A Real-Time Fluorescence Polymerase Chain Reaction Assay for the Identification of Yersinia pestis Using a Field-Deployable Thermocycler

Abstract

Real-time fluorescence polymerase chain reaction is a microbial identification method that can provide rapid and accurate results using a field-deployable thermocycler, the RAPID ("ruggedized" advanced pathogen identification device). A Yersinia pestis-specific TaqMan assay required approximately 75 minutes and achieved a sensitivity of 100 fg of Y. pestis genomic DNA (20 genome equivalents). Specificity testing against a genomic DNA cross-reaction panel comprised of 22 bacterial species encountered in the respiratory tract resulted in no false positives. No cross-reaction occurred with human genomic DNA.

Primates and the Ecology of their Infectious Diseases: How will Anthropogenic Change Affect Host-Parasite Interactions?

“…55 In addition, thermocyclers with optical capabilities, which are necessary for real-time PCR applications, are shrinking in size and weight, while lyophilized reagents have become available that are stable at room temperature for extended periods, making it possible to transport these new diagnostic tools to remote field sites. 56 Although such technologies are primarily being developed for military and agricultural applications, only minor adaptations would be required for the rapid diagnosis of primate infectious disease in field settings. It should soon be possible to screen large numbers of primates for a series of parasites in "real time" at remote field sites, and to generate accurate and precise measurements of seasonal variation in infectious disease prevalence and intensity.…”

Real-Time PCR Detection of Campylobacter Spp. In Free-Ranging Mountain Gorillas (Gorilla Beringei Beringei)

“…Molecular diagnostics have long been used on noninvasively collected samples from humans to diagnose infection with a variety of pathogens, including many potentially pathogenic to great apes (Lina et al, 1996;Houng et al, 1997). Wider implementation of molecular diagnostic techniques could facilitate free-ranging great ape disease research, especially using noninvasive samples coupled with durable, portable technology capable of providing rapid results in the field (McAvin et al, 2003;Tomlinson et al, 2005).…”

Comparison of five commercial DNA extraction kits for the recovery of Yersinia pestis DNA from bacterial suspensions and spiked environmental samples

“…Consequently, Y. pestis has been classified as a Category-A Critical Biological Agent by the Centers for Disease Control and Prevention (CDC) (Rotz et al 2002). The virulence of Y. pestis, as well as its potential as an agent of bioterrorism, has led to the development and validation of several real-time PCR assays for the rapid detection of Y. pestis in a variety of specimen types (Higgins et al 1998;Loiez et al 2003;McAvin et al 2003;Tomaso et al 2003;Woron et al 2006;Stewart et al 2008).…”