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目的建立一种快速、敏感度高的HIV-1 DNA病毒检测方法。方法针对HIV-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-1 DNA-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-1 DNA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50 copies/ml的HIV-1 DNA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。
Objective To establish a rapid and sensitive HIV-1 DNA virus detection method. Methods Four loop-mediated isothermal amplification (LAMP) primers were designed according to the sequence of HIV-1 virus. Six rapid detection methods of HIV-1 DNA-LAMP were established and their specificity and sensitivity were evaluated. Results Peripheral lymphocytes from 200 HIV-infected patients were detected by LAMP method in 195 cases of HIV-1 DNA positive (positive rate was 97.5%), 50 healthy controls were negative. A 10-fold dilution of HIV-1 DNA virus with a minimum detectable limit of 50 copies / ml for LAMP technology was used. Hepatitis B virus, herpes virus, hepatitis C virus cross-test results were negative. Conclusion LAMP is a rapid, sensitive and specific method that is suitable for clinical examination at all levels of hospitals.