环介导等温扩增法在登革病毒临床检测中的运用

摘要

Objective To investigate the feasibility of loop-mediated isothermal amplification (LAMP) applied in the rapid diagnosis of dengue virus.Methods Peripheral blood samples were collected from 226 dengue fever patients and 30 healthy subjects during physical examination.After serum isolation,the loading capacity of dengue virus RNA was measured by serological antibody-specific method,LAMP and realtime quantitative PCR,respectively.The detection results among three approaches were statistically compared.Results Among 226 serum samples,the positive rate was 84.5％ detected by serological antibody-specific method,95.6％ for real-time quantitative PCR and 96.9％ for LAMP.Both real-time quantitative PCR and LAMP mainly detected type Ⅰ dengue virus.Negative results of dengue virus were obtained in the healthy individuals by three techniques.Within the range of 104-108 copies of dengue virus,the reaction time of LAMP was shortened along with the increasing loading capacity of dengue virus.Conclusion LAMP can be utilized for rapid detection of dengue virus.%目的 探讨利用环介导等温扩增(LAMP)快速诊断登革病毒的可行性.方法 收集226例登革热患者和30名健康体检者的外周血标本,分离血清,分别使用血清特异性抗体、LAMP及实时定量PCR检测登革病毒RNA载量,统计分析各种检测方法应用情况.结果 226例登革热患者血清样本中,登革病毒血清抗体阳性率为84.5％,PCR检测登革病毒RNA阳性率为95.6％,LAMP检测登革病毒阳性率96.9％,PCR及LAMP所检测的登革病毒血清分型均以1型为主.3种方法对30名健康体检者的登革病毒检测结果均为阴性.在登革病毒104 ～ 108 copies的范围内,随着病毒载量的增加,LAMP的反应时间随之缩短.结论 LAMP可用于登革病毒的快速检测.