Simultaneous Detection of CDC Category A DNA and RNA Bioterrorism Agents by Use of Multiplex PCR RT-PCR Enzyme Hybridization Assays 英文参考文献.docVIP

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Simultaneous Detection of CDC Category A DNA and RNA Bioterrorism Agents by Use of Multiplex PCR RT-PCR Enzyme Hybridization Assays 英文参考文献.doc

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SimultaneousDetectionofCDCCategoryADNAandRNABioterrorismAgentsbyUseofMultiplexPCR

Viruses 2009, 1, 441-459; doi:10.3390/v1030441 OPEN ACCESS viruses ISSN 1999-4915 /journal/viruses Article Simultaneous Detection of CDC Category A DNA and RNA Bioterrorism Agents by Use of Multiplex PCR RT-PCR Enzyme Hybridization Assays Jie He 1, Andrea J. Kraft 1, Jiang Fan 1, Meredith Van Dyke 1, Lihua Wang 1, Michael E. Bose 1, Marilyn Khanna 2, Jacob A. Metallo 1 and Kelly J. Henrickson 1,3,4,* 1 Department of Pediatric, Medical College of Wisconsin, Milwaukee, WI 53226, USA; E-Mails: jhe@ (J.H.); akraft@ (A.J.K.); jfan@ (J.F.); mvandyke@ (M.V.D.); lwang@ (L.W.); mbose@ (M.E.B.); jmetallo@ (J.A.M.) 2 National Institutes of Health, Bethesda, MD 20892-7616, USA; E-Mail: khannama@ (M.K.) 3 Children’s Research Institute, Children’s Hospital of Wisconsin, P.O. Box 1997, Milwaukee, WI 53201-1997, USA 4 Children’s Hospital of Wisconsin, P.O. Box 1997, Milwaukee, WI 53201-1997, USA * Author to whom correspondence should be addressed; E-Mail: khenrick@; Tel.: +1-414-337-7073; Fax: +1-414-337-7093. Received: 13 July 2009; in revised form: 22 September 2009 / Accepted: 20 October 2009 / Published: 20 October 2009 Abstract: Assays to simultaneously detect multiple potential agents of bioterrorism are limited. Two multiplex PCR and RT-PCR enzyme hybridization assays (mPCR-EHA, mRT-PCR-EHA) were developed to simultaneously detect many of the CDC category “A” bioterrorism agents. The “Bio T” DNA assay was developed to detect: Variola major (VM), Bacillus anthracis (BA), Yersinia pestis (YP), Francisella tularensis (FT) and Varicella zoster virus (VZV). The “Bio T” RNA assay (mRT-PCR-EHA) was developed to detect: Ebola virus (Ebola), Lassa fever virus (Lassa), Rift Valley fever (RVF), Hantavirus Sin Nombre species (HSN) and dengue virus (serotypes 1-4). Sensitivity and specificity of the 2 assays were tested by using genomic DNA, recombinant plasmid positive controls, RNA transcripts controls, surrogate (spiked) clinic