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Xpert 结核分枝杆菌/利福平检测、实时荧光定量聚合酶链反应和高分辨率熔解曲线在浅表淋巴结结核诊断中的应用。

The Application of Xpert Mycobacterium tuberculosis/Rifampicin, Quantitative Polymerase Chain Reaction and High Resolution Melting Curve in the Diagnosis of Superficial Lymph Node TB.

机构信息

Department of Thoracic Surgery; Beijing Chest Hospital, Capital Medical University, Beijing 101149, China.

Department of Thoracic Surgery, The Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010050, China.

出版信息

Curr Pharm Biotechnol. 2019;20(12):1044-1054. doi: 10.2174/1389201020666190716104131.

Abstract

OBJECTIVE

The diagnostic sensitivity and specificity of conventional methods for superficial lymph node tuberculosis (LNTB) are not ideal. We evaluated several novel methods including Xpert Mycobacterium tuberculosis/rifampicin (Xpert MTB/RIF) technology, quantitative fluorescence Polymerase Chain Reaction (qPCR) and High-Resolution Melting Curve (HRMC) in the diagnosis of superficial lymph node TB.

METHODS

Specimens from eighty-one consecutive patients with suspected LNTB and thirteen cases with other lymph node disease were analyzed by Xpert MTB/RIF, qPCR, and HRMC.

RESULTS

Among 81 patients with clinical suspicion of LNTB, there were 74 (91.4%) cases positive Mycobacterium tuberculosis Complex (MTBC) of Xpert MTB/RIF, 60 (74%) positive of qPCR, 24 (29.6%) of positive of BACTEC MGIT960 culture, and 13 (16%) cases positive of Roche culture. 38 cases (46.9%) were diagnosed with LNTB. All test methods showed a diagnostic specificity of 100% for LNTB. The sensitivity of molecular biology techniques was significantly higher than that of the traditional diagnostic methods, and Xpert MTB/RIF was the most sensitive diagnostic assay. On Rifampinresistant detection, Xpert MTB/RIF detected three cases (3.7%) with rpoB gene mutation, and Mycobacterium tuberculosis susceptibility testing detected 2 rifampicin-resistant cases (2.4%) which were consistent with Xpert MTB/RIF results. In the Isoniazid-resistant, 7 cases (8.1) of isoniazid resistance mutations (8.1%) were detected by HNC and 1 case was confirmed by Isoniazid susceptibility test.

CONCLUSION

Molecular detection increased the diagnostic sensitivity of LNTB and improved the detection sensitivity for rifampin and isoniazid resistance strain.

摘要

目的

传统方法诊断浅表淋巴结结核(LNTB)的诊断灵敏度和特异性不理想。我们评估了包括 Xpert 分枝杆菌/利福平(Xpert MTB/RIF)技术、定量荧光聚合酶链反应(qPCR)和高分辨率熔解曲线(HRMC)在内的几种新方法在浅表淋巴结结核诊断中的应用。

方法

对 81 例疑似浅表淋巴结结核患者和 13 例其他淋巴结疾病患者的标本进行 Xpert MTB/RIF、qPCR 和 HRMC 分析。

结果

在 81 例有 LNTB 临床疑似的患者中,Xpert MTB/RIF 阳性的结核分枝杆菌复合群(MTBC)有 74 例(91.4%),qPCR 阳性的有 60 例(74%),BACTEC MGIT960 培养阳性的有 24 例(29.6%),罗氏培养阳性的有 13 例(16%)。38 例(46.9%)诊断为 LNTB。所有检测方法对 LNTB 的诊断特异性均为 100%。分子生物学技术的灵敏度明显高于传统诊断方法,Xpert MTB/RIF 是最敏感的诊断检测方法。在利福平耐药检测方面,Xpert MTB/RIF 检测到 3 例(3.7%)rpoB 基因突变,结核分枝杆菌药敏检测到 2 例(2.4%)利福平耐药病例,与 Xpert MTB/RIF 结果一致。在异烟肼耐药方面,HNC 检测到 7 例(8.1%)异烟肼耐药突变(8.1%),1 例经异烟肼药敏试验证实。

结论

分子检测提高了 LNTB 的诊断灵敏度,提高了利福平及异烟肼耐药株的检测灵敏度。

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