Validation of host-specific Bacteroidales quantitative PCR assays and their application to microbial source tracking of drinking water sources in the Kathmandu Valley, Nepal
Validation of host-specific Bacteroidales quantitative PCR assays and their application to microbial source tracking of drinking water sources in the Kathmandu Valley, Nepal
复制标题
DOI：
10.1111/jam.13884
复制
发表时间：
2018-08-01
影响因子：
4
通讯作者：
Haramoto, E.
中科院分区：
文献类型：
--
作者：
Malla, B.;Shrestha, R. Ghaju;Haramoto, E.
目的验证宿主特异性拟杆菌目检测方法在尼泊尔加德满都山谷饮用水源中的粪源污染。方法和结果共收集了54份来自人类污水、反刍动物、猪、狗、鸡和鸭的复合粪源样品，采用定量聚合酶链反应（PCR）方法，（巴丘姆、HF 183 SYBR、gyrB和HF 183 TaqMan）、反刍动物特异性（BacCow和BacR）、猪特异性（Pig2Bac和PF 163）和狗特异性测定（BacCan SYBR）。巴丘姆、BacR和Pig2Bac测定分别被判定为最佳的人特异性、反刍动物特异性和猪特异性测定。BacCan SYBR检测试剂盒与其他菌种高度交叉反应，导致性能较差。此外，这些验证的方法被应用于74个饮用水样品的微生物源跟踪（MST）。其中，20%、12%和4%的样本分别被判定为受人类、反刍动物和猪粪便污染。人类和反刍动物粪便标志物的检出率相对较高，在建成区和农业arease.ConclusionBacterium，BacR和Pig2Bac检测被发现适合MST和两个，人类和动物粪便污染的饮用水源是常见的value.Significance和Impact的studyMST可能是一个有效的工具，为准备粪便污染的策略，因为这些是网站特定的。
AimsTo validate host-specific Bacteroidales assays to identify faecal-source contamination of drinking water sources in the Kathmandu Valley, Nepal.Methods and ResultsA total of 54 composite faecal-source samples were collected from human sewage, ruminants, pigs, dogs, chickens and ducks, which were analysed by quantitative polymerase chain reaction using human-specific (BacHum, HF183 SYBR, gyrB and HF183 TaqMan), ruminant-specific (BacCow and BacR), pig-specific (Pig2Bac and PF163) and dog-specific assays (BacCan SYBR). The BacHum, BacR and Pig2Bac assays were judged the best performing human-specific, ruminant-specific and pig-specific assays respectively. The BacCan SYBR assay highly cross-reacted with other species, resulting in poor performance. Furthermore, these validated assays were applied to microbial source tracking (MST) of 74 drinking water samples. Out of these, 20, 12 and 4% samples were judged contaminated by human, ruminant and pig faeces respectively. Detection ratios of human and ruminant faecal markers were relatively higher in built-up and agricultural areas respectively.ConclusionBacHum, BacR and Pig2Bac assays were found suitable for MST and both, human and animal faecal contaminations of drinking water sources were common in the valley.Significance and Impact of the studyMST could be an effective tool for preparing the faecal pollution strategies as these are site specific.