Development of a predictive model for detection of Mycobacterium avium subsp. paratuberculosis in faeces by quantitative real time PCR
Language English Country Netherlands Media print-electronic
Document type Comparative Study, Journal Article, Research Support, Non-U.S. Gov't
PubMed
21075565
DOI
10.1016/j.vetmic.2010.10.009
PII: S0378-1135(10)00496-7
Knihovny.cz E-resources
- MeSH
- DNA, Bacterial analysis isolation & purification MeSH
- Feces microbiology MeSH
- Limit of Detection MeSH
- Logistic Models * MeSH
- Mycobacterium avium subsp. paratuberculosis genetics isolation & purification MeSH
- Cattle Diseases diagnosis microbiology MeSH
- Paratuberculosis diagnosis microbiology MeSH
- Polymerase Chain Reaction methods veterinary MeSH
- Sensitivity and Specificity MeSH
- Cattle MeSH
- Animals MeSH
- Check Tag
- Cattle MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Comparative Study MeSH
- Names of Substances
- DNA, Bacterial MeSH
This study focused on the development of a reliable and cost-efficient DNA isolation procedure for the detection of Mycobacterium avium subsp. paratuberculosis (MAP) in faeces by previously developed IS900 and F57 quantitative real time PCR (qPCR) and their comparison with culture. The recovery of MAP DNA from the spiking experiments ranged from 29.1 to 102.4% of the input amount of MAP with median 37.9%. The limit of detection was determined to be 1.03 × 10(4) for F57 qPCR and 6.87 × 10(2)MAP cells per gram of faeces for IS900 qPCR, respectively. The developed technique for DNA isolation was coupled with IS900 qPCR and compared to traditional MAP culture using a cohort of 1906 faecal samples examined from 12 dairy cattle farms in our laboratory. From those 1906 original faecal samples, 875 were positive by IS900 qPCR and 169 by culture. None of the culture positive samples was negative by IS900 qPCR. This data facilitated development of a predictive model capable of estimating the probability of being culture positive by estimating the absolute number of MAP per gram of faeces as determined IS900 qPCR without performing the culture.
References provided by Crossref.org
Recovery of Mycobacteria from Heavily Contaminated Environmental Matrices
Utilization of Digital PCR in Quantity Verification of Plasmid Standards Used in Quantitative PCR
A Basic Guide to Real Time PCR in Microbial Diagnostics: Definitions, Parameters, and Everything
Influence of Stress Connected with Moving to a New Farm on Potentially MAP-Infected Mouflons
