title: Reverse transcription genome exponential amplification reaction assay for rapid and universal detection of human rhinoviruses.
abstract: Human rhinoviruses (HRVs) have long been recognized as the cause of more than one-half of acute viral upper respiratory illnesses, and they are associated with more-serious diseases in children, such as asthma, acute otitis media and pneumonia. A rapid and universal test for of HRV infection is in high demand. In this study, a reverse transcription genome exponential amplification reaction (RT-GEAR) assay targeting the HRV 5' untranslated region (UTR) was developed for pan-HRV detection. The reaction was performed in a single tube in one step at 65 °C for 60 min using a real-time fluorometer (Genie(®)II; Optigene). The RT-GEAR assay showed no cross-reactivity with common human enteroviruses, including HEV71, CVA16, CVA6, CVA10, CVA24, CVB5, Echo30, and PV1-3 or with other common respiratory viruses including FluA H3, FluB, PIV1-4, ADV3, RSVA, RSVB and HMPV. With in vitro-transcribed RNA containing the amplified regions of HRV-A60, HRV-B06 and HRV-C07 as templates, the sensitivity of the RT-GEAR assay was 5, 50 and 5 copies/reaction, respectively. Experiments to evaluate the clinical performance of the RT-GEAR assay were also carried out with a panel of 143 previously verified samples, and the results were compared with those obtained using a published semi-nested PCR assay followed by sequencing. The tested panel comprised 91 HRV-negative samples and 52 HRV-positive samples (18 HRV-A-positive samples, 3 HRV-B-positive samples and 31 HRV-C-positive samples). The sensitivity and specificity of the pan-HRVs RT-GEAR assay was 98.08 % and 100 %, respectively. The kappa correlation between the two methods was 0.985. The RT-GEAR assay based on a portable Genie(®)II fluorometer is a sensitive, specific and rapid assay for the universal detection of HRV infection.
authors.affiliation: Key Laboratory for Medical Virology, National Health and Family Planning Commission, Chinese Center for Disease Control and Prevention, National Institute for Viral Disease Control and Prevention, No.155 Changbai Road, Changping, Beijing, 102206, China.
authors.forename: Li
authors.initials: L
authors.lastname: Guan
authors.affiliation: Laboratory of Virology, Beijing Key Laboratory of Etiology of Viral Diseases in Children, Capital Institute of Pediatrics, 2 Yabao Road, Beijing, 100020, China.
authors.forename: Lin-Qing
authors.initials: LQ
authors.lastname: Zhao
authors.affiliation: Key Laboratory for Medical Virology, National Health and Family Planning Commission, Chinese Center for Disease Control and Prevention, National Institute for Viral Disease Control and Prevention, No.155 Changbai Road, Changping, Beijing, 102206, China.
authors.forename: Hang-Yu
authors.initials: HY
authors.lastname: Zhou
authors.affiliation: Key Laboratory for Medical Virology, National Health and Family Planning Commission, Chinese Center for Disease Control and Prevention, National Institute for Viral Disease Control and Prevention, No.155 Changbai Road, Changping, Beijing, 102206, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Nie
authors.affiliation: Key Laboratory for Medical Virology, National Health and Family Planning Commission, Chinese Center for Disease Control and Prevention, National Institute for Viral Disease Control and Prevention, No.155 Changbai Road, Changping, Beijing, 102206, China.
authors.forename: Xin-Na
authors.initials: XN
authors.lastname: Li
authors.affiliation: Key Laboratory for Medical Virology, National Health and Family Planning Commission, Chinese Center for Disease Control and Prevention, National Institute for Viral Disease Control and Prevention, No.155 Changbai Road, Changping, Beijing, 102206, China.|School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou, Guangdong, 510182, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Key Laboratory for Medical Virology, National Health and Family Planning Commission, Chinese Center for Disease Control and Prevention, National Institute for Viral Disease Control and Prevention, No.155 Changbai Road, Changping, Beijing, 102206, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Song
authors.affiliation: Laboratory of Virology, Beijing Key Laboratory of Etiology of Viral Diseases in Children, Capital Institute of Pediatrics, 2 Yabao Road, Beijing, 100020, China. yqianbjc@263.net.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Qian
authors.affiliation: Key Laboratory for Medical Virology, National Health and Family Planning Commission, Chinese Center for Disease Control and Prevention, National Institute for Viral Disease Control and Prevention, No.155 Changbai Road, Changping, Beijing, 102206, China. maxj@ivdc.chinacdc.cn.
authors.forename: Xue-Jun
authors.initials: XJ
authors.lastname: Ma
mesh_terms: D016679:Genome, Viral; D006801:Humans; D010850:Picornaviridae Infections / Q000175:diagnosis* / Q000821:virology; D012367:RNA, Viral / Q000235:genetics*; D012141:Respiratory Tract Infections / Q000175:diagnosis / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D048348:Reverse Transcription; D012229:Rhinovirus / Q000235:genetics* / Q000302:isolation & purification; D012680:Sensitivity and Specificity
chemical_list: D012367:RNA, Viral