多重定量PCR在肺孢子菌肺炎诊断中的应用

发布时间：2018-06-22 21:54

本文选题：耶氏肺孢子菌 + 肺孢子菌肺炎　；参考：《中国病原生物学杂志》2017年10期

【摘要】：目的应用靶基因的多重定量PCR方法对肺孢子菌肺炎(PCP)患者呼吸道标本中肺孢子菌进行检测,比较3种方法的检测性能。方法提取PCP患者呼吸道标本DNA,分别以Pj线粒体大亚基rRNA(mtLSUrRNA)、主要表面糖蛋白(Msg)、二氢叶酸合成酶(DHPS)为靶基因进行定量PCR检测,以临床诊断为金标准评价Mt-qPCR,Msg-qPCR和DHPS-qPCR 3种方法的敏感性、特异性、阳性预测值和阴性预测值。结果Mt-qPCR敏感性95.9%,特异性82.9%,阳性预测值83.8%,阴性预测值95.6%,Youden指数0.788,受试者工作特征曲线(ROC)下面积0.942(P0.01),95%置信区间(CI)0.909-0.975;Msg-qPCR敏感性63.9%,特异性92.4%,阳性预测值88.6%,阴性预测值73.5%,Youden指数0.563,ROC曲线下面积0.795(P0.01),95%CI 0.730-0.860;DHPS-qPCR敏感性49.5%,特异性98.1%,阳性预测值96.0%,阴性预测值67.8%,Youden指数0.476,ROC曲线下面积0.740(P0.01),95%CI0.669-0.811。PCP诊断组中Mt-qPCR、Msg-qPCR和DHPS-qPCR同时阳性43例(44.3%),任意2种方法阳性20例(占20.6%),任意1种方法阳性34例(占35.1%);非PCP组中任意2种方法阳性5例(占4.8%),任意1种方法阳性18例(占17.1%),3种方法均为阴性82例(78.1%),两组复合阳性率(100%与21.9%)比较差异有统计学意义(χ2=127.5,P0.05)。结论以临床诊断为金标准,Mt-qPCR方法优于Msg-qPCR和DHPS-qPCR,联合使用含有MtqPCR在内的2种以上qPCR方法能提高PCP诊断的符合率。
[Abstract]:Objective to detect Pneumocystis in respiratory tract samples of patients with pneumocystis pneumoniae pneumonia (PCP) by multiplex quantitative PCR with target gene. Methods DNA was extracted from respiratory tract samples of PCP patients. The sensitivity of Mt-qPCRG Msg-qPCR and DHS-qPCR was evaluated by quantitative PCR using PJ mtLSUrRNA, main surface glycoprotein (MSG) and dihydrofolate synthase (DHPS) as target genes. Specificity, positive predictive value and negative predictive value. Results the sensitivity of Mt-qPCR was 95.9, the specificity was 82.9, the positive predictive value was 83.8, the negative predictive value was 95.6and the Youden index was 0.788. The area under the operating characteristic curve (ROC) was 0.942 (P0.01) and 95% confidence interval (CI) 0.909-0.975 Msg-qPCR sensitivity (CI) was 63.9, specificity 92.4m, positive predictive value 88.6, negative predictive value 73.5 and Youden index 0.563ROC curve. Under 0.795 (P0.01), the sensitivity of CI-0.730-0.860 DHPS-qPCR is 49.5%, the specificity is 98.1, the positive predictive value is 96.0 and the negative predictive value is 67.8%. The negative predictive value is 0.740 (P0.01) ~ 95CI0.669-0.811.PCP in the diagnostic group Mt-qPCRG-qPCR and DHPS-qPCR are positive in 43 cases (44.3%), 20 cases (20.6%) are positive for any two methods, and 34 cases are positive for any one method. In the non-PCP group, 5 cases (4.8%) were positive by any two methods, and 82 cases (78.1%) were negative by any one method (17.1%). There was significant difference between the two groups (100% vs 21.9%) (蠂 2 127.5P 0.05). Conclusion Mt-qPCR based on clinical diagnosis is superior to Msg-qPCR and DHPS-qPCR. two or more qPCR methods including MtqPCR can improve the coincidence rate of PCP diagnosis.
【作者单位】：首都医科大学附属北京友谊医院北京热带医学研究所热带病防治研究北京市重点实验室;首都医科大学附属北京朝阳医院;
【基金】：首都医科大学基础临床科研合作基金项目(No.17JL03)
【分类号】：R440;R563.1

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