A multiplex polymerase chain reaction microarray assay to detect bioterror pathogens in blood.
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.
Tomioka
Keiko
K
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.
Peredelchuk
Michael
M
Zhu
Xiangyang
X
Arena
Roberto
R
Volokhov
Dmitri
D
Selvapandiyan
Angamuthu
A
Stabler
Katie
K
Mellquist-Riemenschneider
Jenny
J
Chizhikov
Vladimir
V
Kaplan
Gerardo
G
Nakhasi
Hira
H
Duncan
Robert
R
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