JMD
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Purchase Article
Right arrow View Shopping Cart
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Selvapandiyan, A.
Right arrow Articles by Nakhasi, H. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Selvapandiyan, A.
Right arrow Articles by Nakhasi, H. L.
JMD 2005, Vol. 7, No. 2
Copyright © 2005 American Society for Investigative Pathology & Association for Molecular Pathology

A Novel Semiquantitative Fluorescence-Based Multiplex Polymerase Chain Reaction Assay for Rapid Simultaneous Detection of Bacterial and Parasitic Pathogens from Blood

Angamuthu Selvapandiyan*, Katie Stabler{dagger}, Nasim A. Ansari{ddagger}, Stephen Kerby*, Jenny Riemenschneider{dagger}, Poonam Salotra{ddagger}, Robert Duncan* and Hira L. Nakhasi*

From the Divisions of Emerging and Transfusion Transmitted Disease * and Hematology, {dagger} Office of Blood Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland; and the Institute of Pathology (Indian Council of Medical Research), {ddagger} Safdarjung Hospital, New Delhi, India

A multiplex polymerase chain reaction assay was developed for the rapid simultaneous detection of category A select bacterial agents (Bacillus anthracis and Yersinia pestis) and parasitic pathogens (Leishmania species) in blood using the Cepheid Smart Cycler platform. B. anthracis (Sterne) and Yersinia. pseudotuberculosis were used in the assay for optimization for B. anthracis and Y. pestis, respectively. The specificity of the target amplicons [protective antigen gene of B. anthracis and rRNA genes of other pathogens or human (internal control)] was evaluated by staining the amplicons with SYBR Green I and determining their individual melting temperatures (Tm). As a novel approach for pathogen semiquantitation, the Tm peak height of the amplicon was correlated with a known standard curve of pathogen-spiked samples. This assay was able to detect DNA in blood spiked with less than 50 target cells/ml for all of the pathogens. The sensitivity of this assay in blood was 100% for the detection of Leishmania donovani from leishmaniasis patients and B. anthracis (Sterne) from symptomatic mice. The time necessary for performing this assay including sample preparation was less than 1.5 hours, making this a potentially useful method for rapidly diagnosing and monitoring the efficacy of drugs or vaccines in infected individuals.




This article has been cited by other articles:


Home page
 Am J Trop Med HygHome page
E. S. Gabitzsch, R. Vera-Tudela, R. J. Eisen, S. W. Bearden, K. L. Gage, and N. S. Zeidner
Development of a Real-time Quantitative PCR Assay to Enumerate Yersinia pestis in Fleas
Am J Trop Med Hyg, July 1, 2008; 79(1): 99 - 101.
[Abstract] [Full Text] [PDF]

Home page
 J. Mol. Diagn.Home page
K. Tomioka, M. Peredelchuk, X. Zhu, R. Arena, D. Volokhov, A. Selvapandiyan, K. Stabler, J. Mellquist-Riemenschneider, V. Chizhikov, G. Kaplan, et al.
A Multiplex Polymerase Chain Reaction Microarray Assay to Detect Bioterror Pathogens in Blood
J. Mol. Diagn., October 1, 2005; 7(4): 486 - 494.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2005 by the American Society for Investigative Pathology and the Association for Molecular Pathology.