Abstract
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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1509-1515 |
| Number of pages | 7 |
| Journal | American Journal of Veterinary Research |
| Volume | 55 |
| Issue number | 11 |
| State | Published - Nov 1994 |
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Polymerase chain reaction for detection of Borrelia coriaceae, putative agent of epizootic bovine abortion. / Zingg, B. C.; Lefebvre, Rance B.
In: American Journal of Veterinary Research, Vol. 55, No. 11, 11.1994, p. 1509-1515.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Polymerase chain reaction for detection of Borrelia coriaceae, putative agent of epizootic bovine abortion.
AU - Zingg, B. C.
AU - Lefebvre, Rance B
PY - 1994/11
Y1 - 1994/11
N2 - 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.
AB - 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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UR - http://www.scopus.com/inward/citedby.url?scp=0028538550&partnerID=8YFLogxK
M3 - Article
VL - 55
SP - 1509
EP - 1515
JO - American Journal of Veterinary Research
JF - American Journal of Veterinary Research
SN - 0002-9645
IS - 11
ER -