Dmitriev A, Suvorov A, Shen A D, Yang Y H
Department of Molecular Microbiology, Institute of Experimental Medicine, acad. Pavlov str., 12, St-Petersburg,197376, Russia.
Indian J Med Res. 2004 May;119 Suppl:233-6.
BACKGROUND & OBJECTIVES: The goal of the present study was to improve and simplify the diagnosis of Streptococcus agalactiae (group B Streptococcus, GBS) infection for routine clinical practice.
A total of 71 clinical samples were tested by microbiologic culture, counter immunoelectrophoresis (CIE) and PCR described in the literature. Southern hybridization was accomplished with the Enzo(TM) "DNA Labeling and Detection Kit", Roche (Germany). The computer techniques were used for selection of the specific primers and for analysis of the sizes of PCR products.
The primers for the regions around the 51 bp deletion in C5a peptidase gene (scpB) of GBS were selected. PCR analysis revealed the 255 bp amplification fragment in GBS, 306 bp fragment in groups A and G streptococci (GAS, GGS) and did not reveal any fragments in other bacterial species. Among 71 urine and serum clinical samples tested, none were found to be GBS positive by microbiologic culture, 16 samples by CIE, 36 by PCR. The specificity of amplification was confirmed by Southern hybridization.
INTERPRETATION & CONCLUSION: The 51 bp deletion in scpB gene in comparison with scpA and scpG genes can be used as a diagnostic tool for identification of GBS. The 51 bp deletion based PCR proved to be faster and more reliable test than microbiologic culture or CIE.
本研究的目的是改进并简化无乳链球菌（B族链球菌，GBS）感染的诊断方法，以用于常规临床实践。
采用文献中描述的微生物培养、对流免疫电泳（CIE）和聚合酶链反应（PCR）对71份临床样本进行检测。使用德国罗氏公司的Enzo™“DNA标记与检测试剂盒”进行Southern杂交。利用计算机技术选择特异性引物并分析PCR产物的大小。
筛选出针对GBS的C5a肽酶基因（scpB）中51bp缺失区域的引物。PCR分析显示，GBS中有255bp的扩增片段，A组和G组链球菌（GAS、GGS）中有306bp的片段，其他细菌物种中未显示任何片段。在检测的71份尿液和血清临床样本中，微生物培养未发现GBS阳性样本，CIE检测出16份阳性样本，PCR检测出36份阳性样本。通过Southern杂交证实了扩增的特异性。
与scpA和scpG基因相比，scpB基因中的51bp缺失可作为鉴定GBS的诊断工具。基于51bp缺失的PCR检测比微生物培养或CIE更快、更可靠。
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