一种基于环介导等温扩增技术原理的组织胞浆菌分子诊断技术
详细信息 查看官网全文
- 论文作者：李娟 ; 陈敏 ; 潘炜华
- 年：2016
- 作者机构：上海市医学真菌分子生物学重点实验室上海长征医院皮肤病与真菌病研究所全军皮肤病重点实验室第二军医大学附属长征医院皮肤科;
- 论文关键词：组织胞浆菌 ; 环介导等温扩增技术 ; 分子诊断 ; 快速检测
- 会议召开时间：2016-08-19
- 会议录名称：中国菌物学会2016年学术年会论文摘要集
- 英文会议录名称：Abstracts of 2016 Mycological Society of China Annual Meeting
- 语种：中文
- 分类号：R440
- 学会代码：ZGVL
- 会议名称：中国菌物学会2016年学术年会
- 会议地点：中国福建福州
- 主办单位：中国菌物学会
- 学会名称：中国菌物学会
- 页数：2
- 文件大小：587k
- 原文格式：D
- 会议级别：全国
摘要
目的研发一种基于环介导等温扩增(1oop-mediated isothermal amplification,LAMP)技术原理的组织胞浆菌感染的分子诊断的技术。方法以组织胞浆菌为研究对象,针对其种特异性的M抗原基因序列,设计并筛选数套基于LAMP技术的特异性引物,同时优化其反应条件,建立一种针对组织胞浆菌感染的分子诊断技术。结果该LAMP技术在65℃、90min反应条件下,对组织胞浆菌基因组DNA的检测敏感度达到5.3pg/反应(即5.3×10-3ng,约160个基因组DNA拷贝),且与其相近种属真菌无交叉反应。结论我们研发的这种基于LAMP技术原理的组织胞浆菌分子诊断技术具有操作简便、特异性及敏感性高等优点,具有临床推广应用的潜力。
引文
Singgh N:Fungal infections in the recipients of solid organ transplantaion[J].Infect Dis Clin North Am.2003,17:113-114
Kauffman C A.Histoplasmosis:a clinical and laboratory update[J].Clinical microbiology reviews,2007,20(1):115-132.
Craig S C.The Tropical World of Samuel Taylor Darling:Parasites,Pathology,and Philanthropy[J].Journal of the History of Medicine and Allied Sciences,2009:jrp009.
赵蓓蕾,夏锡荣,印洁,等.我国部分地区组织胞浆菌感染的流行病学调查[J].中华医学杂志(英文版),2001,第3期:199-202.
廖万清,潘搏,潘炜华.警惕在我国长江流域的组织胞浆菌病[J].皮肤性病诊疗学杂志,2014,03期(03):171-172.
Notomi T,Okayama H H,Yonekawa T,et al.Loop-mediated isothermal amplification of DNA.[J].Nucleic Acids Research,2000,28(12):E63-E63.
Tomita N,Mori Y,Kanda H,et al.Loop-mediated isothermal amplification(LAMP)of gene sequences and simple visual detection of products[J].Nature protocols,2008,3(5):877-882.
朱衡,瞿峰.利用氯化苄提取适于分子生物学分析的真菌DNA[J].菌物学报,1994,第1期(01):34-40.
Ramos-E-Silva M,Lima C M O,Schechtman R C,et al.Systemic mycoses in immunodepressed patients(AIDS).[J].Clinics in Dermatology,2012,30(6):616-627.
Baddley J W,Sankara I R,Rodriquez J M,et al.Histoplasmosis in HIV-infected patients in a southern regional medical center:poor prognosis in the era of highly active antiretroviral therapy[J].Diagnostic Microbiology&Infectious Disease,2008,62(2):151-156.
Florence H,Matthieu N,Christine A,et al.AIDS-related Histoplasma capsulatum var.capsulatum infection:25 years experience of French Guiana.[J].Aids,2008,22(9):1047-1053.
Pontes L B,Leit?o T D M J S,Lima G G,et al.Clinical and evolutionary characteristics of 134 patients with disseminated histoplasmosis associated with AIDS in the State of Ceará[J].Rev.soc.bras.med.trop,2010,43(1):27-31.
桂希恩,管立人.内脏利什曼病、播散型组织胞浆菌病及马内菲青霉菌病的诊断和鉴别[J].中国寄生虫学与寄生虫病杂志,2007,第1期(1):69-72.
Supparatpinyo K,Khamwan C,Baosoung V,et al.Disseminated Penicillium marneffei infection in southeast Asia[J].The Lancet,1994,344(8915):110-113.
Allan Jefferson G,Claudia Vera P,Herbert Leonel D M G,et al.ELISA for early diagnosis of histoplasmosis.[J].Journal of Medical Microbiology,2004,53(1):509-14.
Hung M N,Sun H Y,Hsueh P R.Meningitis due to Histoplasma capsulatum and Mycobacterium tuberculosis in a returned traveler with acquired immunodeficiency syndrome.[J].Journal of the Formosan Medical Association,2005,104(11):860-3.
Garringer T O,Wheat L J,Brizendine E J.Comparison of an established antibody sandwich method with an inhibition method of Histoplasma capsulatum antigen detection.[J].Journal of Clinical Microbiology,2000,38(8):2909-2913.
