PDF(Pubmed)
Abstract：
Qualitative and quantitative PCR-based tests are widely used in both diagnostics and research to assess the prevalence of disease-causing pathogens in veterinary medicine. The efficacy of these tests, usually measured in terms of sensitivity and specificity, is critical in confirming or excluding a clinical diagnosis. We undertook a meta-analysis to assess the inherent value of published PCR diagnostic approaches used to confirm and quantify bacteria and viruses associated with bovine respiratory disease (BRD) in cattle. This review followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. A thorough search of nine electronic databases (Web of Science, EBSCOhost, Cambridge journals online, ProQuest, PubMed, Sage journals online, ScienceDirect, Wiley online library and MEDLINE) was undertaken to find studies that had reported on the use of PCR and/or qPCR for the detection and/or quantification of BRD associated organisms. All studies meeting the inclusion criteria for reporting quantitative PCR for identification of BRD associated microorganisms were included in the analysis. Studies were then assessed on the applications of the Minimum Information for Publication of Quantitative Real-Time PCR Experiment (MIQE) and PCR primer/probe sequences were extracted and tested for in silico specificity using a high level of stringency. Fourteen full-text articles were included in this study. Of these, 79% of the analysed articles did not report the application of the MIQE guidelines in their study. High stringency in silico testing of 144 previously published PCR primer/probe sequences found many to have questionable specificity. This review identified a high occurrence of primer/probe sequences with a variable in silico specificity such that this may have implications for the accuracy of reporting. Although this analysis was only applied to one specific disease state, identification of animals suspected to be suffering from bovine respiratory disease, there appears to be more broadly a need for veterinary diagnostic studies to adopt international best practice for reporting of quantitative PCR diagnostic data to be both accurate and comparable between studies and methodologies.
摘要：
基于定性和定量PCR的测试广泛用于诊断和研究，以评估兽医学中致病病原体的患病率。这些测试的功效，通常以灵敏度和特异性来衡量，对确认或排除临床诊断至关重要。我们进行了荟萃分析，以评估已发表的PCR诊断方法的内在价值，该方法用于确认和定量与牛呼吸道疾病（BRD）相关的细菌和病毒。本审查遵循了系统审查和荟萃分析（PRISMA）指南的首选报告项目。对九个电子数据库的彻底搜索(WebofScience，EBSCOhost,剑桥在线期刊，ProQuest,PubMed,Sage在线期刊，ScienceDirect,Wiley在线图书馆和MEDLINE）进行了研究，以发现有关使用PCR和/或qPCR检测和/或定量BRD相关生物的研究。所有符合报告定量PCR以鉴定BRD相关微生物的纳入标准的研究都包括在分析中。然后对定量实时PCR实验（MIQE）的最低信息的应用进行评估，并提取PCR引物/探针序列，并使用高水平的严格性测试计算机特异性。本研究共纳入14篇全文。其中,79%的分析文章没有报告MIQE指南在他们的研究中的应用。在144个先前公开的PCR引物/探针序列的计算机模拟测试中的高严格性发现许多具有可疑的特异性。这篇评论确定了具有可变的计算机特异性的引物/探针序列的高发生率，因此这可能对报告的准确性有影响。尽管此分析仅适用于一种特定的疾病状态，怀疑患有牛呼吸道疾病的动物的鉴定，兽医诊断研究似乎更广泛地需要采用国际最佳实践来报告定量PCR诊断数据,使其在研究和方法之间既准确又具有可比性.
