Barnewall Rebecca J, Marsh Ian B, Quinn Jane C
School of Agricultural, Environmental and Veterinary Science, Charles Sturt University, Wagga Wagga, NSW, Australia.
Gulbali Institute, Wagga Wagga, NSW, Australia.
Front Mol Biosci. 2022 Jul 18;9:902401. doi: 10.3389/fmolb.2022.902401. eCollection 2022.
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 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 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 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.
基于定性和定量PCR的检测方法在兽医学的诊断和研究中被广泛用于评估致病病原体的流行情况。这些检测方法的有效性通常以灵敏度和特异性来衡量,对于确认或排除临床诊断至关重要。我们进行了一项荟萃分析,以评估已发表的用于确认和量化与牛呼吸道疾病(BRD)相关的细菌和病毒的PCR诊断方法的内在价值。本综述遵循系统评价和荟萃分析的首选报告项目(PRISMA)指南。我们对九个电子数据库(科学网、EBSCOhost、剑桥在线期刊、ProQuest、PubMed、Sage在线期刊、ScienceDirect、Wiley在线图书馆和MEDLINE)进行了全面搜索,以查找报告使用PCR和/或qPCR检测和/或量化与BRD相关生物的研究。所有符合报告定量PCR用于鉴定BRD相关微生物的纳入标准的研究都纳入了分析。然后根据定量实时PCR实验发表的最低信息(MIQE)的应用对研究进行评估,并提取PCR引物/探针序列,使用严格的标准测试其特异性。本研究纳入了14篇全文文章。其中,79%的分析文章在其研究中未报告MIQE指南的应用。对144个先前发表的PCR引物/探针序列进行的严格测试发现,许多序列的特异性存在问题。本综述发现引物/探针序列特异性差异很大的情况很常见,这可能会影响报告的准确性。尽管该分析仅适用于一种特定疾病状态,即疑似患有牛呼吸道疾病的动物的鉴定,但似乎更广泛地需要兽医诊断研究采用国际最佳实践来报告定量PCR诊断数据,以便研究和方法之间的数据准确且具有可比性。