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首页> 外文期刊>Journal of Clinical Microbiology >Genetic Typing of the Porin Protein of Neisseria gonorrhoeae from Clinical Noncultured Samples for Strain Characterization and Identification of Mixed Gonococcal Infections
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Genetic Typing of the Porin Protein of Neisseria gonorrhoeae from Clinical Noncultured Samples for Strain Characterization and Identification of Mixed Gonococcal Infections

机译：从临床非培养样品中分离淋病奈瑟氏球菌的猪蛋白的基因分型，以鉴定和鉴定混合淋球菌感染。

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Molecular methods that characterize the Neisseria gonorrhoeae porin protein Por are needed to study gonococcal pathogenesis in the natural host and to classify strains from direct clinical samples used with nucleic acid amplification-based tests. We have defined the capabilities of por variable region (VR) typing and determined suitable conditions to apply the method to direct clinical specimens. Nested PCR from spiked urine samples detected 1 to 10 copies of template DNA; freezing spiked whole urine greatly reduced the ability to amplify porB. In a laboratory model of mixed gonococcal infections, the por type of one strain could be determined in the presence of a 100-fold excess of another. por VR typing was used to examine clinical samples from women enrolled in studies conducted in Baltimore, Md., and Madagascar. por type was determined from 100% of paired cervical swab and wick samples from 20 culture-positive women from Baltimore; results for eight individuals (40%) suggested infection with more than one strain. In frozen urine samples from Madagascar, porB was amplified and typed from 60 of 126 samples from ligase chain reaction (LCR)-positive women and 3 samples from LCR-negative women. The por VR types of 13 samples (21%) suggested the presence of more than one gonococcal strain. Five por types, identified in >45% of women with typed samples, were common to both geographic areas. Molecular typing is an important adjunct to nucleic acid amplification-based diagnostics. Methods that utilize direct clinical samples and can identify mixed infections may contribute significantly to studies of host immunity, gonococcal epidemiology, and pathogenesis.
机译：为了研究自然宿主中淋球菌的发病机理，并使用基于核酸扩增的试验对直接临床样本中的菌株进行分类，需要表征淋病奈瑟氏球菌孔蛋白Por的分子方法。我们定义了 por 可变区（VR）分型的功能，并确定了将该方法应用于临床标本的合适条件。从加标的尿液样品中进行的巢式PCR检测到1至10个拷贝的模板DNA。冷冻加标的全尿大大降低了 porB 的扩增能力。在混合性淋球菌感染的实验室模型中，一种 por 类型的一种菌株可以在另一种菌株过量100倍的情况下确定。 por VR分型用于检查来自马里兰州巴尔的摩和马达加斯加进行的研究的女性的临床样本。从来自巴尔的摩的20名文化阳性妇女的100％配对的宫颈拭子和灯芯样本中确定 por 类型; 8位患者（40％）的结果表明感染了一种以上的菌株。在来自马达加斯加的冷冻尿液样本中，从126个连接酶链反应（LCR）阳性女性样本和3个LCR阴性女性样本中的60个样本中扩增和鉴定了 porB 。 13种样品的 por VR类型（21％）表明存在一种以上的淋球菌菌株。这两种地区共有5种 por 类型，这些类型在> 45％的带有定型样本的女性中被发现。分子分型是基于核酸扩增的诊断方法的重要辅助手段。利用直接临床样本并可以识别混合感染的方法可能对宿主免疫，淋球菌流行病学和发病机理的研究有重大贡献。

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