Abstract
Epizootic bovine abortion (EBA), also commonly known as "foothill abortion," is a late-term abortion primarily in beef cattle with significant economic impacts in California, Nevada, and Oregon. The causative agent is a novel deltaproteobacterium (aoEBA) closely related to the order Myxococcales and vectored by the soft-shelled tick Ornithodoros coriaceus. Historically, diagnosis has relied upon the pathologic examination of the fetus and the presence of elevated fetal serum immunoglobulins. Identification of the etiologic agent, a unique deltaproteobacterium, permitted the development of a quantitative duplex real-time polymerase chain reaction (qPCR) using a unique 90-bp sequence of aoEBA 16S ribosomal RNA gene in conjunction with an 88-bp sequence of the bovine β-actin gene. Reaction efficiencies were 100.9% for the 16S aoEBA gene and 93.1% for the bovine β-actin gene. Application of the duplex TaqMan to a set of aoEBA-infected fetal bovine necropsy tissues demonstrated the assay to be robust in quantitatively identifying the aoEBA bacteria and establishing host-tissue pathogen load. Consistent with previously reported immunohistochemical data, organized lymphoid tissue generally carried the heaviest bacterial load as compared to non-lymphoid tissue. The newly developed duplex TaqMan assay will facilitate diagnosis in difficult cases and provide an invaluable tool for delineating the pathogenesis of EBA.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1153-1159 |
| Number of pages | 7 |
| Journal | Journal of Veterinary Diagnostic Investigation |
| Volume | 23 |
| Issue number | 6 |
| DOIs | |
| State | Published - Nov 2011 |
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Keywords
- Deltaproteobacteria
- epizootic bovine abortion
- polymerase chain reaction
ASJC Scopus subject areas
- veterinary(all)
Cite this
Quantitative duplex taqman real-time polymerase chain reaction for the assessment of the etiologic agent of epizootic bovine abortion. / Brooks, Roxann S.; Blanchard, Myra; Anderson, Mark L; Hall, Mark R.; Stott, Jeffrey L.
In: Journal of Veterinary Diagnostic Investigation, Vol. 23, No. 6, 11.2011, p. 1153-1159.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Quantitative duplex taqman real-time polymerase chain reaction for the assessment of the etiologic agent of epizootic bovine abortion
AU - Brooks, Roxann S.
AU - Blanchard, Myra
AU - Anderson, Mark L
AU - Hall, Mark R.
AU - Stott, Jeffrey L
PY - 2011/11
Y1 - 2011/11
N2 - Epizootic bovine abortion (EBA), also commonly known as "foothill abortion," is a late-term abortion primarily in beef cattle with significant economic impacts in California, Nevada, and Oregon. The causative agent is a novel deltaproteobacterium (aoEBA) closely related to the order Myxococcales and vectored by the soft-shelled tick Ornithodoros coriaceus. Historically, diagnosis has relied upon the pathologic examination of the fetus and the presence of elevated fetal serum immunoglobulins. Identification of the etiologic agent, a unique deltaproteobacterium, permitted the development of a quantitative duplex real-time polymerase chain reaction (qPCR) using a unique 90-bp sequence of aoEBA 16S ribosomal RNA gene in conjunction with an 88-bp sequence of the bovine β-actin gene. Reaction efficiencies were 100.9% for the 16S aoEBA gene and 93.1% for the bovine β-actin gene. Application of the duplex TaqMan to a set of aoEBA-infected fetal bovine necropsy tissues demonstrated the assay to be robust in quantitatively identifying the aoEBA bacteria and establishing host-tissue pathogen load. Consistent with previously reported immunohistochemical data, organized lymphoid tissue generally carried the heaviest bacterial load as compared to non-lymphoid tissue. The newly developed duplex TaqMan assay will facilitate diagnosis in difficult cases and provide an invaluable tool for delineating the pathogenesis of EBA.
AB - Epizootic bovine abortion (EBA), also commonly known as "foothill abortion," is a late-term abortion primarily in beef cattle with significant economic impacts in California, Nevada, and Oregon. The causative agent is a novel deltaproteobacterium (aoEBA) closely related to the order Myxococcales and vectored by the soft-shelled tick Ornithodoros coriaceus. Historically, diagnosis has relied upon the pathologic examination of the fetus and the presence of elevated fetal serum immunoglobulins. Identification of the etiologic agent, a unique deltaproteobacterium, permitted the development of a quantitative duplex real-time polymerase chain reaction (qPCR) using a unique 90-bp sequence of aoEBA 16S ribosomal RNA gene in conjunction with an 88-bp sequence of the bovine β-actin gene. Reaction efficiencies were 100.9% for the 16S aoEBA gene and 93.1% for the bovine β-actin gene. Application of the duplex TaqMan to a set of aoEBA-infected fetal bovine necropsy tissues demonstrated the assay to be robust in quantitatively identifying the aoEBA bacteria and establishing host-tissue pathogen load. Consistent with previously reported immunohistochemical data, organized lymphoid tissue generally carried the heaviest bacterial load as compared to non-lymphoid tissue. The newly developed duplex TaqMan assay will facilitate diagnosis in difficult cases and provide an invaluable tool for delineating the pathogenesis of EBA.
KW - Deltaproteobacteria
KW - epizootic bovine abortion
KW - polymerase chain reaction
UR - http://www.scopus.com/inward/record.url?scp=84857090231&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84857090231&partnerID=8YFLogxK
U2 - 10.1177/1040638711425573
DO - 10.1177/1040638711425573
M3 - Article
C2 - 22362796
AN - SCOPUS:84857090231
VL - 23
SP - 1153
EP - 1159
JO - Journal of Veterinary Diagnostic Investigation
JF - Journal of Veterinary Diagnostic Investigation
SN - 1040-6387
IS - 6
ER -