title: New method for the visual detection of human respiratory syncytial virus using reverse transcription loop-mediated amplification.
abstract: 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 1h), 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.
authors.affiliation: College of Life and Environmental Science, Shanghai Normal University, Shanghai, China; Pathogen Diagnostic Center, Institut Pasteur of Shanghai, Chinese Academy of Science, Shanghai, China.
authors.forename: Yonglin
authors.initials: Y
authors.lastname: Mu
authors.affiliation: Dujiangyan Medical Center, Dujiangyan City, Sichuan, China.
authors.forename: Jiawei
authors.initials: J
authors.lastname: Zeng
authors.affiliation: Pathogen Diagnostic Center, Institut Pasteur of Shanghai, Chinese Academy of Science, Shanghai, China; Institute of Life Sciences, Jiangsu University, Zhenjiang, Jiangsu, China.
authors.forename: Qianqian
authors.initials: Q
authors.lastname: Chen
authors.affiliation: Pathogen Diagnostic Center, Institut Pasteur of Shanghai, Chinese Academy of Science, Shanghai, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Liu
authors.affiliation: Pathogen Diagnostic Center, Institut Pasteur of Shanghai, Chinese Academy of Science, Shanghai, China.
authors.forename: Lili
authors.initials: L
authors.lastname: Wang
authors.affiliation: Pathogen Diagnostic Center, Institut Pasteur of Shanghai, Chinese Academy of Science, Shanghai, China.
authors.forename: Fujia
authors.initials: F
authors.lastname: Yao
authors.affiliation: Pathogen Diagnostic Center, Institut Pasteur of Shanghai, Chinese Academy of Science, Shanghai, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Cui
authors.affiliation: Pathogen Diagnostic Center, Institut Pasteur of Shanghai, Chinese Academy of Science, Shanghai, China.
authors.forename: Zhixiang
authors.initials: Z
authors.lastname: He
authors.affiliation: Pathogen Diagnostic Center, Institut Pasteur of Shanghai, Chinese Academy of Science, Shanghai, China.
authors.forename: Chiyu
authors.initials: C
authors.lastname: Zhang
authors.affiliation: College of Life and Environmental Science, Shanghai Normal University, Shanghai, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Xiao
authors.affiliation: Pathogen Diagnostic Center, Institut Pasteur of Shanghai, Chinese Academy of Science, Shanghai, China. Electronic address: lanke@sibs.ac.cn.
authors.forename: Ke
authors.initials: K
authors.lastname: Lan
mesh_terms: D002648:Child; D002675:Child, Preschool; D017931:DNA Primers / Q000235:genetics; D005452:Fluoresceins / Q000378:metabolism; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis* / Q000821:virology; D018113:Respiratory Syncytial Virus, Human / Q000235:genetics / Q000302:isolation & purification*; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods; D013997:Time Factors
chemical_list: D017931:DNA Primers; D005452:Fluoresceins; C007740:fluorexon
keywords: Field test; Human respiratory syncytial virus; Molecular diagnosis; RT-LAMP; Visual detection