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

Human respiratory syncytial virus (HRSV) is a seasonal respiratory pathogen that causes respiratory infection in children and the elderly. A new, reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay was developed for the rapid (within 1 h), simultaneous detection of A and B g...

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Detalles Bibliográficos
Autores principales: Mu, Yonglin, Zeng, Jiawei, Chen, Qianqian, Liu, Jia, Wang, Lili, Yao, Fujia, Cui, Meng, He, Zhixiang, Zhang, Chiyu, Xiao, Ming, Lan, Ke
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7113655/
https://www.ncbi.nlm.nih.gov/pubmed/24925133
http://dx.doi.org/10.1016/j.jviromet.2014.06.005
Descripción
Sumario:Human respiratory syncytial virus (HRSV) is a seasonal respiratory pathogen that causes respiratory infection in children and the elderly. A new, 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 RT-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 RT-LAMP is simple, rapid and well suited for HRSV diagnosis, especially in a limited-resource setting.