocolitica in blood by the 5' nuclease PCR assay.

Yersinia enterocolitica accounts for 50% of the clinical sepsis episodes caused by the transfusion of contaminated red blood cells. A 5' nuclease TaqMan PCR assay was developed to detect Y. enterocolitica in blood. Primers and a probe based on the nucleotide sequence of the 16S rRNA gene from Y. enterocolitica were designed. Whole-blood samples were spiked with various numbers of Y. enterocolit...

Development of a fluorogenic 5' nuclease PCR Assay for the detection of pathogenic Yersinia enterocolitica

In this report, we describe the development and evaluation of a 5' nuclease PCR assay for the detection of pathogenic Yersinia enterocolitica (YE). The assay targets the chromosomally encoded invasion gene ail. Three different primer/probe sets (TM1, TM2, and TM3) amplifying different, yet overlapping, regions of ail were examined for their specificity and sensitivity. The TM1 set displayed the...

Yersinia pestis Real Time PCR Kit

Modified method to detect PCR products by 5' nuclease activity and an asymmetric fluorogenic probe set.

PCR products are detected most commonly using postamplification agarose gel electrophoresis and/or DNA hybridization. For reducing time in detection and confirmation, a number of investigators have reported methods to simplify or even eliminate the need to perform postamplification assays. For example, molybodate (2) or some intercalating agents (3) have been used to measure the degree of DNA a...

Temperature-modulated immunogenicity to Yersinia pestis from Yersinia enterocolitica O3.

The ability of Yersinia enterocolitica O3, grown at 25 degrees C, to promote cross-immunity to Y. pestis was lost after repeated subcultures at 37 degrees C, which selected for bacterial populations having lower in vivo survival. Subculturing Y. enterocolitica O3 from 37 to 25 degrees C restored the cross-immunogenicity although the in vivo survival remained low.