实时荧光定量PCR检测肺泡灌洗液在HIV阴性马尔尼菲篮状菌病诊断中的价值

中国真菌学杂志　 2023, Vol. 18 　Issue (3): 205-210.

实时荧光定量PCR检测肺泡灌洗液在HIV阴性马尔尼菲篮状菌病诊断中的价值

廖柳维^1,3, 潘开素^1,3, 李炳坤¹, 郑冬燕¹, 曹存巍^1,3, 郑艳青²

1. 广西医科大学第一附属医院皮肤科, 南宁 530021;
2. 南宁市第四人民医院传染病研究实验室, 南宁 530023;
3. 广西艾滋病防治研究重点实验室, 南宁 530021

收稿日期:2022-09-28 出版日期:2023-06-28 发布日期:2023-07-08
通讯作者: 郑艳青,E-mail:dryqzheng@yeah.net;曹存巍,E-mail:caocunwei@yeah.net E-mail:dryqzheng@yeah.net;caocunwei@yeah.net
作者简介:廖柳维,女(壮族),硕士研究生在读.E-mail:liaoliuweicn@163.com
基金资助:
广西创新研究团队(2020GXNSFGA238001);广西卫健委自筹课题(Z20210575)

Diagnostic value of quantitative real-time PCR in bronchoalveolar lavage fluid for HIV-negative talaromycosis marneffei

LIAO Liuwei^1,3, PAN Kaisu^1,3, LI Bingkun¹, ZHENG Dongyan¹, CAO Cunwei^1,3, ZHENG Yanqing²

1. Department of Dermatology, The First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China;
2. Infectious disease research laboratory, The Fourth People's Hospital of Nanning, Nanning 530023, China;
3. Guangxi Key Laboratory of AIDS Prevention and Treatment, Nanning 530021, China

Received:2022-09-28 Online:2023-06-28 Published:2023-07-08

摘要/Abstract

摘要： 目的探讨实时荧光定量PCR(quantitative real-time PCR,qPCR)技术检测HIV阴性支气管肺泡灌洗液(bronchoalveolar lavage fluid, BALF)对马尔尼菲篮状菌病诊断价值。方法收集2019年9月—2022年6月就诊于广西医科大学第一附属医院的17例HIV阴性有肺部感染症状的TSM患者及33例肺部其他真菌感染患者或非感染患者的BALF及血清标本,分别提取总DNA,运用qPCR技术检测标本中TM菌载量值,并同时进行BALF真菌培养。结果同一时间采集的17例外周血和支气管肺泡灌洗液中,在外周血样本中qPCR检测阳性仅11.8%(2/17),BALF真菌培养TM阳性患者同份BALF标本qPCR阳性率可达90.0%(9/10);BALF真菌培养TM阴性患者中,其同份标本有2例qPCR检测呈阳性。33例对照组患者的血清及BALF qPCR均为阴性。其中,BALF qPCR阳性样本的Cq值中位数为31.40,范围为20.71~35.59。同份BALF样本中,BALF qPCR技术(Kappa值:0.708)比BALF 培养(Kappa值:0.653)及BALF 镜检(Kappa值:0.220)对HIV阴性TSM患者具有更高的诊断效能。结论 qPCR技术应用于BALF标本对HIV阴性有肺部感染症状的马尔尼菲篮状菌病患者的早期诊断具有较高的应用价值。

关键词: 马尔尼菲篮状菌, 实时荧光定量PCR, HIV阴性, 支气管肺泡灌洗液, 早期诊断

Abstract: Objective To determine the loads of Talaromyces marneffei (TM) in bronchoalveolar lavage fluid (BALF) of the patients with non-HIV-infected talaromycosis marneffei (TSM) by quantitative real-time PCR (qPCR) and to evaluate the diagnostic value of it. Methods A total of 17 cases of non-HIV-infected TSM patients with pulmonary infection symptoms and 33 cases of patients with other fungal infections in the lung or non-infected patients were collected from The First Affiliated Hospital of Guangxi Medical University from September 2019 to June 2022. Samples of blood and BALF were collected from every patient at the same time. Extracting DNA from their BALF and serum, and then using qPCR to measure the loads of TM in these samples. Fungal culture of BALF was performed at the same time. Results In 17 cases of TSM, only 11.8% (2/17) of these cases were positive for qPCR measured in serum, but the positive rate of BALF qPCR was 90.0% (9/10) in patients with positive BALF fungal culture. Besides, there were 2 cases with positive BALF qPCR test results but with negative fungal culture of TM using the same sample. All the serum and BALF qPCR of other fungal infections in the lung or non-infected patients in this study were negative. In the TSM group, the median Cq value for BALF qPCR detection was 31.40 cycles (range 20.71-35.59). The qPCR showed a substantial agreement with the gold standard (kappa:0.708) and superiority to the fungal culture (kappa: 0.653) and directly microscopy-examined (kappa: 0.220). Conclusion This study showed that using qPCR to measure the loads of TM in BALF could have a high value in the early diagnosis of non-HIV-infected TSM patients with pulmonary infection symptoms.

Key words: talaromycosis marneffei (TSM), qPCR, HIV-negative, BALF, early diagnosis

中图分类号:

R 519.8

廖柳维, 潘开素, 李炳坤, 郑冬燕, 曹存巍, 郑艳青. 实时荧光定量PCR检测肺泡灌洗液在HIV阴性马尔尼菲篮状菌病诊断中的价值[J]. 中国真菌学杂志, 2023, 18(3): 205-210.

LIAO Liuwei, PAN Kaisu, LI Bingkun, ZHENG Dongyan, CAO Cunwei, ZHENG Yanqing. Diagnostic value of quantitative real-time PCR in bronchoalveolar lavage fluid for HIV-negative talaromycosis marneffei[J]. Chinese Journal of Mycology, 2023, 18(3): 205-210.

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