Detection and Identification of <i>Yersinia pestis</i> by Polymerase Chain Reaction (PCR) Using Multiplex Primers

Tsukano, Hiroko; Itoh, Kiyohiko; Suzuki, Sosuke; Watanabe, Haruo

doi:10.1111/j.1348-0421.1996.tb01140.x

Cited by 38 publications

(34 citation statements)

References 13 publications

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“…Extremely sensitive methods that utilize oligonucleotide targets to unique determinants are now used in many public health centers routinely to screen clinical specimens received from the field. Paramount in this effort was the use of polymerase chain reaction (PCR) for amplification of DNA sequences capable of interacting with probes prepared from pla, caf1 or both (Campbell et al, 1993;Hinnebusch and Schwan, 1993;Norkina et al, 1993;Tsukano et al, 1996). Another such method that does not rely on unique structural genes is the procedure of Trebesius et al (1998), which depends upon amplification of a 23S rRNA sequence specific to Y. pestis.…”

Section: Identificationmentioning

confidence: 98%

Yersinia pestis and Bubonic Plague

Brubaker¹

2006

The Prokaryotes

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Section: Identificationmentioning

confidence: 98%

Yersinia pestis and Bubonic Plague

Brubaker¹

2006

The Prokaryotes

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“…Dies trifft auch für den Nachweis von Y. pestis-spezifischen Genabschnitten (wie z. B. des Fra1-Kapselantigen-Gens,des V-AntigenGens,des Plasminogenaktivator-Gens) in Verdachtsproben zu [35,36,37,38,39,40,41]. Zur schnelleren Auswertbarkeit der Assays können auch Lightcycler-, Smartcycler-oder TaqMan-Technologien zum Einsatz kommen.…”

Section: Diagnoseunclassified

Yersinia pestis

Rakin¹

2003

Bundesgesundheitsbl - Gesundheitsforsch - Gesundheitsschutz

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A. Rakin Yersinia pestis. A threat to mankindAbstract Plague is a highly contagious bacterial vector-borne disease transmitted by rodent fleas (vectors). Yersinia pestis,the causative plague agent, infects human (target)s only under special conditions, resulting in severe forms of disease with high lethality.Plague cannot be eradicated, but it can be put under surveillance in the endemic loci of infection, which are distributed throughout the world. Plague is supposed to be one of the most probable candidates for intentional release in bioterrorist attacks.Although it is impossible to exclude initial casualties, we can greatly decrease them by efficient early detection of the bacteriological agent and immediate response.

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“…Conventional PCR-gel electrophoresis method has been developed for detecting Y. pestis in fleas and other specimens [12],[13],[14]. A handful of real-time quantitative PCR assays in various formats were also established for detecting and identifying Y. pestis [15],[16],[17],[18],[19],[20],[21],[22] [23].…”

Section: Introductionmentioning

confidence: 99%

Ambient Stable Quantitative PCR Reagents for the Detection of Yersinia pestis

Shi

Zhou

et al. 2010

PLoS Negl Trop Dis

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BackgroundAlthough assays for detecting Yersinia pestis using TaqMan probe-based real-time PCR have been developed for years, little is reported on room-temperature-stable PCR reagents, which will be invaluable for field epidemic surveillance, immediate response to public health emergencies, counter-bioterrorism investigation, etc. In this work, a set of real-time PCR reagents for rapid detection of Y. pestis was developed with extraordinary stability at 37°C.Methods/Principal FindingsTaqMan-based real-time PCR assays were developed using the primers and probes targeting the 3a sequence in the chromosome and the F1 antigen gene caf1 in the plasmid pMT1of Y. pestis, respectively. Then, carbohydrate mixtures were added to the PCR reagents, which were later vacuum-dried for stability evaluation. The vacuum-dried reagents were stable at 37°C for at least 49 days for a lower concentration of template DNA (10 copies/µl), and up to 79 days for higher concentrations (≥102 copies/µl). The reagents were used subsequently to detect soil samples spiked with Y. pestis vaccine strain EV76, and 5×104 CFU per gram of soil could be detected by both 3a- and caf1-based PCR reagents. In addition, a simple and efficient method for soil sample processing is presented here.Conclusions/SignificanceThe vacuum-dried reagents for real-time PCR maintain accuracy and reproducibility for at least 49 days at 37°C, indicating that they can be easily transported at room temperature for field application if the machine for performing real-time PCR is available. This dry reagent is of great significance for routine plague surveillance.

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Detection and Identification of Yersinia pestis by Polymerase Chain Reaction (PCR) Using Multiplex Primers

Cited by 38 publications

References 13 publications

Yersinia pestis and Bubonic Plague

Yersinia pestis and Bubonic Plague

Yersinia pestis

Ambient Stable Quantitative PCR Reagents for the Detection of Yersinia pestis

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