荧光PCR溶解曲线法在耐药肺结核快速诊断中临床价值的探讨  

Clinical value of rapid diagnosis of drug-resistant tuberculosis by fluorescence PCR dissolution curve

作　　者：邹远妩 王婷[2] 王智存 梁亚萍 王卓 屈少毅 李静 ZOU Yuan- wu;WANG Ting;WANG Zhi-cun;LIANG Ya-ping;WANG Zhuo;QU Shao-yi;LI Jing(Department of Laboratory,Tuberculosis Control Hospital of Shaanxi Province,Xi'an,Shaanxi710100,China;Department of Nuclear Medicine,Ankang Central Hospital,Ankong,Shannxi725000,China)

机构地区：[1]陕西省结核病防治院检验科,陕西西安710100 [2]安康市中心医院核医学科,陕西安康725000

出　　处：《临床肺科杂志》2019年第10期1882-1885,共4页Journal of Clinical Pulmonary Medicine

基　　金：陕西省扶助项目(No 2014D10)

摘　　要：目的评价荧光PCR溶解曲线法(溶解曲线法)检测结核分枝杆菌耐药突变的效果。方法用液体传统药敏与溶解曲线法同时检测121株结核分枝杆菌临床分离株对链霉素(Streptomycin,SM)、异烟肼(Isoniazid,INH)、利福平(Rifampicin,RFP)、乙胺丁醇(Ethambutol,EMB)4种一线抗结核药物的耐药突变,罗氏传统药敏检测菌株对左氧氟沙星(Levofloxacin,LFX)和莫西沙星(MoxifLoxacin,MFX)的耐药性与溶解曲线法检测菌株氟喹诺酮类耐药突变,并进行比较。结果传统药敏与溶解曲线法检测,5种抗结核药物耐药性差异无统计学意义(P>0.05),而氟喹诺酮类中莫西沙星耐药性差异有统计学意义(P<0.05)。同一菌株,以传统药敏法为标准,溶解曲线法检测SM、INH、RFP、EMB、氟喹诺酮类中左氧氟沙星耐药的敏感度分别为84.84%、72.73%、93.75%、80.00%、92.31%,特异度分别为93.18%、98.86%、97.75%、93.40%、90.74%,两种方检测法的一致性分析Kappa值分别为0.7803、0.7159、0.9150、0.7340、0.8305,表明两者一致性好;两种方法对氟喹诺酮类中莫西沙星(MFX)耐药检测敏感度、特异度、约登指数分别为21.74%,99.01%,0.2075,表明两种方法一致性差。结论熔解曲线法与传统法对SM、INH、RFP、EMB、LFX耐药性检测效果一致,且快速,敏感度、特异度较高,可明显缩短结核诊疗时间;而熔解曲线法对MFX检测效果一般,敏感性、一致性较低。溶解曲线法在耐药结核病诊断、治疗上有较好指导意义。Objective To evaluate the efficacy of detection of drug-resistant mutations in mycobacterium tuberculosis by fluorescence PCR.Methods It used traditional liquid drug susceptibility and dissolution curve method to test the resistant mutation of 121 strains of mycobacterium tuberculosis clinical isolates to streptomycin(SM),isoniazid(INH),rifampicin(RFP)and ethambutol(EMB)four first-line anti-TB drugs.The results of Roche traditional drug susceptibility test for moxifloxacin and levofloxacin resistance strains and dissolution curve method fluoroquinolone resistance mutation were compared.Results According to the traditional drug sensitivity and dissolution curve test,there was no statistically significant difference in drug resistance of the five anti-tuberculosis drugs(P>0.05),while there was statistically significant difference in drug resistance of moxifloxacin among fluoroquinolones(P<0.05).The same strain,on the basis of traditional drug susceptibility method,the sensitivity of SM,INH,RFP,EMB,and fluoroquinolone centre-left ofloxacin(LFX)were 84.84%,72.73%,93.75%,80.00%,92.31%,and their specificity were 93.18%,98.86%,97.75%,93.40%,and 90.74%respectively for dissolve curve method.The consistency of the two method Kappa value were 0.7803,0.7159,0.9150,0.7340,and 0.8305,which showed that the consistency between them was good.The sensitivity,specificity and approximate indexes of moxifloxacin(MFX)resistance in fluoroquinolones detected by the two methods were 21.74%,99.01%and 0.2075,respectively,indicating poor consistency of the two methods.Conclusion The dissolution curve method has the same effect as the traditional method in detecting the drug resistance of SM,INH,RFP,EMB and levofloxacin,and it is rapid,sensitive and specific,which can significantly shorten the time of diagnosis and treatment of tuberculosis.However,the fusion curve method has a general effect on moxifloxacin detection,with low sensitivity and consistency.The dissolution curve method has a good guiding significance in the diagnosis and treatm

关 键 词：结核分枝杆菌 溶解曲线法 耐药突变 耐药结核 

分 类 号：R521[医药卫生—内科学]