摘要
目的 获得四种氟喹诺酮（FQ）对结核分枝杆菌的最小抑菌浓度（MIC）,评价PCR探针熔解曲线法（PMAA）检测结核分枝杆菌对氟喹诺酮药物耐药性的临床价值.方法 124例对氧氟沙星敏感的菌株和78例对氧氟沙星耐药的菌株,在96孔细菌培养板中,用Middlebrook 7H9液体培养基将药物进行连续倍比稀释后,加入5×10^-2 mg/ml菌液100 μl,用刃天青显色法测定MIC;并用PCR探针熔解曲线法（PMAA）检测结核分枝杆菌对氟喹诺酮（FQ）药物的耐药突变.结果 氧氟沙星折点浓度为2 μg/ml,加替沙星和莫西沙星对结核分枝杆菌的MIC90比氧氟沙星和左氧氟沙星要低4-8倍,具有比氧氟沙星和左氧氟沙星更好的抗结核分枝杆菌的活性.以罗氏比例法为金标准,PCR-PMAA检测敏感度、特异度、阳性预测值、阴性预测值和符合率分别为96.2％,97.6％,96.2％,97.6％和97.0％.结论 利用微量MIC方法较常规药敏方法可以获得更多的耐药信息.用PCR-PMAA法能高效快速的检出耐药突变菌株.两种方法结合应用,对临床肺结核疾病早期用药具有更好的指导价值.
Objeciive To obtain the minimum inhibitory concentration （MIC） of four kinds of fluoroquinolones （FQ） to My cobucteriu tuberculosis and evaluate the PCR probe melting curves （PMAA） method in detection of the resistant gene for fluoroquinolones. Methods 124 ofloxacin sensitive strains and 78 ofloxacin resistance （including 48 strains of MDRTB） strains were dclt cted. Middlebrook 7H9 media containing series concentration of each drug were prepared by two fold dilu tion in the 96 well U plate. 100 bd 5 × 10^ 2mg/ml bacteria were added into each well and the MIC were determined by resazurincolorinmtric method. Fluoroquinolones （FQ） drug resistance mutations were detected by PCR probe inching curve method （PMAA） method. Results The break point concentration of ofloxacin was 2 μg/ml. The MIC, of gatifloxacin and moxifloxacin were 4- 8 times lower than that of ofloxacin and [evofloxacin,which indicates a better anti Mycobaclerium ac tivity of gatifloxacin and moxifloxacin. The detection sensitivity, specificity, positive predictive value, and negative predictive value of PCR-PMAA were 96.2 %, 97.6 %, 96.2%, 97.6% and 97 % respectively. Conclusion The MIC method can provide more valuable information about drug sensitivity than ratio method. PCR-PMAA can detect the resistance gene more quickly than traditional methods. Combination these two methods can provide more value to the treatment of TB patients.
出处
《现代检验医学杂志》
CAS
2014年第2期84-86,共3页
Journal of Modern Laboratory Medicine
基金
上海市肺科医院结核诊疗中心资助项目
关键词
结核分枝杆菌
最小抑菌浓度
探针熔解曲线法
氟喹诺酮
Mycobacterium tuberculosis
minimal inhibitory concentration
probe melting curves
{｜uoroquinolone
