Abstract：
Microbial source tracking (MST) technology represents an innovative approach employed to trace fecal contamination in environmental water systems. The performance of primers may be affected by amplification techniques, target primer categories, and regional differences. To investigate the influence of these factors on primer recognition performance, a meta-analysis was conducted on the application of MST in water environments using three databases: Web of Science, Scopus, and PubMed (n = 2291). After data screening, 46 studies were included in the final analysis. The investigation encompassed Polymerase Chain Reaction (PCR)/quantitative PCR (qPCR) methodologies, dye-based (SYBR)/probe-based (TaqMan) techniques, and geographical differences of a human host-specific (HF183) primer and other 21 additional primers. The results indicated that the primers analyzed were capable of differentiating host specificity to a certain degree. Nonetheless, by comparing sensitivity and specificity outcomes, it was observed that virus-based primers exhibited superior specificity and recognition capacity, as well as a stronger correlation with human pathogenicity in water environments compared to bacteria-based primers. This finding highlights an important direction for future advancements. Moreover, within the same category, qPCR did not demonstrate significant benefits over conventional PCR amplification methods. In comparing dye-based and probe-based techniques, it was revealed that the probe-based method\'s advantage lay primarily in specificity, which may be associated with the increased propensity of dye-based methods to produce false positives. Furthermore, the heterogeneity of the HF183 primer was not detected in China, Canada, and Singapore respectively, indicating a low likelihood of regional differences. The variation among the 21 other primers may be attributable to regional differences, sample sources, detection techniques, or alternative factors. Finally, we identified that economic factors, climatic conditions, and geographical distribution significantly influence primer performance.
摘要：
微生物源追踪（MST）技术代表了一种用于追踪环境水系统中粪便污染的创新方法。引物的性能可能受到扩增技术的影响，目标引物类别，和地区差异。为了研究这些因素对引物识别性能的影响,使用三个数据库对MST在水环境中的应用进行了荟萃分析：WebofScience，Scopus,和PubMed(n=2291)。经过数据筛选,46项研究纳入最终分析。调查包括聚合酶链反应（PCR）/定量PCR（qPCR）方法，基于染料（SYBR）/基于探针（TaqMan）的技术，以及人类宿主特异性（HF183）引物和其他21种其他引物的地理差异。结果表明，所分析的引物能够在一定程度上区分宿主特异性。尽管如此，通过比较敏感性和特异性结果，观察到基于病毒的引物表现出优异的特异性和识别能力，与基于细菌的引物相比，在水环境中与人类致病性的相关性更强。这一发现突出了未来发展的重要方向。此外，在同一类别中，qPCR未显示优于常规PCR扩增方法的显著益处。在比较基于染料和基于探针的技术时，研究表明，基于探针的方法的优势主要在于特异性，这可能与基于染料的方法产生假阳性的倾向增加有关。此外，在中国未检测到HF183引物的异质性，加拿大,分别是新加坡,表明区域差异的可能性很低。21种其他引物之间的差异可能归因于区域差异，样本来源，检测技术，或替代因素。最后,我们确定了经济因素，气候条件,和地理分布显著影响引物性能。
