荧光PCR熔解曲线法对抗结核药物异烟肼、乙胺丁醇、左氧氟沙星耐药性的诊断价值

Diagnostic value of fluorescence PCR melting curve method for drug resistance of Isoniazid,Ethambutol,and Levofloxacin against tuberculosis

目的评价荧光PCR熔解曲线法对抗结核药物异烟肼、乙胺丁醇、左氧氟沙星耐药性的诊断价值。方法回顾性分析2018年1月至2019年12月首都医科大学附属北京胸科医院收治的肺结核患者170例,对表型药物敏感性试验(DST)和荧光PCR熔解曲线法对抗结核药物耐药性进行比较。以表型DST结果为金标准,分析荧光PCR熔解曲线法对抗结核药物耐药性的诊断价值。评价指标包括:敏感度、特异度、符合率、Kappa值、曲线下面积(AUC)值。同时进一步比较喹诺酮类药物(左氧氟沙星和莫西沙星)的耐药性。结果荧光PCR熔解曲线法DST敏感而表型DST耐药的例数,在异烟肼、乙胺丁醇、左氧氟沙星分别为5、5、3例。荧光PCR熔解曲线法DST耐药而表型DST敏感的例数,在异烟肼、乙胺丁醇、左氧氟沙星分别为9、16、24例。两种方法的总体不一致率,在异烟肼、乙胺丁醇、左氧氟沙星分别为8.24%、12.35%、15.88%。荧光PCR熔解曲线法检测异烟肼耐药性的敏感性、特异性、符合率、Kappa值、AUC值分别是91.53%、91.89%、91.76%、0.821、0.917;对乙胺丁醇的敏感性、特异性、符合率、Kappa值、AUC值分别是77.27%、89.19%、87.65%、0.548、0.832;对左氧氟沙星的敏感性、特异性、符合率、Kappa值、AUC值分别是89.66%、82.98%、84.12%、0.564、0.863。莫西沙星对结核杆菌的耐药率为2.94%,明显低于左氧氟沙星的17.06%(P<0.01)。结论荧光PCR溶解曲线法对异烟肼、乙胺丁醇、左氧氟沙星的耐药性有较好的辅助诊断价值。

Objective To evaluate the diagnostic value of fluorescence PCR melting curve method for drug resistance of Isoniazid,Ethambutol,and Levofloxacin against tuberculosis.Methods A retrospective analysis was performed on 170 tuberculosis patients admitted to Beijing Chest Hospital,Capital Medical University from January 2018 to December 2019.A comparison was made between phenotypic drug susceptibility testing(DST)and fluorescence PCR melting curve method for anti-tuberculosis drug resistance.The diagnostic value of fluorescence PCR melting curve method for anti-tuberculosis drug resistance was analyzed using phenotypic DST results as the gold standard.The evaluation indexes included sensitivity,specificity,coincidence rate,Kappa value,and area under the curve(AUC)value.The drug resistance of quinolones(Levofloxacin and Moxifloxacin)was further compared.Results The number of cases of DST sensitivity by fluorescence PCR melting curve method and phenotype DST resistance was 5 cases in Isoniazid,5 cases in Ethambutol,and 3 cases in Leofloxacin.The number of cases with DST resistance by fluorescence PCR melting curve method and phenotype DST sensitive was 9 cases in Isoniazid,16 cases in Ethambutol,and 24 cases in Leofloxacin.The overall inconsistent rates of Isoniazid,Ethambutol,and Levofloxacin were 8.24%,12.35%,and 15.88%,respectively.The sensitivity,specificity,coincidence rate,Kappa value,and AUC value of fluorescence PCR melting curve method to detect Isoniazid resistance were 91.53%,91.89%,91.76%,0.821,and 0.917,respectively;the sensitivity,specificity,coincidence rate,Kappa value,and AUC value of fluorescence PCR melting curve method to detect Ethambutol resistance were 77.27%,89.19%,87.65%,0.548,and 0.832,respectively;the sensitivity,specificity,coincidence rate,Kappa value,and AUC value of fluorescence PCR melting curve method to detect Levofloxacin resistance were 89.66%,82.98%,84.12%,0.564,and 0.863,respectively.The drug resistance rate of Moxifloxacin against Mycobacterium tuberculosis(2.94%)was significantly lower than that of Levofloxacin(17.06%)(P<0.01).Conclusion Fluorescence PCR melting curve method has good auxiliary diagnostic value for drug resistance of Isoniazid,Ethambutol,and Levofloxacin.