一种痰标本中结核分枝杆菌纯化浓缩方法：双膜过滤浓缩法（DMFCM）

IF 1.9 Q3 INFECTIOUS DISEASES

Journal of Clinical Tuberculosis and Other Mycobacterial Diseases Pub Date : 2025-07-04 DOI:10.1016/j.jctube.2025.100548

Dingyi Liu , Wenjing Wang , Yue Hou , Xiaoyi Gao , Wenzhao Xu , Tong Liang , Zhaogang Sun

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引用次数: 0

摘要

结核病（TB）是一种对公众健康构成全球性危害的传染病。临床部分标本中结核分枝杆菌（Mycobacterium tuberculosis， M.tb）数量少、浓度低，难以在实验室检测，影响诊断的敏感性。方法采用双膜过滤浓缩法（DMFCM）对临床痰标本中的结核分枝杆菌进行纯化浓缩，并与现有方法进行比较，评价其应用价值。结果dmfcm对80.53%的样品杂质去除率达40 ~ 80%，对mtb的富集率可达3.366倍。镜检中，与直接涂片法（DSM）相比，DMFCM检测的灵敏度由58.33%提高到73.61%。实时荧光定量PCR （qPCR）结果显示，DMFCM处理的痰标本阳性率（37/42）高于离心处理的痰标本阳性率（35/42）。在培养方面，与MGIT 960液体培养相比，DMFCM处理痰标本将确定阳性结果所需的时间减少到所需时间的75%，并提高了阳性率。结论DMFCM处理后培养液中结核分枝杆菌纯度更高，浓度更浓，可有效提高检出率。

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A method for purifying and concentrating Mycobacterium tuberculosis in sputum specimens: The double-membrane filtration and concentration method (DMFCM)

Background

Tuberculosis (TB) is an infectious disease that poses a global hazard to public health. Clinically, the small number and low concentration of Mycobacterium tuberculosis (M.tb) in some specimens make TB difficult to detect in the laboratory and affect the sensitivity of diagnosis.

Methods

M.tb in clinical sputum specimens was purified and concentrated using the double membrane filtration concentration method (DMFCM), which was compared with existing methods and evaluated for application.

Results

DMFCM removes 40–80% of impurities from 80.53% of specimens while concentrating M.tb by up to 3.366 times. In microscopy examination, compared with the direct smear method (DSM), the sensitivity with DMFCM of the test was increased from 58.33% to 73.61%. For quantitative real-time PCR (qPCR), the positive rate of the sputum specimens treated with the DMFCM (37/42) was higher than that treated with centrifugation (35/42). In terms of cultures, treatment of sputum specimens with DMFCM reduced the time required to determine a positive result to 75% of the time required compared to the MGIT 960 liquid cultures, and increased the rate of positivity.

Conclusions

The M.tb in the culture solution after DMFCM treatment is purer and more concentrated, which can effectively improve the positive rate of detection.

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来源期刊

Journal of Clinical Tuberculosis and Other Mycobacterial Diseases Medicine-Pulmonary and Respiratory Medicine

CiteScore

4.00

自引率

5.00%

发文量

审稿时长

30 weeks

期刊介绍： Journal of Clinical Tuberculosis and Mycobacterial Diseases aims to provide a forum for clinically relevant articles on all aspects of tuberculosis and other mycobacterial infections, including (but not limited to) epidemiology, clinical investigation, transmission, diagnosis, treatment, drug-resistance and public policy, and encourages the submission of clinical studies, thematic reviews and case reports. Journal of Clinical Tuberculosis and Mycobacterial Diseases is an Open Access publication.