Quantitative duplex TaqMan real-time polymerase chain reaction for the assessment of the etiologic agent of epizootic bovine abortion.
J Vet Diagn Invest
; 23(6): 1153-9, 2011 Nov.
Article
in En
| MEDLINE
| ID: mdl-22362796
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 ß-actin gene. Reaction efficiencies were 100.9% for the 16S aoEBA gene and 93.1% for the bovine ß-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.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Deltaproteobacteria
/
Abortion, Veterinary
/
Real-Time Polymerase Chain Reaction
Type of study:
Etiology_studies
/
Prognostic_studies
Limits:
Animals
Language:
En
Journal:
J Vet Diagn Invest
Journal subject:
MEDICINA VETERINARIA
Year:
2011
Document type:
Article
Affiliation country:
United States