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- Title
Reverse Line Blot Macroarray for Simultaneous Detection and Characterization of Four Biological Warfare Agents.
- Authors
Vanlalhmuaka; Thavachelvam, Kulanthaivel; Tuteja, Urmil; Sarika, Kumari; Nagendra, Suryanarayana; Kumar, Subodh
- Abstract
The need for a rapid detection and characterization of biowarfare (BW) agents cannot be over emphasized. With diverse array of potential BW pathogen available presently, rapid identification of the pathogen is crucial, so that specific therapy and control measures can be initiated. We have developed a multiplex polymerase chain reaction based reverse line blot macroarray to simultaneously detect four pathogens of BW importance viz. Bacillus anthracis, Yersinia pestis, Brucella melitensis and Burkholderia pseudomallei. The multiplex PCR utilizes 14 pairs of primers targeting 18 specific markers. These markers include genes which are genus specific, species-specific chromosomal sequences and virulence markers of plasmid origin. The assay was evaluated on various human, environment and animal isolates. The assay w successful in simultaneous detection and characterization of isolates of the four pathogens on as a single platform with sensitivity ranging from 0.3 pg to 0.3 ng of genomic DNA. The assay was able to detect 5 × 10 cfu/ml for B. anthracis, 8 × 10 cfu/ml for Yersinia sp., 1.4 × 10 cfu/ml for B. melitensis and 4 × 10 cfu/ml for B. pseudomallei.
- Subjects
IDENTIFICATION of pathogenic microorganisms; BACILLUS anthracis; YERSINIA pestis; BRUCELLA; BURKHOLDERIA; BIOLOGICAL warfare research; BIOLOGICAL weapons research
- Publication
Indian Journal of Microbiology, 2013, Vol 53, Issue 1, p41
- ISSN
0046-8991
- Publication type
Article
- DOI
10.1007/s12088-012-0330-7