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Detection of Anthrax and Other Pathogens Using a Unique Liquid Array Technology
Author(s) -
Schweighardt Andrew J.,
Battaglia Amanda,
Wallace Margaret M.
Publication year - 2014
Publication title -
journal of forensic sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.715
H-Index - 96
eISSN - 1556-4029
pISSN - 0022-1198
DOI - 10.1111/1556-4029.12283
Subject(s) - ribosomal rna , microbiology and biotechnology , biology , hybridization probe , 23s ribosomal rna , bacteria , chemistry , chromatography , gene , rna , genetics , ribosome
A bead‐based liquid hybridization assay, L uminex ® 100™, was used to identify four pathogenic bacteria, B acillus anthracis , C lostridium botulinum , F rancisella tularensis subsp. tularensis , and Y ersinia pestis , and several close relatives. Hybridization between PCR ‐amplified target sequences and probe sequences (located within the 23S ribosomal RNA gene rrl and the genes related to the toxicity of each bacterium) was detected in single‐probe or multiple‐probe assays, depending on the organism. The lower limits of detection ( LLD s) for the probes ranged from 0.1 to 10 ng. Sensitivity was improved using lambda exonuclease to digest the noncomplementary target strand. All contributors in 33 binary, ternary, and quaternary mixtures in which all components were present in a 1:1 ratio were identified with an 80% success rate. Twenty‐eight binary mixtures in which the two components were combined in various ratios were further studied. All target sequences were detected, even when the minor component was overshadowed by a tenfold excess of the major component.