结核分枝杆菌16Sr RNA+DNA和23S DNA检测对肺结核的诊断及治疗状态评价

新发传染病电子杂志 ›› 2020, Vol. 5 ›› Issue (2): 118-121.

结核分枝杆菌16Sr RNA+DNA和23S DNA检测对肺结核的诊断及治疗状态评价

邬宇美¹, 骆子义¹, 彭劲甫²

1.深圳市第三人民医院消化内科,广东深圳 518112;
2.深圳市浩鼎生物有限公司,广东深圳 518000

收稿日期:2019-10-18 出版日期:2020-02-20 发布日期:2020-06-19
通讯作者: 骆子义,Email：luoziyi630308@163.com
基金资助:
深圳市技术创新计划——创业资助项目(JCYJ20160426546)

Evaluation of Mycobacterium tuberculosis 16Sr RNA+DNA and 23S DNA detection in diagnosis and treatment of pulmonary tuberculosis

Wu Yumei¹, Luo Ziyi¹, Peng Jinfu²

1. Department of Gastroenterology1, the Third People's Hospital of Shenzhen, Guangdong Shenzhen 518112, China;
2. Shenzhen Haoding Biological Co., Ltd, Guangdong Shenzhen 518000, China

Received:2019-10-18 Online:2020-02-20 Published:2020-06-19

摘要/Abstract

摘要： 目的利用荧光定量聚合酶链反应（polymerase chain reaction assay,PCR）技术在同一体系下同时检测临床标本中结核分枝杆菌的16Sr RNA+DNA及23S DNA,同时利用其他结核分枝杆菌DNA-PCR试剂同步检测标本中结核分枝杆菌DNA以及进行结核分枝杆菌培养,探讨其不同的临床意义。方法采用多中心双盲法对来自3家医院的肺结核患者的痰标本进行结核分枝杆菌的16Sr RNA+DNA和23S DNA荧光定量PCR检测,并与结核分枝杆菌DNA荧光定量PCR检测试剂进行对比,并对标本同时进行结核分枝杆菌培养。结果 ①本研究所建立的16Sr RNA+DNA和23S DNA检测试剂与结核分枝杆菌DNA-PCR对比试剂符合率为95.76%,其中阳性符合率为95.08%,阴性符合率为96.1%。②采用本研究检测方法检测及与结核分枝杆菌培养对比,阳性符合率为81.35%,阴性符合率为87.90%,总符合率为85.92%,一致性为86%。③本研究所建立试剂盒检测结果阳性而痰培养阴性的样本共87例,其中痰培养阴性样本中凯杰试剂盒检测阳性71例;本研究所建立试剂盒检测阴性而痰培养阳性样本共58例,其中痰培养阳性样本中凯杰试剂盒检测阴性52例。④本研究检测结果与凯杰试剂盒检测结果同时阳性的样本中,将痰培养结果阳性75例与阴性34例患者的23S DNA与16Sr RNA+DNA的Ct值差值进行分析(Ct值30以上),差异有统计学意义（P<0.01）。结论同时检测16Sr RNA+DNA及23S DNA诊断结核分枝杆菌感染可能比单纯结核分枝杆菌DNA检测更灵敏,对结核分枝杆菌感染的细菌活力有一定的判断作用,对结核分枝杆菌的患者疗效评估及传染性判断有一定意义。

关键词: 结核分枝杆菌复合群, 荧光定量聚合酶链反应, 疗效评估

Abstract: Objective To detect 16Sr RNA+DNA and 23S DNA in clinical samples of Mycobacterium tuberculosis simultaneously in the same system by fluorescence quantitative PCR, and detect tuberculosis DNA in samples by other tuberculosis DNA-PCR reagents, as well as culture Mycobacterium tuberculosis. And to explore its different clinical significance. Methods Multicenter double-blinded fluorescence quantitative polymerase chain reaction (16Sr RNA+DNA/23S DNA) was used to detect sputum samples from patients with tuberculosis in three hospitals. Chi-square test and t-test were used for statistical analysis.Results The coincidence rate of 16Sr RNA+DNA/23S DNA detection reagent and tuberculosis DNA-PCR contrast reagent was 95.76%, of which the positive coincidence rate was 95.08%, and the negative coincidence rate was 96.1%. Comparing to the culture of tuberculosis bacteria, the positive coincidence rate was 81.35%, the negative coincidence rate was 87.90%, the total consistent rate was 85.92%, and the coincidence rate was 86%. There were 87 samples detected positive by our kit and sputum culture negative in this study, of which 71 were detected positive by KaiJie Kit and negative in sputum culture samples.There were 58 samples with positive sputum culture test and negative detected by our kit ,of which 52 were negative detected by KaiJie Kit. In this study, the differences of CT values between 23S DNA and 16Sr RNA+DNA of 75 positive and 34 negative sputum cultures samples were analyzed (the CT value was over 30), P<0.01. The differences were statistically significant. Conclusion 16Sr RNA + DNA detection is more sensitive than Mycobacterium tuberculosis DNA detection. 16Sr RNA+DNA/23S DNA tests has a certain role in judging the bacterial activity of Mycobacterium tuberculosis infection which is a great tool to evaluate the efficacy and infectivity of Mycobacterium tuberculosis patients.

Key words: Mycobacterium tuberculosis complex, Polymerase chain reaction assay, Therapeutic efficacy monitoring

邬宇美, 骆子义, 彭劲甫. 结核分枝杆菌16Sr RNA+DNA和23S DNA检测对肺结核的诊断及治疗状态评价[J]. 新发传染病电子杂志, 2020, 5(2): 118-121.

Wu Yumei, Luo Ziyi, Peng Jinfu. Evaluation of Mycobacterium tuberculosis 16Sr RNA+DNA and 23S DNA detection in diagnosis and treatment of pulmonary tuberculosis[J]. Electronic Journal of Emerging Infectious Diseases, 2020, 5(2): 118-121.

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