摘要目的 比较交叉引物扩增(CPA)、恒温核酸扩增荧光检测(RealAmp)法与利福平耐药实时荧光定量核酸扩增(Xpert MTB/RIF)法对肺结核诊断的效果,评估CPA和RealAmp法在基层医院推广的价值.方法 收集2014年10月至2015年10月河南省中牟县疾病预防控制中心、新密市和邓州市结核病防治所疑诊肺结核患者3 193例,其中男2 183例,女1010例,年龄15～ 86岁,平均55岁.按照中华医学会结核病学分会制定的“肺结核诊断和治疗指南”标准诊断,确诊肺结核患者1 383例,非结核病患者1 810例.收集3份痰标本,优先选用晨痰,当无晨痰时可选择夜间痰.对3 193份痰标本涂片染色镜检,行抗酸染色及固体罗氏培养基培养,同时行CPA、RealAmp和XpertMTB/RIF法检测.对培养阳性的菌株进行MPB64抗体测定及初步菌种鉴定.以菌种鉴定后的培养结果和临床诊断结果为对照,比较CPA、RealAmp和Xpert MTB/RIF检测MTB的敏感度和特异度.结果 与固体培养结果比较,CPA、RealAmp和Xpert MTB/RIF法检测的敏感度分别为85.5％ (413/483例)、85.5％(413/483例)和87.9％(422/480例),三者比较差异无统计学意义(x2=1.6,P＞0.05);CPA、RealAmp和Xpert MTB/RIF法的特异度分别为96.8％(2 624/2 170例)、93.2％ (2 527/2 170例)和95.3％(2 567/2 170例),三者比较差异有统计学意义(x2=37.8,P＜0.001).与临床诊断结果对比,涂片镜检、固体培养、CPA、RealAmp和Xpert MTB/RIF检测MTB的敏感度分别为21.7％(300/1 383例)、34.9％(483/1 383例)、34.6％(478/1 383例)、39.2％(542/1 383例)和38.1％(526/1 381例),3种分子检测法的敏感度高于涂片镜检(x2=31.9,P＜0.01),但3种方法间的敏感度比较差异无统计学意义(x2 =2.9,P＞0.05).与临床诊断结果比较,涂片镜检、固体培养、CPA、RealAmp和Xpert MTB/RIF法的特异度分别为100％(1 810/1 810例)、100％(1 810/1810例)、98.8％(1 89/1 810例)、98.8％(1 756/1 810例)和97.0％(1 788/1 810例),3种分子检测方法间的特异度比较差异无统计学意义(x2 =0.16,P＞0.05).结论 CPA、RealAmp与XpertMTB/RIF检测痰标本的敏感度和特异度相似,但CPA与RealAmp检测快速、简便,更适于基层实验室对肺结核的诊断.

abstractsObjective To compare the efficacies of cross priming amplification CPA) and RealAmp with XpertMTB/RIF for the diagnosis of pulmonary tuberculosis (TB) at peripheral microscopic centers.Methods From December of 2014 to December of 2015,3 193 patients suspected with TB were enrolled consecutively at 3 county level TB clinical clinics in Zhongmu,Xinmi and Dengzhou of Henan province.Totally 3 193 collected sputum samples were detected by smear microscopy,L-J media culture,CPA,RealAmp and Xpert MTB/RIF.The culture positive samples were tested by MPB64 for strain identification.The sensitivity and specificity of CPA,RealAmp and Xpert MTB/RIF were calculated according to L-J solid culture results and clinical diagnosis results.Results The sensitivity of CPA,RealAmp and Xpert MTB/RIF were 85.5％ (413/483),85.5％ (413/483) and 87.9％ (422/480),respectively,compared with L-J solid culture,the difference among the 3 methods being not significant (x2 =1.6,P ＞ 0.05).The specificity of CPA,RealAmp and Xpert MTB/RIF were 96.8 ％ (2 624/2 170),93.2％ (2 527/2 170) and 95.3％ (2 567/2 170) compared with culture;and there was a significantly statistic difference among the 3 methods (x2 =37.8,P ＜ 0.001).The sensitivity of smear microscopy,culture,CPA,RealAmp and Xpert MTB/RIF was 21.7％ (300/1 383),34.9％ (483/1 383),34.6％ (478/1 383),39.2％ (542/1 383) and 38.1％ (526/1 381) compared with clinical diagnosis.The sensitivity of CPA,RealAmp and Xpert MTB/RIF was higher than that of smear (x2 =31.9,P ＜ 0.01),but there was no significantly statistic difference between the 3 molecular methods (x2 =2.9,P ＞ 0.05).The specificity of smear microscopy,L-J solid culture,CPA,RealAmp and Xpert MTB/RIF was 100％ (1 810/1 810),100％(1 810/1 810),98.8％(1 789/1 810),98.8％ (1 756/1 810) and 97.0％ (1 788/1 810),and there was no significantly statistic difference among the 3 molecular methods (x2 =0.16,P ＞ 0.05).Conclusion The capability of CPA and RealAmp for diagnosing pulmonary TB was similar to Xpert MTB/RIF.The former 2 methods were more suitable to apply to the diagnoses of pulmonary TB in peripheral laboratories.