Polymerase chain reaction for detection of Borrelia coriaceae, putative agent of epizootic bovine abortion

Barbara C. Zingg From the Department of Veterinary Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616.

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Rance B. LeFebvre From the Department of Veterinary Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616.

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Summary

The nucleotide sequence of a chromosomally encoded antigen-expressing gene of Borrelia coriaceae was determined and used as a target for the polymerase chain reaction (PCR). Two primer sets were designed specifying the amplification of 269- and 701-bp dna fragments. Primer set I, producing the short amplicon, was tenfold more sensitive than primer set II. As little as 10 fg of purified B coriaceae dna could consistently be detected. The pcr assays, containing controlled numbers of whole spirochetes, allowed detectable amplification of 2 to 10 organisms. An internal, nonradioactively labeled gene-specific probe verified specificity of the pcr amplicons. Neither primer set cross-reacted with other related spirochetes. This pcr assay was adapted and found suitable for identification of B coriaceae in biological samples, such as blood and thymus. Evidence for presence of B coriaceae in biological samples was not found in tissue samples obtained from experimentally infected cows and their fetuses. These data failed to establish a definite association between B coriaceae and epizootic bovine abortion.

Summary

The nucleotide sequence of a chromosomally encoded antigen-expressing gene of Borrelia coriaceae was determined and used as a target for the polymerase chain reaction (PCR). Two primer sets were designed specifying the amplification of 269- and 701-bp dna fragments. Primer set I, producing the short amplicon, was tenfold more sensitive than primer set II. As little as 10 fg of purified B coriaceae dna could consistently be detected. The pcr assays, containing controlled numbers of whole spirochetes, allowed detectable amplification of 2 to 10 organisms. An internal, nonradioactively labeled gene-specific probe verified specificity of the pcr amplicons. Neither primer set cross-reacted with other related spirochetes. This pcr assay was adapted and found suitable for identification of B coriaceae in biological samples, such as blood and thymus. Evidence for presence of B coriaceae in biological samples was not found in tissue samples obtained from experimentally infected cows and their fetuses. These data failed to establish a definite association between B coriaceae and epizootic bovine abortion.

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