A polymerase chain reaction (PCR) assay specific for Streptococcus suis based on the gene encoding the glutamate dehydrogenase
A polymerase chain reaction (PCR) assay specific for Streptococcus suis based on the gene encoding the glutamate dehydrogenase
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DOI：
10.1111/j.1574-6968.2003.tb11501.x
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发表时间：
2003-01-21
影响因子：
2.1
通讯作者：
Shull, E
中科院分区：
文献类型：
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作者：
Okwumabua, O;O'Connor, M;Shull, E
猪链球菌2型谷氨酸脱氢酶基因（gdh） 688 bp片段的聚合酶链反应（PCR）引物可以有效扩增所有306株（100%）临床猪链球菌分离株（猪305株，人1株）的DNA，包括来自不同器官和地理来源的所有精子型。当使用其他细菌的DNA作为扩增模板时，没有检测到表明引物特异性的产物。利用gdh基因引物对和猪链球菌荚膜生物合成（cps）编码基因的引物建立了多重PCR。该方法可检出血清型分别为1/2、1、2、7和9的菌株。利用多重PCR技术，先前鉴定为不可分型猪链球菌的14株（86%）中有12株（基于生化反应和血清学）的PCR结果为阳性，其中4株为血清7型阳性，3株为血清2型阳性，5株为属于其他血清型的猪链球菌。多个兽医实验室对全部14株分离株的复检结果与PCR检测结果一致，证实2株无PCR反应的分离株属于其他链球菌种类。结果表明，该方法可作为猪链球菌准确鉴定的一种可靠的种特异性分子诊断试剂，并可作为血清型特异性检测方法，分别用于血清型1/2、1、2、7和9菌株的检测。因此，PCR方法具有潜在的临床和流行病学应用价值。(C) 2002年欧洲微生物学会联合会。Elsevier Science B.V.版权所有。
Polymerase chain reaction (PCR) primers that flank a 688-bp segment within the glutamate dehydrogenase gene (gdh) of Streptococcus suis type 2 could amplify efficiently the DNA of all 306 (100%) clinical S. suis isolates tested (pigs, n = 305; human, n = 1) encompassing all scrotypes obtained from diverse organs, and geographic origins. When DNA from other bacteria were used as templates for amplification, no product was detected indicating specificity of the primers. Multiplex PCR was developed using the gdh gene primer pair and primers that targeted the gene encoding S. suis capsular biosynthesis (cps). This strategy enabled the detection of strains belonging to serotypes 1/2, 1, 2, 7, and 9, respectively. Using the multiplex-PCR technique, 12 out of 14 (86%) isolates that were previously identified as non-typable S. suis (based on biochemical reactions and serology) gave positive PCR results of which four were positive for serotype 7, three for serotype 2, and five for S. suis strains that belong to other serotypes. Retest results of all 14 isolates by several veterinary laboratories were identical with PCR and confirmed that the two non-PCR reactive isolates belonged to strains of other streptococcal species. These results indicated that PCR improved species determination and can thus be used as a reliable species-specific molecular diagnostic reagent for the accurate identification of S. suis isolates and a serotype-specific method for the detection of strains of serotypes 1/2, 1, 2, 7, and 9, respectively. The PCR method therefore has potential clinical and epidemiological applications. (C) 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.