半巢式聚合酶链反应扩增全血中梅毒螺旋体polA基因 |
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引用本文: | 曾铁兵,吴移谋,赵飞骏,刘双全,余敏君. 半巢式聚合酶链反应扩增全血中梅毒螺旋体polA基因[J]. 南华大学学报(医学版), 2009, 37(6): 648-650,702 |
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作者姓名: | 曾铁兵 吴移谋 赵飞骏 刘双全 余敏君 |
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作者单位: | 1. 南华大学,医学院,病原生物学研究所,湖南,衡阳,421001 2. 南华大学,第一附属医院 |
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基金项目: | 南华大学博士科研启动基金 |
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摘 要: | 目的探讨半巢式聚合酶链反应(PCR)扩增梅毒螺旋体(Tp)的DNA多聚酶Ⅰ基因(polA)以探讨检测全血标本中微量Tp的可行性。方法应用半巢式PCR和常规PCR分别扩增165例可疑梅毒及非梅毒患者全血中Tp的polA,与血清学方法比较,探讨半巢式PCR方法在扩增全血中Tp DNA的意义。结果待测的165例标本中,与血清学方法比较,半巢式PCR检测Tp的敏感性和特异性分别为62.7%和95.9%,两者符合率为82.4%;两种PCR方法检测结果差异有显著性(P〈0.01),半巢式PCR敏感性远高于常规PCR。结论半巢式polA PCR检测全血中Tp较常规PCR灵敏,是血清学方法诊断梅毒的有效补充,但其检测的灵敏度仍有待进一步提高。
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关 键 词: | 梅毒螺旋体 半巢式PCR polA 全血 |
Amplification of the DNA Polymerase I Gene of Treponema Pallidum from Whole Blood of Persons With Syphilis Using a Semi-nested PCR Assay |
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Affiliation: | ZENG Tie - bing, WU Yi - mou, ZHAO Fei - jun, et al ( Institute of Pathogenic Biology, University of South China, Hengyang, Hunan 421001, China ) |
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Abstract: | Objective To investigate the contribution of a semi - nested polA PCR assay for the detection of extremely low numbers of treponema pallidum (Tp) in whole blood of persons with syphilis. Methods Routine PCR and semi - nested PCR assays were performed to amplify specific fragments of DNA polymerase Ⅰ gene (polA) of Tp from 165 whole - blood samples of persons with suspected syphilis or non -syphilis. Compared with serology, the semi -nested PCR method was evaluated in the detection of Tp in whole blood. Results Of 165 tests performed, directly compared with serology, semi -nested PCR showed 82.4% agreement, with a sensitivity of 62.7% and a specificity of 95.9%. There were significant differences between the two PCR assays in detection of Tp from whole blood and the semi - nested polA PCR was much more sensitive than the routine polA PCR ( P 〈 0.01 ). Conclusions The semi - nestedpolA PCR assay is much more sensitive than the routine pelA PCR assay in detecting low numbers of Tp in whole blood samples as a useful addition to serology for the diagnosis of infectious syphilis. For the semi - nested polA PCR assay, the higher sensitivity for detection of Tp in whole blood should be enhanced. |
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Keywords: | polA |
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