首页|荧光PCR熔解曲线法检测结核分枝杆菌对氟喹诺酮类耐药性的临床研究

荧光PCR熔解曲线法检测结核分枝杆菌对氟喹诺酮类耐药性的临床研究

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目的 评估荧光PCR熔解曲线法早期诊断结核病患者对氟喹诺酮类(FQs)耐药性的效能,分析FQs耐药情况及特征,为利福平耐药/耐多药结核病(RR/MDR-TB)、准广泛耐药结核病(pre-XDR-TB)规范诊断和选择合理治疗方案提供依据。方法 收集2021年1月至2022年8月贵阳市公共卫生救治中心1094例门诊和住院患者涂片阳性标本,行罗氏固体培养法、菌种鉴定,最终589例结核病患者均进行表型药敏试验与荧光PCR熔解曲线法,对利福平(RFP)、异烟肼(INH)、乙胺丁醇(EMB)、FQs耐药性进行检测;以表型药敏试验为标准,评估荧光PCR熔解曲线法的诊断效能,根据表型药敏试验结果分析患者FQs耐药情况和临床特征的关系。结果 荧光PCR熔解曲线法检测结核病患者对FQs耐药性的灵敏度、特异度、符合率、Kappa值分别为91。30%、97。69%、96。94%、0。86,曲线下面积(AUC)为0。945,高于RFP、INH、EMB的0。924、0。923、0。850。RR/MDR-TB患者中FQs耐药率为22。80%,荧光PCR熔解曲线法检测患者对FQs耐药性的Kappa值为0。83,一致性较好,AUC为0。936。荧光PCR熔解曲线法检测不同细菌载量结核病患者对FQs耐药性的灵敏度、特异度、符合率差异均无统计学意义(P>0。05)。治疗类型、抗结核史、肺部空洞、MDR-TB与FQs耐药有关(P<0。05)。结论 荧光PCR熔解曲线法对结核病患者FQs耐药性有较好的诊断效能。
Clinical study on fluoroquinolones resistance of Mycobacterium tuberculosis detected by fluorescence PCR melting curve method
Objective To evaluate the efficiency of fluorescent PCR melting curve method in early diag-nosis of fluoroquinolones (FQs) resistance in the patients with tuberculosis,and to analyze the situation and characteristics of FQs resistance,so as to provide a basis for the standardized diagnosis and treatment of rifam-picin resistance/multidrug resistant tuberculosis (RR/MDR-TB) and pre-extensively drug resistant tuberculo-sis (pre-XDR-TB).Methods A total of 1094 smear positive samples from the outpatients and inpatients of Guiyang Municipal Public Health Treatment Center from January 2021 to August 2022 were collected and conducted the Roche solid culture method and bacterial species identification.Finally,589 cases of tuberculosis conducted the phenotypic drug sensitivity test and fluorescent PCR melting curve method for detecting rifam-picin (RFP),isoniazid(INH),ethambutol(EMB) and FQs resistance.The phenotypic drug sensitivity test served as the standard to evaluated the diagnostic efficiency of the fluorescent PCR melting curve method;the relationship between the patients' FQs resistance and clinical characteristics was analyzed according to the phenotypic drug sensitivity results.Results The sensitivity,specificity,coincidence rate and Kappa value of fluorescence PCR melting curve method for detecting FQs drug resistance were 91.30%,97.69%,96.94% and 0.86 respectively;the area under the curve (AUC) was 0.945,which was higher than 0.924,0.923 and 0.850 of RFP,INH and EMB.The drug resistance rate of FQs in the patients with RR/MDR-TB was 22.80%,the Kappa value of fluorescence PCR melting curve method for detecting the patients' FQs drug re-sistance was 0.83,the consistency was good,AUC was 0.936.There was no statistically significant difference in sensitivity,specificity and coincidence rate of FQs resistance in TB patients with different bacterial loads by fluorescence PCR fusion curve (P>0.05).The treatment type,history of anti-tuberculosis,pulmonary cavity and MDR-TB were related with FQs resistance (P<0.05).Conclusion The fluorescent PCR melting curve method has good diagnostic efficiency for FQs resistance in the patients with tuberculosis.

mycobacterium tuberculosisdrug resistancefluorescence PCR melting curve methodfluoroquinolonespre-extensively drug resistant tuberculosis

秦娅莉、陈静、李军、王明栋、欧维正、邱继瑶、彭燕清

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贵州医科大学公共卫生与健康学院环境污染与疾病监控教育部重点实验室,贵阳 561113

贵阳市公共卫生救治中心,贵阳 550001

耐多药结核病 耐药性 荧光PCR熔解曲线法 氟喹诺酮类 准广泛耐药结核病

"十三五"国家重大新药创制项目

2017ZX09304009

2024

重庆医学
重庆市卫生信息中心,重庆市医学会

重庆医学

CSTPCD
影响因子:1.797
ISSN:1671-8348
年,卷(期):2024.53(16)