Detection of hemoplasma infection of goats by use of a quantitative polymerase chain reaction assay and risk factor analysis for infection
| Autor: | G. Kenitra Hammac, Naíla C. do Nascimento, Kathy A. Johnson, Hsin-Yi Weng, Joanne B. Messick, Amy E. Bauer |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Indiana 040301 veterinary sciences Biology Real-Time Polymerase Chain Reaction Sensitivity and Specificity 0403 veterinary science 03 medical and health sciences Mycoplasma Plasmid Animals Mycoplasma Infections Risk factor Ovis DNA Primers Detection limit Goat Diseases General Veterinary Adult female Goats 04 agricultural and veterinary sciences General Medicine biology.organism_classification DNA extraction Virology Blood film 030104 developmental biology Real-time polymerase chain reaction Female Factor Analysis Statistical |
| Zdroj: | American Journal of Veterinary Research. 77:882-889 |
| ISSN: | 0002-9645 |
| DOI: | 10.2460/ajvr.77.8.882 |
| Popis: | OBJECTIVE To develop and validate a real-time quantitative PCR (qPCR) assay for the detection and quantification of Mycoplasma ovis in goats and investigate the prevalence and risk factors for hemoplasma infection of goats located in Indiana. ANIMALS 362 adult female goats on 61 farms. PROCEDURES Primers were designed for amplification of a fragment of the dnaK gene of M ovis by use of a qPCR assay. Blood samples were collected into EDTA-containing tubes for use in total DNA extraction, blood film evaluation, and determination of PCV. Limit of detection, intra-assay variability, interassay variability, and specificity of the assay were determined. RESULTS Reaction efficiency of the qPCR assay was 94.45% (R2, 0.99; slope, −3.4623), and the assay consistently detected as few as 10 copies of plasmid/reaction. Prevalence of infection in goats on the basis of results for the qPCR assay was 18.0% (95% confidence interval, 14% to 22%), with infected goats ranging from 1 to 14 years old, whereby 61% (95% confidence interval, 47% to 73%) of the farms had at least 1 infected goat. Bacterial load in goats infected with M ovis ranged from 1.05 × 103 target copies/mL of blood to 1.85 × 105 target copies/mL of blood; however, no bacteria were observed on blood films. Production use of a goat was the only risk factor significantly associated with hemoplasma infection. CONCLUSIONS AND CLINICAL RELEVANCE The qPCR assay was more sensitive for detecting hemoplasma infection than was evaluation of a blood film, and production use of a goat was a risk factor for infection. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|