Utility of multiplex real-time PCR in the diagnosis of extrapulmonary tuberculosis.
Utility of multiplex real-time PCR in the diagnosis of extrapulmonary tuberculosis.
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DOI：
10.1016/j.bjid.2016.01.006
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发表时间：
2016-05
影响因子：
3.4
通讯作者：
Wattal, Chand
中科院分区：
文献类型：
--
作者：
Raveendran, Reena;Wattal, Chand
肺外结核的诊断仍然是一个挑战，因为它缺乏细菌的性质。本研究的目的是评价多重聚合酶链式反应在肺外结核诊断中的作用，并比较IS6110和MPB64两种靶标检测结核分枝杆菌的效率。来自临床疑似肺结核病例的150份肺外样本(61份脓液/抽吸物，46份组织，32份体液，11份尿液)被纳入研究。所有样本都接受了直接荧光显微镜、TB培养(Bact/Alert 3D，BioMerieux，美国北卡罗来纳州达勒姆)和针对两个分枝杆菌DNA序列IS6110和MPB64的多重串联聚合酶链式反应。主混合试剂和引物由澳大利亚新南威尔士州亚历山大市AusDiagnostics Pvt.Ltd制备。采用综合金标准对检测结果进行评估，包括临床特征、微生物涂片和培养、组织病理学、细胞学、放射学和抗结核治疗的反应。涂片阳性20.3%，培养阳性23.6%，聚合酶链式反应阳性45.3%。以复合金标为标准，多重聚合酶链式反应的敏感性和特异性分别为91.9%和88.4%。其阳性预测值为85.1%，阴性预测值为93.8%。靶基因IS6110的阳性率(44.6%)高于靶基因MPB64(18.9%)。IS6110和MPB64单项指标的敏感性分别为90.3%和64.5%，特异性分别为88.4%和97.7%。聚合酶链式反应在肺外结核的快速、准确诊断中具有重要作用。在我国，仅IS6110一家就是一个有效的目标。
The diagnosis of extrapulmonary tuberculosis is still a challenge because of its pauci-bacillary nature. The aim of the study was to evaluate the role of a multiplex PCR assay in the diagnosis of extrapulmonary tuberculosis and to compare the efficiency of two targets, IS6110 and MPB64 to detect Mycobacterium tuberculosis. 150 extrapulmonary samples (61 pus/aspirate, 46 tissue, 32 body fluids, and 11 urine) from clinically suspected cases of tuberculosis were included in the study. All the samples were subjected to direct fluorescent microscopy, TB culture (BacT/ALERT 3D, biomerieux, Durham, North Carolina, USA) and a Multiplexed Tandem PCR targeting two mycobacterial DNA sequences, IS6110 and MPB64. Master-Mix reagents and primers were prepared by AusDiagnostics Pvt. Ltd (Alexandria, New South Wales, Australia). The performance of the assay was assessed using a composite gold standard, which included clinical characteristics, microbiology smear as well as culture, histopathology, cytology, radiology, and response to antitubercular therapy. 20.3%, 23.6%, and 45.3% of specimens were positive by smear, culture, and PCR, respectively. The sensitivity and specificity of the multiplex PCR was 91.9% and 88.4%, respectively, using the composite gold standard. Positive and negative predictive values of the PCR were estimated as 85.1% and 93.8%, respectively. Higher positivity was observed with target IS6110 (44.6%) as compared to target MPB64 (18.9%). The sensitivities of IS6110 and MPB64 individual targets were 90.3% and 64.5%, respectively, and specificities were 88.4% and 97.7%, respectively. PCR can play an important role in rapid and accurate diagnosis of extrapulmonary tuberculosis. IS6110 alone is an effective target in our part of the country.
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