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采用表型和基因型方法对马来西亚分离出的 d-酒石酸盐阳性和 d-酒石酸盐阴性肠炎沙门氏菌亚种甲型副伤寒沙门氏菌进行鉴别。

The discrimination of d-tartrate positive and d-tartrate negative S. enterica subsp. enterica serovar Paratyphi B isolated in Malaysia by phenotypic and genotypic methods.

作者信息

Ahmad Norazah, Hoon Shirley Tang Gee, Ghani Mohamed Kamel Abd, Tee Koh Yin

机构信息

Bacteriology Unit, Institute for Medical Research, Kuala Lumpur.

出版信息

Malays J Pathol. 2012 Jun;34(1):35-9.

Abstract

Serotyping is not sufficient to differentiate between Salmonella species that cause paratyphoid fever from the strains that cause milder gastroenteritis as these organisms share the same serotype Salmonella Paratyphi B (S. Paratyphi B). Strains causing paratyphoid fever do not ferment d-tartrate and this key feature was used in this study to determine the prevalence of these strains among the collection of S. Paratyphi B strains isolated from patients in Malaysia. A total of 105 isolates of S. Paratyphi B were discriminated into d-tartrate positive (dT+) and d-tartrate negative (dT) variants by two lead acetate test protocols and multiplex PCR. The lead acetate test protocol 1 differed from protocol 2 by a lower inoculum size and different incubation conditions while the multiplex PCR utilized 2 sets of primers targeting the ATG start codon of the gene STM3356. Lead acetate protocol 1 discriminated 97.1% of the isolates as S. Paratyphi B dT+ and 2.9% as dT while test protocol 2 discriminated all the isolates as S. Paratyphi B dT+. The multiplex PCR test identified all 105 isolates as S. Paratyphi B dT+ strains. The concordance of the lead acetate test relative to that of multiplex PCR was 97.7% and 100% for protocol 1 and 2 respectively. This study showed that S. Paratyphi B dT+ is a common causative agent of gastroenteritis in Malaysia while paratyphoid fever appears to be relatively uncommon. Multiplex PCR was shown to be a simpler, more rapid and reliable method to discriminate S. Paratyphi B than the phenotypic lead acetate test.

摘要

血清分型不足以区分引起副伤寒热的沙门氏菌物种与引起较轻肠胃炎的菌株,因为这些微生物具有相同的血清型副伤寒沙门氏菌B(S. Paratyphi B)。引起副伤寒热的菌株不发酵d-酒石酸盐,本研究利用这一关键特征来确定从马来西亚患者中分离出的副伤寒沙门氏菌B菌株集合中这些菌株的流行情况。通过两种醋酸铅试验方案和多重PCR,将总共105株副伤寒沙门氏菌B分离株分为d-酒石酸盐阳性(dT+)和d-酒石酸盐阴性(dT)变体。醋酸铅试验方案1与方案2的不同之处在于接种量较低和培养条件不同,而多重PCR使用了2组靶向基因STM3356的ATG起始密码子的引物。醋酸铅方案1将97.1%的分离株鉴定为副伤寒沙门氏菌B dT+,2.9%鉴定为dT,而试验方案2将所有分离株鉴定为副伤寒沙门氏菌B dT+。多重PCR试验将所有105株分离株鉴定为副伤寒沙门氏菌B dT+菌株。醋酸铅试验相对于多重PCR的一致性,方案1和方案2分别为97.7%和100%。本研究表明,副伤寒沙门氏菌B dT+是马来西亚肠胃炎的常见病原体,而副伤寒热似乎相对不常见。与表型醋酸铅试验相比,多重PCR被证明是一种更简单、更快速且可靠的区分副伤寒沙门氏菌B的方法。

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