title: Reverse line blot macroarray for simultaneous detection and characterization of four biological warfare agents.
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(2) cfu/ml for B. anthracis, 8 × 10(2) cfu/ml for Yersinia sp., 1.4 × 10(2) cfu/ml for B. melitensis and 4 × 10(2) cfu/ml for B. pseudomallei.
authors.affiliation: Division of Microbiology, Defence Research and Development Establishment, Jhansi Road, Gwalior, 474002 India.
authors.lastname: Vanlalhmuaka
authors.affiliation: Division of Microbiology, Defence Research and Development Establishment, Jhansi Road, Gwalior, 474002 India.
authors.forename: Kulanthaivel
authors.initials: K
authors.lastname: Thavachelvam
authors.affiliation: Division of Microbiology, Defence Research and Development Establishment, Jhansi Road, Gwalior, 474002 India.
authors.forename: Urmil
authors.initials: U
authors.lastname: Tuteja
authors.affiliation: Division of Microbiology, Defence Research and Development Establishment, Jhansi Road, Gwalior, 474002 India.
authors.forename: Kumari
authors.initials: K
authors.lastname: Sarika
authors.affiliation: Division of Microbiology, Defence Research and Development Establishment, Jhansi Road, Gwalior, 474002 India.
authors.forename: Suryanarayana
authors.initials: S
authors.lastname: Nagendra
authors.affiliation: Division of Microbiology, Defence Research and Development Establishment, Jhansi Road, Gwalior, 474002 India.
authors.forename: Subodh
authors.initials: S
authors.lastname: Kumar
keywords: Bacillus anthracis; Biowarfare; Brucella; Burkholderia; Reverse line blot; Yersinia pestis