摘要
目的探讨多色探针熔解曲线分析法(multicolor melting curve analysis,MMCA)对5种抗结核药物的耐药性诊断效果,明确MMCA检测结核分枝杆菌耐药性的临床应用价值。方法2021年4月—2022年5月选择在本院住院诊治的结核分枝杆菌阳性患者200例作为研究对象,取患者的痰液标本,分别给予传统结核分枝杆菌药敏实验(改良罗氏培养基法)和MMCA检测利福平、乙胺丁醇、链霉素、异烟肼、左氧氟沙星等5种抗结核药物的耐药性,对两种诊断方法结果不一致的样本进行基因测序验证,对比MMCA对5种抗结核药物的诊断效能。结果以结核分枝杆菌改良罗氏培养基法作为耐药诊断的金标准,MMCA检测利福平、乙胺丁醇、链霉素、异烟肼、左氧氟沙星耐药的灵敏度分别为95.83%(46/48)、93.75%(15/16)、100.00%(15/15)、100.00%(20/20)、70.00%(7/10),组间比较具有统计学差异(P<0.05),MMCA检测5种抗结核药物的特异度、阳性预测值、阴性预测值、准确度组间比较差异无统计学意义(P>0.05)。对MMCA与改良罗氏培养基法结果8例不一致的样本进行基因测序,与基因测序结果进行比较后发现,MMCA与基因测序结果符合率为75.00%(6/8)。结论MMCA在结核病患者耐药突变的检测中,对一线抗结核药物具有很高的诊断效能,对于二线抗结核药物左氧氟沙星的敏感度不足,因此在一线抗结核药物的检测上,MMCA技术具有良好的临床应用前景。
Objective To explore and analyze the diagnostic value of multicolor melting curve analysis(MMCA)for the resistance of five anti-tuberculosis drugs,so as to clarify the clinical value of MMCA in detecting drug resistance of Mycobacterium tuberculosis.Methods From April 2021 to May 2022,200 patients with positive Mycobacterium tuberculosis admitted to the Fourth People's Hospital of Qinghai Province were selected as research objects,and sputum specimens were taken from the patients.Traditional Mycobacterium tuberculosis drug sensitivity test(modified Löwenstein-Jensen medium method)and MMCA analysis were respectively given to detect the resistance of five anti-tuberculosis drugs,including isoniazid,ethambutol,streptomycin,rifampicin and isoniazid,respectively.Those samples with inconsistent results between the two diagnosis methods were subjected to gene sequencing verification,and the diagnosis efficiency of MMCA for the five anti-tuberculosis drugs was compared.Results Using Mycobacterium tuberculosis drug sensitivity as the gold standard for drug resistance diagnosis,the sensitivity of MMCA for detecting drug resistance of rifampicin,ethambutol,streptomycin,isoniazid and levofloxacin were 95.83%(46/48),93.75%(15/16),100.00%(15/15),100.00%(20/20)and 70.00%(7/10),respectively,with statistical differences between groups(P<0.05).There were no statistically significant differences in the specificity,positive predictive value,negative predictive value and accuracy of MMCA for the five anti-tuberculosis drugs(P>0.05).For the 8 samples with inconsistent results between MMCA and modified Löwenstein-Jensen medium method,gene sequencing was performed and compared with the results of gene sequencing.After comparison with gene sequencing results,it was found that the coincidence rate of MMCA and gene sequencing results was 75.00%(6/8).Conclusions In the detection of drug-resistant mutations in TB patients,multi-color probe fusion curve analysis has high diagnostic efficacy for first-line anti-tuberculosis drugs,but is not sensitive to second-line anti-tuberculosis drug levofloxacin.Therefore,for the detection of first-line anti-tuberculosis drugs,MMCA has a good clinical application prospect.
作者
常凤霞
纳元春
郝娟
朋毛措
罗莉渊
马德昭
马明
CHANG Feng-xia;NA Yuan-chun;HAO Juan;PENG Mao-cuo;LUO Li-yuan;MA De-zhao;MA Ming(Department of Clinical Laboratory,the Fourth People's Hospital of Qinghai Province,Xining,Qinghai 810000,China)
出处
《中国热带医学》
CAS
2023年第4期409-412,430,共5页
China Tropical Medicine
关键词
多色探针熔解曲线分析法
结核病
结核分枝杆菌
灵敏度
特异性
Multicolor melting curve analysis
tuberculosis
Mycobacterium tuberculosis
sensitivity
specificity
