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JMD 2005, Vol. 7, No. 4
Copyright © 2005 American Society for Investigative Pathology & Association for Molecular Pathology

A Multiplex Polymerase Chain Reaction Microarray Assay to Detect Bioterror Pathogens in Blood

Keiko Tomioka*, Michael Peredelchuk*, Xiangyang Zhu*, Roberto Arena*, Dmitri Volokhov{dagger}, Angamuthu Selvapandiyan*, Katie Stabler{ddagger}, Jenny Mellquist-Riemenschneider{ddagger}, Vladimir Chizhikov{dagger}, Gerardo Kaplan*, Hira Nakhasi* and Robert Duncan*

From the Divisions of Emerging and Transfusion Transmitted Diseases * and Hematology, {ddagger} Office of Blood Research and Review, and the Division of Viral Products, {dagger} Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland

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.




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[Abstract] [Full Text] [PDF]


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