title: Identification of human metapneumovirus genotypes A and B from clinical specimens by reverse transcription loop-mediated isothermal amplification.
abstract: Human metapneumovirus (hMPV) has been recognized as an important pathogen for acute respiratory infections in children worldwide and classified into genotypes A and B. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay is a rapid diagnostic method for detecting nucleic acids with a single step under isothermal conditions in less than 1h. RT-LAMP targeting the M gene of hMPV was developed for detecting and identifying hMPV genotypes A and B. The detection limit of the genotype-specific hMPV RT-LAMP assay was 10 times greater than that of conventional reverse transcription polymerase chain reaction (RT-PCR). No cross-reactivity was found with respiratory syncytial virus, parainfluenza virus 1-3, adenovirus, human bocavirus, human rhinovirus, influenza virus A and B, human coronaviruses and enteroviruses. One hundred and fifteen clinical specimens were detected for hMPV genotypes A and B with RT-LAMP, RT-PCR and real-time SYBR PCR. Kappa coefficients showed that there was a good agreement among these three methods. Compared with RT-PCR and real-time SYBR PCR, the genotype-specific RT-LAMP showed better specificity, sensitivity and is more convenient to perform with reduced turn-around time.
authors.affiliation: Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China. Electronic address: songtsinwei@163.com.
authors.forename: Qinwei
authors.initials: Q
authors.lastname: Song
authors.affiliation: Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China. Electronic address: runanzhu@163.com.
authors.forename: Runan
authors.initials: R
authors.lastname: Zhu
authors.affiliation: Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China. Electronic address: sunyu780312@163.com.
authors.forename: Yu
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China. Electronic address: linqingzhao@gmail.com.
authors.forename: Linqing
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China. Electronic address: fangwang1128@sina.com.
authors.forename: Fang
authors.initials: F
authors.lastname: Wang
authors.affiliation: Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China. Electronic address: jie_deng@sina.com.
authors.forename: Jie
authors.initials: J
authors.lastname: Deng
authors.affiliation: Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China. Electronic address: yqianbjc@263.net.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Qian
mesh_terms: D002675:Child, Preschool; D005838:Genotype; D006801:Humans; D007223:Infant; D029121:Metapneumovirus / Q000235:genetics / Q000302:isolation & purification*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D018184:Paramyxoviridae Infections / Q000175:diagnosis* / Q000821:virology; D012141:Respiratory Tract Infections / Q000175:diagnosis / Q000821:virology; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D013696:Temperature; D014773:Virology / Q000379:methods*
keywords: Clinical specimen; Human metapneumovirus; Reverse transcription loop-mediated isothermal amplification