通过固定化比色巢式聚合酶链反应检测猪粪便标本中的大肠杆菌热稳定肠毒素基因
Detection of Escherichia coli heat-stable enterotoxin genes in pig stool specimens by an immobilized, colorimetric, nested polymerase chain reaction.
作者信息
Hornes E, Wasteson Y, Olsvik O
机构信息
Department of Microbiology and Immunology, Norwegian College of Veterinary Medicine, Oslo.
出版信息
J Clin Microbiol. 1991 Nov;29(11):2375-9. doi: 10.1128/jcm.29.11.2375-2379.1991.
Abstract
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.
摘要
采用免疫磁珠分离法对F4（K88）阳性大肠杆菌进行选择性富集，并结合巢式比色聚合酶链反应（PCR），用于检测临床原始样本和加标样本中编码热稳定肠毒素（ST）Ia（ST Ia）和Ib（ST Ib）的基因。与直接应用于原始样本相比，这种方法提高了巢式PCR的灵敏度。利用该技术还能富集死细胞，得到传统培养法（作为PCR前的富集方法）无法获得的结果。对PCR的第二步进行了改进，以便能够区分ST Ia和ST Ib基因。比色PCR以微孔板形式进行，适用于临床实验室的自动化操作和大量样本的筛查。
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