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首页> 外文期刊>Journal of Clinical Microbiology >Validation of nested Bordetella PCR in pertussis vaccine trial.
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Validation of nested Bordetella PCR in pertussis vaccine trial.

机译：百日咳疫苗试验中嵌套式博德氏菌PCR的验证。

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A nested PCR, using a 239-bp sequence in the pertussis toxin promoter region, was developed and evaluated. The assay differentiates Bordetella pertussis, Bordetella parapertussis, and Bordetella bronchiseptica by restriction enzyme analysis of the amplified fragments. The diagnostic performance of the PCR was evaluated in a Swedish pertussis vaccine efficacy trial which took place from 1992 to 1995, including study children and household members and using culture and serology for laboratory confirmation of suspected cases. In total 2,421 nasopharyngeal aspirates were analyzed. The total diagnostic sensitivity for B. pertussis was 90.2% (194 of 215). During the study period samples were processed with and without the cation-exchange resin Chelex. The PCR diagnostic sensitivity for B. pertussis among the Chelex-treated aspirates was 94.9% (75 of 79), and that for B. pertussis among 124 aspirates in a consecutive non-Chelex-treated material was 89.5% (111 of 124). After Chelex treatment of the 13 PCR-negative samples, an additional six became PCR positive, giving a final sensitivity of 94.3%. In addition, PCR was positive for B. pertussis with 57 of 1,744 samples negative by culture but with available serological data. The specificity of PCR with these samples was supported by a significant increase in antibody levels between acute and convalescent sera in 45 cases and by epidemiological or clinical data in all but two of the remaining cases. PCR was also positive for B. pertussis with 26 of 415 aspirates from episodes lacking serology. The diagnostic sensitivity of PCR for B. parapertussis was 74.0% (37 of 50). There were an additional seven culture-negative B. parapertussis PCR findings, six from cases with significant antibody increases against filamentous hemagglutinin only and one from a case lacking serology. There were no samples positive for B. bronchiseptica. In conclusion, PCR detection of B. pertussis and/or B. parapertussis enabled us to identify 90 positive nasopharyngeal aspirates, in addition to the 262 culture-positive samples (an increase of 34%). Relating these cases to serology and clinical data indicated a PCR specificity approaching 100%.
机译：使用百日咳毒素启动子区域的239-bp序列，开发并评估了巢式PCR。该方法通过对扩增片段进行限制性内切酶分析来区分百日咳博德氏杆菌，副百日咳博德氏杆菌和支气管败血博德特氏菌。 PCR的诊断性能在1992年至1995年的瑞典百日咳疫苗功效试验中进行了评估，该试验包括研究儿童和家庭成员，并使用培养和血清学方法对可疑病例进行实验室确认。共分析了2,421例鼻咽抽吸物。百日咳博德特氏菌的总诊断敏感性为90.2％（215中的194）。在研究期间，使用和不使用阳离子交换树脂Chelex对样品进行处理。在经过Chelex处理的抽吸物中，百日咳杆菌的PCR诊断敏感性为94.9％（79中的75），在连续的未经Chelex处理的材料中，对124抽吸物中的百日咳杆菌的PCR诊断敏感性为89.5％（124中的111）。 Chelex处理了13个PCR阴性样品后，又有6个样品成为PCR阳性，最终灵敏度为94.3％。此外，PCR对百日咳博德特氏菌呈阳性，在1,744个样品中有57个培养阴性，但有可用的血清学数据。 45例急性和恢复期血清之间抗体水平的显着提高以及其余两个病例中除其他病例外的流行病学或临床数据支持了这些样品的PCR特异性。 PCR也对百日咳博德特氏菌呈阳性，来自缺乏血清学的415例吸出物中有26例。 PCR对百日咳博德特氏菌的诊断敏感性为74.0％（50中的37）。还有另外7例培养阴性的副百日咳博德特氏菌PCR结果，其中6例来自仅针对丝状血凝素的抗体显着增加的病例，另外1例来自缺乏血清学的病例。没有支气管败血性博德氏菌阳性的样品。总之，除262个培养阳性样本外，百日咳博德特氏菌和/或百日咳博德特氏菌的PCR检测使我们能够鉴定出90个阳性鼻咽抽吸物（增加了34％）。将这些病例与血清学和临床数据相关联，表明PCR特异性接近100％。

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