Department of Large Animal Clinical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, SK, Canada.
Zoonoses Public Health. 2010 Nov;57 Suppl 1:115-20. doi: 10.1111/j.1863-2378.2010.01365.x.
The study objective was to evaluate the accuracy of a real-time polymerase chain reaction (RT-PCR) and a culture protocol used to detect Salmonella in the faeces of grow-finish pigs using a Bayesian approach. The RT-PCR was invA-gene-based assay, while the culture protocol included pre-enrichment in buffered peptone water, selective enrichment in tetrathionate and Rappaport-Vassiliadis broths, and isolation on semi-solid (modified semi-solid RV) or solid (XLT4, Rambach) agar plates. Bayesian analysis was performed using a two-test, two-population model with dependence between culture and RT-PCR and compared to a second model with conditional independence between these two tests. Two hundred and ninety three individual faecal and 294 pooled pen samples from grow-finish pig collected from 10 farms were tested and results were divided into two groups according to herd size (five herds <250 sows, five herds with >400 sows). In the dependence model, RT-PCR sensitivity (Se) and specificity (Sp) were estimated to be 90% (95% probability interval 74, 97) and 99% (98, 99), respectively. Culture Se was 92% (75, 99), while culture Sp was considered 100% as all culture-positive samples were confirmed by serotyping. In the conditional independence model, RT-PCR Se and Sp, and culture Se, were 96% (93, 98), 99% (98, 100) and 97% (94, 100), respectively. The dependence model resulted in posterior estimates of Se that were lower and with broader probability intervals than the independence model, indicating that when RT-PCR and culture are evaluated relative to each other, the correlation between these tests is an important source of bias and should be adjusted for during analysis. The RT-PCR evaluated in this study performed almost comparably to culture; given the cost savings associated with using this test and more timely results, the RT-PCR may be a useful alternative to culture for screening large numbers of samples, particularly when Salmonella prevalence is low.
本研究旨在采用贝叶斯方法评估实时聚合酶链反应(RT-PCR)和一种用于检测生长育肥猪粪便中沙门氏菌的培养方案的准确性。RT-PCR 基于 invA 基因检测,而培养方案包括在缓冲蛋白胨水中预增菌、在四硫代硫酸钠和 Rappaport-Vassiliadis 肉汤中选择性增菌,以及在半固体(改良半固体 RV)或固体(XLT4、Rambach)琼脂平板上分离。采用具有依赖性的两测试、两群体贝叶斯分析模型,比较了这两种测试之间条件独立的第二个模型。对来自 10 个农场的 293 份个体粪便和 294 份猪栏混合样本进行了检测,根据猪群规模(5 个猪群<250 头母猪,5 个猪群>400 头母猪)将结果分为两组。在依赖模型中,RT-PCR 的敏感性(Se)和特异性(Sp)估计分别为 90%(95%概率区间为 74%,97%)和 99%(98%,99%)。培养的 Se 为 92%(75%,99%),而培养的 Sp 被认为是 100%,因为所有培养阳性样本均通过血清分型得到确认。在条件独立模型中,RT-PCR 的 Se 和 Sp,以及培养的 Se,分别为 96%(93%,98%)、99%(98%,100%)和 97%(94%,100%)。依赖模型的 Se 后验估计值低于独立性模型,概率区间也较宽,这表明在评估 RT-PCR 和培养时,这两种测试之间的相关性是一个重要的偏倚来源,在分析中应进行调整。本研究评估的 RT-PCR 与培养相当;鉴于使用该检测的成本节约以及更及时的结果,当沙门氏菌流行率较低时,RT-PCR 可能是一种用于筛选大量样本的有用替代方法,特别是在检测时。