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一种快速双链实时PCR检测方法直接从培养平板对空肠弯曲菌和结肠弯曲菌进行菌种鉴定的适用性。

Applicability of a rapid duplex real-time PCR assay for speciation of Campylobacter jejuni and Campylobacter coli directly from culture plates.

作者信息

Best Emma L, Powell Ella J, Swift Craig, Grant Kathleen A, Frost Jennifer A

机构信息

Campylobacter Reference Unit, Laboratory of Enteric Pathogens, Specialist and Reference Microbiology Division, Health Protection Agency, 61, London, NW9 5HT, UK.

出版信息

FEMS Microbiol Lett. 2003 Dec 12;229(2):237-41. doi: 10.1016/S0378-1097(03)00845-0.

Abstract

A rapid duplex real-time polymerase chain reaction (PCR) assay for speciation of Campylobacter jejuni and Campylobacter coli using the ABI Prism 7700 sequence detection system (Applied Biosystems) was developed based on two of the genes used in a conventional multiplex PCR. A rapid turnaround time of 3 h was achieved with the use of boiled cell lysates. Applicability of the assay was tested with 6015 random campylobacter strains referred to the Campylobacter Reference Unit, with 97.6% being identified as either C. jejuni or C. coli by this technique. Rapidity, combined with specificity and sensitivity, makes this method for routine campylobacter speciation attractive to any laboratory with a Taqman system.

摘要

基于传统多重聚合酶链反应(PCR)中使用的两个基因,开发了一种利用ABI Prism 7700序列检测系统(应用生物系统公司)对空肠弯曲菌和结肠弯曲菌进行菌种鉴定的快速双重实时聚合酶链反应(PCR)检测方法。使用煮沸的细胞裂解物可实现3小时的快速周转时间。用提交给弯曲菌参考单位的6015株随机弯曲菌菌株对该检测方法的适用性进行了测试,通过该技术,97.6%的菌株被鉴定为空肠弯曲菌或结肠弯曲菌。该方法的快速性、特异性和敏感性相结合,使得这种用于常规弯曲菌菌种鉴定的方法对任何拥有Taqman系统的实验室都具有吸引力。

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