Development of a predictive model for detection of Mycobacterium avium subsp. paratuberculosis in faeces by quantitative real time PCR.
摘要：
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.
收起
展开
DOI：
10.1016/j.vetmic.2010.10.009
被引量：
年份：
1970
ResearchGate
(全网免费下载)
钛学术
(全网免费下载)
通过 文献互助 平台发起求助，成功后即可免费获取论文全文。
求助方法1：
知识发现用户
每天可免费求助50篇
求助方法1：
关注微信公众号
每天可免费求助2篇
求助方法2：
完成求助需要支付5财富值
您目前有 1000 财富值
相似文献(220)
参考文献(0)
引证文献(24)
来源期刊
影响因子：暂无数据
JCR分区： 暂无
中科院分区：暂无
