School of Civil Engineering, Sun Yat-Sen University, Zhuhai, 519082, China; Guangdong Key Laboratory of Integrated Agro-environmental Pollution Control and Management, Institute of Eco-environmental and Soil Sciences, Guangdong Academy of Sciences, Guangzhou, 510070, China.
School of Civil Engineering, Sun Yat-Sen University, Zhuhai, 519082, China.
J Environ Manage. 2023 Nov 1;345:118800. doi: 10.1016/j.jenvman.2023.118800. Epub 2023 Aug 15.
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.
微生物源追踪 (MST) 技术代表了一种创新性的方法，用于追踪环境水系中的粪便污染。引物的性能可能会受到扩增技术、目标引物类别和区域差异的影响。为了研究这些因素对引物识别性能的影响，我们对使用三个数据库（Web of Science、Scopus 和 PubMed）的 MST 在水环境中的应用进行了荟萃分析（n=2291）。经过数据筛选，最终有 46 项研究纳入了分析。调查涵盖了聚合酶链反应 (PCR)/定量 PCR (qPCR) 方法、染料基 (SYBR)/探针基 (TaqMan) 技术，以及人类宿主特异性 (HF183) 引物和其他 21 个额外引物的地理差异。结果表明，所分析的引物在一定程度上能够区分宿主特异性。然而，通过比较敏感性和特异性结果，观察到基于病毒的引物在环境水中具有更好的特异性和识别能力，并且与人类致病性的相关性更强，而基于细菌的引物则不然。这一发现为未来的发展指明了一个重要方向。此外，在相同类别中，qPCR 并未显示出比传统 PCR 扩增方法有显著优势。在比较染料基和探针基技术时，发现探针基方法的优势主要在于特异性，这可能与染料基方法产生假阳性的倾向增加有关。此外，HF183 引物在中国、加拿大和新加坡分别没有检测到异质性，表明区域差异的可能性较低。其他 21 个引物的变异可能归因于区域差异、样本来源、检测技术或其他因素。最后，我们发现经济因素、气候条件和地理分布显著影响引物性能。
