New method for the visual detection of human respiratory syncytial virus using reverse transcription loop-mediated amplification

被引:10
作者
Mu, Yonglin [1 ,2 ]
Zeng, Jiawei [4 ]
Chen, Qianqian [2 ,3 ]
Liu, Jia [2 ]
Wang, Lili [2 ]
Yao, Fujia [2 ]
Cui, Meng [2 ]
He, Zhixiang [2 ]
Zhang, Chiyu [2 ]
Xiao, Ming [1 ]
Lan, Ke [2 ]
机构
[1] Shanghai Normal Univ, Coll Life & Environm Sci, Shanghai, Peoples R China
[2] Chinese Acad Sci, Inst Pasteur Shanghai, Pathogen Diagnost Ctr, Shanghai, Peoples R China
[3] Jiangsu Univ, Inst Life Sci, Zhenjiang, Jiangsu, Peoples R China
[4] Dujiangyan Med Ctr, Dujiangyan, Sichuan, Peoples R China
关键词
Human respiratory syncytial virus; RT-LAMP; Field test; Visual detection; Molecular diagnosis; ISOTHERMAL AMPLIFICATION; GENETIC-VARIABILITY; RAPID DETECTION; SUBGROUP-B; INFECTION; CHILDREN; DIAGNOSIS; ASSAY; RISK;
D O I
10.1016/j.jviromet.2014.06.005
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Human respiratory syncytial virus (HRSV) is a seasonal respiratory pathogen that causes respiratory infection in children and the elderly. Anew, reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay was developed for the rapid (within 1 h), simultaneous detection of A and B group HRSV. Primers specific for groups A and B were designed to amplify the N and L genes of HRSV, respectively. A fluorescent dye, calcein, was used as an indicator for the endpoint visual detection and/or real-time amplification of HRSV RNA:The detection limit of the new method was 281.17 50% tissue culture infective doses (TCID50)/ml for HRSV A and 1.58 TCID50/ml for HRSV B. To evaluate the validity of this method, a comparison with RT-PCR was performed using 77 nasopharyngeal swabs as samples. Both RT-LAMP and RT-PCR detected HRSV in 38 HRSV samples, yielding a positive rate of 49%. Of the RI-LAMP positive samples, 36 (95%) were also positive by RT-PCR, while two were negative by RT-PCR. Among the 36 RT-LAMP and RT-PCR positive samples, 11 belonged to HRSV group A, while 25 belonged to group B. The results show that the new RI-LAMP is simple, rapid and well suited for HRSV diagnosis, especially in a limited-resource setting. (C) 2014 Elsevier B.V. All rights reserved.
引用
收藏
页码:84 / 88
页数:5
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