Detection of Bartonella bacilliformis by Real-Time PCR in Naturally Infected Sand Flies
UNIFORMED SERVICES UNIV OF THE HEALTH SCIENCES BETHESDA MD
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Evidence incriminating phlebotomine sand flies as vectors of Bartonella bacilliformis, which causes bartonellosis, has not been proven beyond doubt. Our research was designed to strengthen this hypothesis. In addition, we developed a Real- Time PCR method capable of detecting as little as 100fg of Bartonella DNA in sand flies. Twelve primers were designed using TaqMan ProbePrimer Data and tested. Lutzomyia verrucarum sand flies were collected from an endemic focus of bartonellosis in and around Caraz, Ancash, Peru. Unfed female sand flies were pooled in groups of 5 for DNA extraction. Of 472 pools assayed, 13 tested positive 2.75. The sensitivity of the test was 100 and specificity was 91.11 based on assays of known samples. We conclude that this method has the necessary sensitivity and specificity to detect B. bacilliformis infection in wild-caught sand flies enabling us to determine the true infection sand fly rates in an endemic area of Peru.