分子线性探针技术在检测脊柱标本中结核分枝杆菌及其耐药性中的应用价值

The value of linear probe assay(HAIN) for the diagnosis of M. tuberculosis and the detection of rifampin and isoniazid resistance in spinal tuberculosis

中文关键词:  脊柱结核  分子线性探针技术  药敏试验  耐药性

英文关键词:Mycobacterium tuberculosis  Spine  Drug sensitivity testing  Drug resistance  Linear probe assay

中文摘要:

  【摘要】 目的：评估分子线性探针技术（HAIN）应用于检测脊柱标本中的结核分枝杆菌（MTB）及其耐药性的临床价值。方法：回顾性分析首都医科大学附属北京胸科医院骨科2015年6月~2016年12月收治的脊柱感染的患者235例，其中男132例，女103例，年龄1～84岁，平均42.8±18.1岁，根据伽马干扰素释放试验、标本普通细菌培养和真菌培养结果进行分组，脊柱结核（结核组）219例，脊柱感染其他细菌或真菌（非结核组）16例，通过穿刺或手术获取脓液或组织标本（含肉芽组织、干酪样坏死物和骨质），然后在2h内采用HAIN技术和联合培养法对相同标本进行MTB检测，并进行抗结核药物的敏感性试验。比较两种方法检测MTB的敏感度和特异度，以及检测利福平耐药、异烟肼耐药和耐多药的敏感度和特异度。结果：HAIN和联合培养法检测MTB结果为阳性的分别有120例和104例。HAIN和联合培养法检测MTB的敏感度分别为54.8%和47.5%，特异度均为100%。HAIN在MTB培养阳性标本和培养阴性标本中的阳性率分别为63.5%、47.0%，两组的阳性率的差异有统计学意义（χ2=6.006，P<0.05）。HAIN检测利福平耐药、异烟肼耐药和耐多药的敏感度分别为87.5%、66.7%和85.7%，特异度分别为94.8%、86.0%和93.2%，一致性检验Kappa值分别为0.743（P<0.001）、0.427（P<0.001）和0.664（P<0.001）。结论：HAIN在检测脊柱结核中的结核分枝杆菌及其利福平耐药和异烟肼耐药中展现出了高敏感度和特异度的技术特点，具有良好的临床应用前景。

英文摘要:

  【Abstract】 Objectives: To evaluate the value of Hain GenoType MTBDR plus(HAIN) for the diagnosis of M. tuberculosis and the detection of rifampin(RIF) and isoniazid(INH) resistance in spinal tuberculosis. Methods: 235 hospitalized patients with spinal infection were enrolled, including 132 males and 103 females, aged from 1 to 84 years with an average age of 42.8±18.1 years, selected from the Orthopedic Department of Beijing Chest Hospital affiliated to Capital Medical University from June 2015 to December 2016. According to the results of interferon gamma release test, common bacterial culture and fungal culture,all patients were divided into two groups. One was tuberculous group including 219 cases with spinal tuberculosis and the other group was non-tuberculous group including 16 cases with spinal infection with non-tuberculous bacterium or fungus. All patients′ specimens were collected using needle puncture or surgery and sent for the detection of MTB using HAIN and co-culture within 2 hours. The co-culture was to carry out BACTEC MGIT 960 system culture and modified Roche medium culture at the same time. As long as one method of culture was to be positive, co-culture was to be identified as positive. Then the culture positive strains were to be performed for drug sensitivity test(DST). The sensitivity and specificity of HAIN for detection of MTB was to be compared with co-culture. The sensitivity and specificity of detection of RIF resistance and INH resistance and multiple drug-resistance(MDR, both RIF and INH resistance) were to be compared between HAIN and DST. The consistency of HAIN test results was evaluated according to the results of DST. Results: The positive detection of MTB was found in 120 cases and 104 cases using HAIN and co-culture, respectively. The sensitivity of detection of MTB was 54.8% and 47.5% without a significant difference(χ2=2.339，P=0.126), respectively. The specificity using HAIN and co-culture were both 100%. The positive rate of detection of MTB was 63.5% and 47.0%, respectively, in culture positive specimens and culture negative specimens using HAIN, with a significant difference (χ2=6.006, P<0.05). Compared with the DST, the sensitivity and specificity of detection of RIF resistance, INH resistance and MDR was 87.5% and 94.8%(Kappa=0.743, P<0.001), 66.7% and 86.0%(Kappa=0.472, P<0.001), 85.7% and 93.2%(Kappa=0.664, P<0.001), respectively, using HAIN. Conclusions: The HAIN, which is an accurate tool for the detection of MTB and its RIF resistance and INH resistance, has a good application prospect in spinal tuberculosis．