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支气管肺泡灌洗液中检测肺结核的分子诊断方法与传统诊断方法的评估及比较

Evaluation and comparison of molecular and conventional diagnostic modalities for detecting pulmonary tuberculosis in bronchoalveolar lavage fluid.

作者信息

Bhatia Disha, Bhatia Nirmaljit Kaur, Deepak Desh, Sharma Brijesh, Shulania Anuradha, Duggal Nandini

机构信息

Dr. Ram Manohar Lohia Hospital and PGIMER, New Delhi, 110001, India.

出版信息

Indian J Med Microbiol. 2021 Jan;39(1):48-53. doi: 10.1016/j.ijmmb.2020.10.003. Epub 2020 Nov 5.

Abstract

CONTEXT

In cases of sputum smear-negative and sputum-scarce (SSN/SC) pulmonary tuberculosis (PTB), bronchoalveolar lavage (BAL) fluid may be helpful in establishing diagnosis. No specific recommendations for BAL samples have yet been formulated due to limited literature.

AIMS

  1. To find a sensitive and specific protocol for same-day diagnosis of PTB using BAL in SSN/SC clinically suspected patients. 2. To evaluate the need to routinely perform MGIT for all BAL samples.

SETTINGS AND DESIGN

Prospective observational study design in a tertiary care hospital in New Delhi.

METHODS AND MATERIAL

Fibreoptic bronchoscopy was performed and BAL collected from 175 clinically suspected SSN/SC PTB patients. BAL samples were subjected to: ZN Stain, Xpert MTB/RIF CBNAAT, BACTEC MGIT 960 liquid culture and M. tuberculosis complex DNA Real time PCR. The results of the various diagnostic tests were analysed using a) MGIT as gold standard and b) a composite reference standard (CRS) for a final diagnosis of PTB.

STATISTICAL ANALYSIS USED

Microsoft Excel 2016 and SPSS version 21.0 were used. Sensitivity, specificity and predictive values were calculated and compared using McNemar test. A p value of <0.05 was considered statistically significant.

RESULTS

34 Cases had a final diagnosis of TB as per the CRS. Using CRS, MGIT had a sensitivity of 50.0% (32.4%-67.6%). There was no statistically significant difference between sensitivities of CBNAAT and PCR; both were more sensitive than ZN stain. Sensitivity and specificity of CBNAAT was 79.4% (62.1%-91.3%) and 100.0% (97.4%-100.0%) respectively. The preferred protocol for the hospital is CBNAAT and ZN stain. There was no statistically significant difference in sensitivity by adding PCR or MGIT to this protocol.

CONCLUSIONS

We found it a good strategy to perform CBNAAT and ZN stain on BAL fluid for accurate and same-day PTB diagnosis. CBNAAT is useful for ruling PTB in even when BAL cultures are negative. It is prudent to continue to routinely perform MGIT for all BAL samples.

摘要

背景

在痰涂片阴性和痰量稀少(SSN/SC)的肺结核(PTB)病例中,支气管肺泡灌洗(BAL)液可能有助于确诊。由于文献有限,尚未制定针对BAL样本的具体建议。

目的

  1. 为临床疑似SSN/SC的患者找到一种使用BAL进行PTB当日诊断的敏感且特异的方案。2. 评估对所有BAL样本常规进行分枝杆菌生长指示管(MGIT)检测的必要性。

设置与设计

在新德里一家三级护理医院进行前瞻性观察性研究设计。

方法与材料

对175例临床疑似SSN/SC PTB患者进行纤维支气管镜检查并收集BAL样本。BAL样本进行以下检测:萋尼染色、Xpert MTB/RIF环介导等温扩增技术(CBNAAT)、BACTEC MGIT 960液体培养以及结核分枝杆菌复合群DNA实时聚合酶链反应(PCR)。使用以下两种方法分析各种诊断检测的结果:a)以MGIT作为金标准;b)采用综合参考标准(CRS)进行PTB的最终诊断。

所用统计分析方法

使用Microsoft Excel 2016和SPSS 21.0版本。计算敏感性、特异性和预测值,并使用McNemar检验进行比较。p值<0.05被认为具有统计学意义。

结果

根据CRS,34例最终诊断为结核病。以CRS为标准,MGIT的敏感性为50.0%(32.4%-67.6%)。CBNAAT和PCR的敏感性之间无统计学显著差异;两者均比萋尼染色更敏感。CBNAAT的敏感性和特异性分别为79.4%(62.1%-91.3%)和100.0%(97.4%-100.0%)。该医院首选的方案是CBNAAT和萋尼染色。在此方案中添加PCR或MGIT,敏感性无统计学显著差异。

结论

我们发现对BAL液进行CBNAAT和萋尼染色是准确进行PTB当日诊断的良好策略。即使BAL培养结果为阴性,CBNAAT也有助于确诊PTB。对所有BAL样本继续常规进行MGIT检测是谨慎的做法。

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