Scheel C M,Yitian Z,Theodoro R C,et al.Development of a Loop-Mediated Isothermal Amplification Method for Detection of Histoplasma capsulatum DNA in Clinical Samples.[J].Journal of Clinical Microbiology,2014,52(2):483-488.
Sara G,Cristina E,Clara V,et al.A Multiplex Real-Time PCR Assay for Identification of Pneumocystis jirovecii,Histoplasma capsulatum,and Cryptococcus neoformans/Cryptococcus gattii in Samples from AIDS Patients with Opportunistic Pneumonia[J].Journal of Clinical Microbiology,2014,52(4):1168-1176.
Hall G S,Pratt-Rippin K,,Washington J A.Evaluation of a chemiluminescent probe assay for identification of Histoplasma capsulatum isolates.[J].Journal of Clinical Microbiology,1992,30(11):3003-3004.
Chemaly R F,Tomford J W,Hall G S,et al.Rapid diagnosis of Histoplasma capsulatum endocarditis using the Accu Probe on an excised valve[J].Journal of clinical microbiology,2001,39(7):2640-2641.
Kauffman C A.Histoplasmosis:a clinical and laboratory update[J].Clinical microbiology reviews,2007,20(1):115-132.
Craig S C.The Tropical World of Samuel Taylor Darling:Parasites,Pathology,and Philanthropy[J].Journal of the History of Medicine and Allied Sciences,2009:jrp009.
赵蓓蕾,夏锡荣,印洁,等.我国部分地区组织胞浆菌感染的流行病学调查[J].中华医学杂志(英文版),2001,第3期:199-202.
廖万清,潘搏,潘炜华.警惕在我国长江流域的组织胞浆菌病[J].皮肤性病诊疗学杂志,2014,03期(03):171-172.
Notomi T,Okayama H H,Yonekawa T,et al.Loop-mediated isothermal amplification of DNA.[J].Nucleic Acids Research,2000,28(12):E63-E63.
Tomita N,Mori Y,Kanda H,et al.Loop-mediated isothermal amplification(LAMP)of gene sequences and simple visual detection of products[J].Nature protocols,2008,3(5):877-882.
朱衡,瞿峰.利用氯化苄提取适于分子生物学分析的真菌DNA[J].菌物学报,1994,第1期(01):34-40.
Ramos-E-Silva M,Lima C M O,Schechtman R C,et al.Systemic mycoses in immunodepressed patients(AIDS).[J].Clinics in Dermatology,2012,30(6):616-627.
Baddley J W,Sankara I R,Rodriquez J M,et al.Histoplasmosis in HIV-infected patients in a southern regional medical center:poor prognosis in the era of highly active antiretroviral therapy[J].Diagnostic Microbiology&Infectious Disease,2008,62(2):151-156.
Florence H,Matthieu N,Christine A,et al.AIDS-related Histoplasma capsulatum var.capsulatum infection:25 years experience of French Guiana.[J].Aids,2008,22(9):1047-1053.
Pontes L B,Leit?o T D M J S,Lima G G,et al.Clinical and evolutionary characteristics of 134 patients with disseminated histoplasmosis associated with AIDS in the State of Ceará[J].Rev.soc.bras.med.trop,2010,43(1):27-31.
桂希恩,管立人.内脏利什曼病、播散型组织胞浆菌病及马内菲青霉菌病的诊断和鉴别[J].中国寄生虫学与寄生虫病杂志,2007,第1期(1):69-72.
Supparatpinyo K,Khamwan C,Baosoung V,et al.Disseminated Penicillium marneffei infection in southeast Asia[J].The Lancet,1994,344(8915):110-113.
Allan Jefferson G,Claudia Vera P,Herbert Leonel D M G,et al.ELISA for early diagnosis of histoplasmosis.[J].Journal of Medical Microbiology,2004,53(1):509-14.
Hung M N,Sun H Y,Hsueh P R.Meningitis due to Histoplasma capsulatum and Mycobacterium tuberculosis in a returned traveler with acquired immunodeficiency syndrome.[J].Journal of the Formosan Medical Association,2005,104(11):860-3.
Garringer T O,Wheat L J,Brizendine E J.Comparison of an established antibody sandwich method with an inhibition method of Histoplasma capsulatum antigen detection.[J].Journal of Clinical Microbiology,2000,38(8):2909-2913.
Scheel C M,Yitian Z,Theodoro R C,et al.Development of a Loop-Mediated Isothermal Amplification Method for Detection of Histoplasma capsulatum DNA in Clinical Samples.[J].Journal of Clinical Microbiology,2014,52(2):483-488.
Sara G,Cristina E,Clara V,et al.A Multiplex Real-Time PCR Assay for Identification of Pneumocystis jirovecii,Histoplasma capsulatum,and Cryptococcus neoformans/Cryptococcus gattii in Samples from AIDS Patients with Opportunistic Pneumonia[J].Journal of Clinical Microbiology,2014,52(4):1168-1176.
Hall G S,Pratt-Rippin K,,Washington J A.Evaluation of a chemiluminescent probe assay for identification of Histoplasma capsulatum isolates.[J].Journal of Clinical Microbiology,1992,30(11):3003-3004.
Chemaly R F,Tomford J W,Hall G S,et al.Rapid diagnosis of Histoplasma capsulatum endocarditis using the Accu Probe on an excised valve[J].Journal of clinical microbiology,2001,39(7):2640-2641.