环介导等温扩增快速检测HIV一1DNA病毒 |
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引用本文: | 张永乐,潘克女,章松平,朱明利,蔡兆斌,刘寿荣,厉小玉,周俊. 环介导等温扩增快速检测HIV一1DNA病毒[J]. 实用全科医学, 2014, 0(4): 615-616 |
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作者姓名: | 张永乐 潘克女 章松平 朱明利 蔡兆斌 刘寿荣 厉小玉 周俊 |
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作者单位: | [1]浙江省杭州市西溪医院分子诊断实验室,310000 [2]浙江省杭州市西溪医院检验科 ,310000 [3]浙江省杭州市西溪医院感染科 ,310000 [4]杭州市儿童医院检验科,310000 |
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基金项目: | 杭州市科技局重点计划项目(20110733Q06) |
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摘 要: | 目的建立一种快速、敏感度高的HIV.1DNA病毒检测方法。方法针对HIV一1型病毒序列的特点寻找6个相对保守的区域设计4条环介导等温扩增(LAMP)引物,建立HIV一1DNA—LAMP快速检测方法,并评价其特异性和灵敏度。结果200例HIV感染者外周淋巴细胞中,应用LAMP法检测出195例HIV一1DNA阳性(阳性率为97.5%),50例健康对照结果阴性。采用10倍稀释法,LAMP技术的最低检出限为50copies/m1的HIV一1DNA病毒。对乙型肝炎病毒、疱疹病毒、丙型肝炎病毒进行交叉实验结果均为阴性。结论LAMP是一种快速、敏感度高、特异性强的方法,适合各级医院的临床检测。
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关 键 词: | 环介导等温扩增 艾滋病病毒 快速诊断 |
Development and evaluation of loop-mediated isothermal amplification for rapid detection of human immunodefi- ciency virus type-1 |
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Affiliation: | DNA ZHANG Yong-le, L[ Xiao-yu, PAN Ke-nv, et al. Molecular Diagnosis Laboratory, Hangzhou Xixi Hospital, Hangzhou 310023, Zhejiang , Chin |
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Abstract: | Objective The objective of this study was to establish a loop-mediated isothermal amplification (LAMP) method for rapid ancl effective detection of human immunodeficiency virus type 1 ( HIV-1 ) DNA. Methods Four LAMP primers specific to six regions of the a conserved part of the HIV-1 genome were designed. We established a novel LAMP method for detecting HIV-1 DNA isolates among 200 HIV-l-infected individuals. Results The LAMP assay detected HIV-1 DNA in peripheral lymphocytes in 195 of 200 samples from 200 HIV-l-infected individuals. All of the control spec- imens showed negative. The detection sensitivity of the method was found to be 50 copies/ml. Using this LAMP assay, the results of cross experimental including Hepatitis B virus, herpes virus, hepatitis C virus, were negative. Conclusion Al- most all HIV-DNA remained in peripheral blood of HIV infection. This LAMP method was convenient, rapid and accurate in detecting HIV-1 DNA. It appeared to be useful for routine clinical practice. |
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Keywords: | Loop-Mediated isothermal amplification (LAMP) HIV-RNA Rapid detection |
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