Detection of Borrelia burgdorferi infection in Ixodes dammini ticks with the polymerase chain reaction

Abstract

The polymerase chain reaction (PCR) was used to amplify DNA sequences of the etiologic agent of Lyme disease, Borrelia burgdorferi, and was applied to the detection of the spirochete in its tick vector. The target for PCR amplification was the OSP-A gene of strain B31; analysis of isolates from different geographical areas indicated that this gene could be used to identify most North American isolates. These methods were extended to the analysis of colony-derived and field-collected Ixodes dammini. OSP-A-specific sequences were identified in 15 of 15 colony-derived nymphal ticks that had fed previously on an infected animal; no such amplification products were detected in 8 control ticks. Segregated midgut tissues of field-collected adult and nymphal ticks from Nantucket Island, Mass., and the Crane Reserve, Ipswich, Mass., were examined by both direct fluorescent-antibody (DFA) staining and PCR. The DFA technique identified 16 infected ticks of 30 paired specimens; 15 of these specimens were positive by PCR. One specimen was positive by PCR that was DFA negative. Both live whole ticks and desiccated dead specimens were suitable for this analysis. Because only live ticks are suitable for DFA analysis, the use of PCR may extend the range of specimens that can be analyzed for the presence of the Lyme spirochete.