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Multicenter evaluation of PCR reagents for Francisella tularensis

Multicenter evaluation of PCR reagents for Francisella tularensis

Abstracts of the General Meeting of the American Society for Microbiology 103: C-173

The potential threat of biowarfare necessitates having a quick identification method for agents such as Francisella tularensis. Standard methods of identifying Francisella spp. require special media and a long incubation period for growth and identification. Consequently, the Armed Forces Institute of Pathology, Division of Microbiology, developed and optimized PCR reagents for three gene targets of Francisella tularensis. The genes targeted are TUL4, SOD, and FOPA. AFIP and 5 other laboratories nationwide tested wet and lyophilized reagents on the Ruggedized Advanced Pathogen Identification Device (RAPID)TM (Idaho Technology, Inc., Utah) for sensitivity and specificity of these assays. Reagents were evaluated by sending a panel of organisms consisting of multiple strains of Francisella tularensis, near neighbor organisms identified by 16S ribosomal relatedness, and positive and negative controls to the participating laboratories in the multicenter study. The results for the wet and lyophilized reagents were very similar. All reagents were able to identify 100% of samples containing Francisella tularensis DNA. The RAPID consistently detected 100 fg of DNA (approximately 50 gene copies). Swabs spiked with 104cfu/100ul of gamma irradiated Francisella tularensis SHU4 were also tested using the same targets. All laboratories participating in the study achieved 100% specificity with all three assays using the spiked swabs and the TUL4 lyophilized reagent achieved 100% sensitivity and specificity. Overall, the assays are reliable in identifying Francisella tularensis. The RAPID can identify Francisella tularensis in less than an hour after receiving the specimen allowing a quick turnaround time for patient diagnosis and treatment.

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Accession: 035349767

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