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首页> 外文期刊>Applied Microbiology >New Molecular Quantitative PCR Assay for Detection of Host-Specific Bifidobacteriaceae Suitable for Microbial Source Tracking
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New Molecular Quantitative PCR Assay for Detection of Host-Specific Bifidobacteriaceae Suitable for Microbial Source Tracking

机译：用于检测适合微生物来源的宿主特异性双歧杆菌科的新分子定量PCR分析方法

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摘要

Bifidobacterium spp. belong to the commensal intestinal microbiota of warm-blooded animals. Some strains of Bifidobacterium show host specificity and have thus been proposed as host-specific targets to determine the origin of fecal pollution. Most strains have been used in microbial-source-tracking (MST) studies based on culture-dependent methods. Although some of these approaches have proved very useful, the low prevalence of culturable Bifidobacterium strains in the environment means that molecular culture-independent procedures could provide practical applications for MST. Reported here is a set of common primers and four Bifidobacterium sp. host-associated (human, cattle, pig, and poultry) probes for quantitative-PCR (qPCR) assessment of fecal source tracking. This set was tested using 25 water samples of diverse origin: urban sewage samples, wastewater from four abattoirs (porcine, bovine, and poultry), and water from a river with a low pollution load. The selected sequences showed a high degree of host specificity. There were no cross-reactions between the qPCR assays specific for each origin and samples from different fecal origins. On the basis of the findings, it was concluded that the host-specific qPCRs are sufficiently robust to be applied in environmental MST studies.
机译：双歧杆菌属属于温血动物的共生肠道菌群。一些双歧杆菌菌株显示出宿主特异性，因此被提议作为确定粪便污染起源的宿主特异性靶标。大多数菌株已用于依赖于培养物的方法的微生物来源跟踪（MST）研究中。尽管这些方法中的一些方法已被证明非常有用，但是在环境中可培养的双歧杆菌菌株的普及率较低，这意味着独立于分子培养的程序可以为MST提供实际应用。这里报道了一组通用引物和四个双歧杆菌。宿主相关（人类，牛，猪和家禽）探针，用于粪便来源跟踪的定量PCR（qPCR）评估。使用25种不同来源的水样本测试了这套水样：城市污水样本，来自四个屠宰场（猪，牛和家禽）的废水，以及来自污染负荷低的河流的水。选择的序列显示出高度的宿主特异性。每种来源特异的qPCR分析与来自不同粪便来源的样品之间没有交叉反应。基于这些发现，可以得出结论，宿主特异性qPCR具有足够的鲁棒性，可用于环境MST研究。

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