title: A multiplex polymerase chain reaction microarray assay to detect bioterror pathogens in blood.
abstract: Heightened concern about the dangers of bioterrorism requires that measures be developed to ensure the safety of the blood supply. Multiplex detection of such agents using a blood-screening DNA microarray is a sensitive and specific method to screen simultaneously for a number of suspected agents. We have developed and optimized a multiplex polymerase chain reaction microarray assay to screen blood for three potential bioterror bacterial pathogens and a human ribosomal RNA gene internal control. The analytical sensitivity of the assay was demonstrated to be 50 colony-forming units/ml for Bacillus anthracis, Francisella tularensis, and Yersinia pseudotuberculosis (surrogate for Yersinia pestis). The absence of any false-positives demonstrated high analytical specificity. Screening B. anthracis-infected mouse blood samples and uninfected controls demonstrated effectiveness and specificity in a preclinical application. This study represents proof of the concept of microarray technology to screen simultaneously for multiple bioterror pathogens in blood samples.
authors.affiliation: Division of Emerging and Transfusion Transmitted Diseases, Office of Blood Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD, USA.
authors.forename: Keiko
authors.initials: K
authors.lastname: Tomioka
authors.forename: Michael
authors.initials: M
authors.lastname: Peredelchuk
authors.forename: Xiangyang
authors.initials: X
authors.lastname: Zhu
authors.forename: Roberto
authors.initials: R
authors.lastname: Arena
authors.forename: Dmitri
authors.initials: D
authors.lastname: Volokhov
authors.forename: Angamuthu
authors.initials: A
authors.lastname: Selvapandiyan
authors.forename: Katie
authors.initials: K
authors.lastname: Stabler
authors.forename: Jenny
authors.initials: J
authors.lastname: Mellquist-Riemenschneider
authors.forename: Vladimir
authors.initials: V
authors.lastname: Chizhikov
authors.forename: Gerardo
authors.initials: G
authors.lastname: Kaplan
authors.forename: Hira
authors.initials: H
authors.lastname: Nakhasi
authors.forename: Robert
authors.initials: R
authors.lastname: Duncan
mesh_terms: D000818:Animals; D000881:Anthrax / Q000097:blood* / Q000175:diagnosis* / Q000382:microbiology; D001408:Bacillus anthracis / Q000235:genetics* / Q000302:isolation & purification*; D023482:Bioterrorism; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction