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首页> 外文期刊>Journal of Clinical Microbiology >Detection of Escherichia coli heat-stable enterotoxin genes in pig stool specimens by an immobilized, colorimetric, nested polymerase chain reaction.
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Detection of Escherichia coli heat-stable enterotoxin genes in pig stool specimens by an immobilized, colorimetric, nested polymerase chain reaction.

机译：通过固定的比色嵌套式聚合酶链反应检测猪粪标本中的大肠杆菌热稳定肠毒素基因。

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摘要

A combination of selective enrichment by using immunomagnetic separation of F4 (K88)-positive Escherichia coli and a nested colorimetric polymerase chain reaction (PCR) was used on crude clinical and spiked samples for determination of genes encoding heat-stable enterotoxins (STs) Ia (ST Ia) and Ib (ST Ib). The combination increased the sensitivity of the nested PCR compared with that of application onto crude samples. Dead cells were also enriched by use of this technology, giving results that are not available by traditional cultivation as enrichment before PCR. The second step in the PCR was modified to be able to differentiate between ST Ia and ST Ib genes. The colorimetric PCR was performed in a microtiter format, making it useful for automation in clinical laboratories and for the screening of large numbers of samples.
机译：通过对F4（K88）阳性大肠杆菌进行免疫磁分离和巢式比色聚合酶链反应（PCR）进行选择性富集，将其用于临床和加标样品中，以确定编码热稳定肠毒素（STs）Ia的基因ST Ia）和Ib（ST Ib）。与应用在粗样品上相比，该组合增加了巢式PCR的灵敏度。通过使用该技术还可以富集死细胞，从而获得传统培养无法在PCR之前富集的结果。修改了PCR的第二步，使其能够区分ST Ia和ST Ib基因。比色PCR以微量滴定形式进行，使其可用于临床实验室的自动化和大量样品的筛选。

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