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

Abstract

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

CLC Number:

R 519.8

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.

References

[1] HU Y, ZHANG J, LI X, et al.Penicillium marneffei infection: an emerging disease in mainland China[J]. Mycopathologia,2013,175(1-2):57-67.
[2] VANITTANAKOM N, COOPER C R, FISHER M C, et al. Penicillium marneffei infection and recent advances in the epidemiology and molecular biology aspects[J]. Clin Microbiol Rev,2006,19 (1):95-110.
[3] LE T, WOLBERS M, CHI N H, et al. Epidemiology, seasonality, and predictors of outcome of AIDS-associated Penicillium marneffei infection in Ho Chi Minh City, Viet Nam[J]. Clin Infect Dis,2011,52(7):945-952.
[4] GUO J, NING X Q, DING J Y, et al. Anti-IFN-γ autoantibodies underlie disseminated Talaromyces marneffei infections[J]. J Exp Med,2020,217(12): e20190502.
[5] CHAN J F, LAU S K, YUEN K Y, et al. Talaromyces (Penicillium) marneffei infection in non-HIV-infected patients[J]. Emerg Microbes Infect,2016,5(3): e19.
[6] LEE P P, CHAN K W, LEE T L, et al. Penicilliosis in children without HIV infection--are they immunodeficient[J]? Clin Infect Dis, 2012,54(2): e8-e19.
[7] HE L, MEI X, LU S, et al. Talaromyces marneffei infection in non-HIV-infected patients in mainland China[J]. Mycoses,2021,64(10):1170-1176.
[8] LI Y, LIN Z, SHI X, et al. Retrospective analysis of 15 cases of Penicillium marneffei infection in HIV-positive and HIV-negative patients[J]. Microb Pathog,2017,105:321-325.DOI: 10.1016/j.micpath.2017.01.026.
[9] SON V T, KHUE P M, STROBEL M.Penicilliosis and AIDS in Haiphong, Vietnam: evolution and predictive factors of death[J]. Med Mal Infect,2014,44(11-12):495-501.
[10] CHASTAIN D B, HENAO-MARTÍNEZ A F, FRANCO-PAREDES C. Opportunistic invasive mycoses in AIDS: Cryptococcosis, histoplasmosis,coccidiodomycosis, and talaromycosis[J]. Curr Infect Dis Rep,2017,19(10):36.
[11] SRINOULPRASERT Y, KONGTAWELERT P, CHAIYAROJ S C. Chondroitin sulfate B and heparin mediate adhesion of Penicillium marneffei conidia to host extracellular matrices[J]. Microb Pathog,2006,40(3):126-132.
[12] 张东伟, 蓝冰, 叶萍, 等. 马尔尼菲青霉菌病感染160例临床分析[J]. 中国医药导报, 2017, 14(32):116-119.
[13] LI X, ZHENG Y, WU F, et al. Evaluation of quantitative real-time PCR and Platelia galactomannan assays for the diagnosis of disseminated Talaromyces marneffei infection[J]. Med Mycol,2020,58(2):181-186.
[14] VANITTANAKOM N, COOPER C R JR, FISHER M C, et al. Penicillium marneffei infection and recent advances in the epidemiology and molecular biology aspects[J]. Clin Microbiol Rev,2006,19(1):95-110.
[15] XI L, LU C, ZHOU X, et al. Fifteen cases of penicilliosis in Guangdong, China[J]. Mycopathologia,2004,158(2):151-155.
[16] CHAIWUN B, KHUNAMORNPONG S, SIRIVANICHAI C, et al. Lymphadenopathy due to Penicillium marneffei infection: diagnosis by fine needle aspiration cytology[J]. Mod Pathol,2002,15(9):939-943.
[17] CAO C, LI R, WAN Z, et al. The effects of temperature, pH, and salinity on the growth and dimorphism of Penicillium marneffei[J]. Med Mycol,2007,45(5):401-407.
[18] DE PAUW B, WALSH T J, DONNELLY J P, et al. Revised definitions of invasive fungal disease from the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) Consensus Group[J]. Clin Infect Dis,2008,46(12):1813-1821.
[19] 莫冬冬. 实时荧光定量PCR诊断马尔尼菲青霉病方法的建立及其初步评价[D].南宁:广西医科大学,2014.
[20] PONGPOM M, VANITTANAKOM P, NIMMANEE P, et al. Adaptation to macrophage killing by Talaromyces marneffei[J]. Future Sci OA,2017,3(3): FSO215.
[21] KAWILA R, CHAIWARITH R, SUPPARATPINYO K. Clinical and laboratory characteristics of penicilliosis marneffei among patients with and without HIV infection in Northern Thailand: a retrospective study[J]. BMC Infect Dis,2013,13:464.DOI:10.1186/1471-2334-13-464.
[22] 谢周华,梁联哨,李志峰,等.HIV抗体阴性马尔尼菲蓝状菌病25例临床分析[J].临床肺科杂志,2019,24(9):1610-1614.
[23] CAO C, XI L, CHATURVEDI V.Talaromycosis (penicilliosis) due to Talaromyces (penicillium) marneffei: Insights into the clinical trends of a major fungal disease 60 years after the discovery of the pathogen[J]. Mycopathologia,2019,184(6):709-720.
[24] LI H R, CAI S X, CHEN Y S, et al. Comparison of Talaromyces marneffei infection in human immunodeficiency virus-positive and human immunodeficiency virus-negative patients from Fujian, China[J]. Chin Med J,2016,129(9):1059.
[25] ZHENG J, GUI X, CAO Q, et al. A clinical study of acquired immunodeficiency syndrome associated Penicillium marneffei infection from a Non-Endemic Area in China[J]. PLoS One,2015,10(6): e0130376.
[26] OSTROSKY-ZEICHNER L. Invasive mycoses: diagnosticchallenges[J]. Am J Med,2012,125(Suppl 1): S14-24.
[27] HOPE WW, WALSH T J, DENNING D W. Laboratory diagnosis of invasive aspergillosis[J]. Lancet Infect Dis,2005,5(10):609-622.
[28] MENGOLI C, CRUCIANI M, BARNES R A, et al. Use of PCR for diagnosis of invasive aspergillosis: systematic review and meta-analysis[J]. Lancet Infect Dis,2009,9(2):89-96.
[29] 郑艳青,史娜娜,曹存巍.实时荧光定量PCR检测马尔尼菲篮状菌病组织样本中菌载量的研究[J].中国真菌学杂志,2018,13(2):71-74.