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首页> 外文期刊>Journal of Clinical Microbiology >PCR-single-stranded confirmational polymorphism analysis for non-culture-based subtyping of meningococcal strains in clinical specimens.
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PCR-single-stranded confirmational polymorphism analysis for non-culture-based subtyping of meningococcal strains in clinical specimens.

机译：PCR-单链确证多态性分析用于临床标本中非基于脑膜炎球菌菌株的亚型分析。

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Subspecific typing of clinical meningococcal strains is important in the investigation of outbreaks and for disease surveillance. Serogrouping, typing, and subtyping of strains currently require isolation of a meningococcus from one or more clinical specimens. However, the increasing widespread practice of preadmission administration of parenteral antibiotics has resulted in a decrease in the frequency of positive cultures obtained from blood and cerebrospinal fluid. Confirmation of meningococcal disease can be obtained by meningococcus-specific PCR from both cerebrospinal fluid (H. Ni et al., Lancet 340:1432-1434, 1992) and peripheral blood (J. Newcombe et al., J. Clin. Microbiol. 34:1637-1640, 1996) specimens. However, current PCR protocols do not yield epidemiologically useful typing information. We report here the use of PCR-single-stranded confirmational polymorphism (PCR-SSCP) analysis to amplify and type meningococcal DNA present in clinical specimens. PCR-SSCP analysis with the VR1 region of the Neisseria meningitidis porA gene as the target produced unique banding patterns for each serosubtype. Direct PCR-SSCP of clinical specimens can therefore provide typing data that can be used to investigate the epidemiology of clusters of cases and outbreaks and for disease surveillance in situations in which culture of patient specimens proves negative.
机译：临床脑膜炎球菌菌株的亚型分型在暴发调查和疾病监测中很重要。菌株的血清分组，分型和亚型目前需要从一个或多个临床标本中分离出脑膜炎球菌。然而，肠胃外抗生素在入院前施用的日益普遍的实践导致从血液和脑脊髓液获得的阳性培养物的频率降低。脑膜炎球菌疾病的确认可以通过脑脊髓液（H.Ni等人，Lancet 340：1432-1434，1992）和外周血（J.Newcombe等人，J.Clin.Microbiol。 34：1637-1640，1996）标本。但是，当前的PCR协议无法产生流行病学上有用的打字信息。我们在这里报告使用PCR单链确证多态性（PCR-SSCP）分析来扩增和键入临床样本中存在的脑膜炎球菌DNA。以脑膜炎奈瑟氏菌porA基因的VR1区为靶标的PCR-SSCP分析为每种血清亚型产生了独特的条带模式。因此，临床标本的直接PCR-SSCP可以提供分型数据，这些数据可用于调查病例和暴发的流行病学，以及在患者标本培养被证明为阴性的情况下进行疾病监测。

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