优化单管巢式PCR反应条件在临床结核菌检测中的必要性

摘要/Abstract

摘要： 目的为进一步了解反应条件对PCR结果的影响?方法通过调整单管巢式PCR反应的模板量?外引物浓度和外循环数,并随机选取临床细菌学检测确定的菌阳及菌阴患者的标本进行PCR扩增检测?结果1.单管巢式PCR反应条件中的模板量及外引物循环数对检测结果的影响较大?提示在各个实验室进行PCR反应临床标本检测时,优化反应条件的重要性?2.经优化的反应条件检测临床痰标本,PCR结果与临床细菌学阳性结果的符合性较高(大于90%),对细菌学阴性患者的标本的阳性检出率为53%,并具有很高的重复性?结论证明了优化PCR反应条件的必要性?

关键词: 聚合酶链反应, 结核分枝杆菌

Abstract: Objective In order to have a better understanding of the effects of reacted conditions to PCR results.Method By adjusted the quantities of templates,concentrations of outer primers and outer cycles in one tube nested PCR.We randomly selected the positive and negative samples confirmed by clinical bacteriology and detected samples by PCR.Result (1)It is affected to the PCR results by the quantities of templates and outer cycles in one tube nested PCR.It was pointed out that it was important for the optimized condition of PCR in different laboratories.(2)By using the optimized PCR condition to detect the clinical sputum samples,the PCR results had a high correlation with the positive bacteriological samples(more than 90%).The PCR positive rate was 53% for the negative patients confirmed by bacteriology,and it could be repeated.Conclusion Optimized PCR condition was necessary when PCR as a clinical tool.

Key words: Polymerase chain reaction, M.tuberculosis

邢青, 丁北川, . 优化单管巢式PCR反应条件在临床结核菌检测中的必要性[J]. 中国防痨杂志, 1999, 21(2): 94-96.

Xing Qing,Ding Beichuan.. The necessity of optimized one tube nested PCR in clinical samples of the patients with tuberculosis[J]. Chinese Journal of Antituberculosis, 1999, 21(2): 94-96.

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