Polymerase chain reaction for detection of Borrelia coriaceae, putative agent of epizootic bovine abortion.
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.
Zingg
B C
BC
Department of Veterinary Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis 95616.
LeFebvre
R B
RB
D000034:Abortion, Veterinary / Q000097:blood / Q000175:diagnosis / Q000382:microbiology*; D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000235:genetics*; D001483:Base Sequence; D001769:Blood / Q000382:microbiology; D001898:Borrelia / Q000235:genetics / Q000302:isolation & purification*; D001899:Borrelia Infections / Q000097:blood / Q000175:diagnosis / Q000662:veterinary*; D002876:Chromosomes, Bacterial; D003429:Cross Reactions; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D005260:Female; D005312:Fetal Blood / Q000382:microbiology; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D011247:Pregnancy; D012680:Sensitivity and Specificity; D013950:Thymus Gland / Q000382:microbiology*
D000942:Antigens, Bacterial; D017931:DNA Primers; D004269:DNA, Bacterial