title: Evaluation of real-time RT-PCR compared with conventional RT-PCR for detecting human metapneumovirus RNA from clinical specimens.
abstract: Human metapneumovirus (hMPV) is an etiologic agent of respiratory tract infections. In this study, we compared the sensitivity and specificity of real-time reverse transcription (RT)-polymerase chain reaction (PCR), conventional RT-PCR, and nested PCR in detecting hMPV genes. A total of 146 clinical specimens from 143 patients who showed acute respiratory tract infection symptoms were tested by real-time RT-PCR, conventional RT-PCR, and nested PCR targeting for the fusion gene. We detected hMPV RNA from 14 (9.6%) clinical specimens (real-time RT-PCR, 8; conventional RT-PCR, 5; and nested PCR, 13). When conventional RT-PCR was the reference standard, the sensitivity and specificity of real-time RT-PCR were 100 and 97.9%, respectively. When nested PCR was the standard, the sensitivity and specificity of real-time RT-PCR were 53.8 and 99.2%, respectively. Therefore, real-time RT-PCR was more sensitive than conventional RT-PCR but less so than nested PCR. Phylogenetic analysis showed that the real-time RT-PCR detected four genetic sublineages of hMPV. These results taken together indicate that real-time RT-PCR is an efficient method for detecting four genetic sublineages of hMPV from clinical specimens.
authors.affiliation: Department of Microbiology, Osaka City Institute of Public Health and Environmental Sciences, Osaka, Japan. a-kaida@city.osaka.lg.jp
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kaida
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Kubo
authors.forename: Masashi
authors.initials: M
authors.lastname: Shiomi
authors.forename: Urara
authors.initials: U
authors.lastname: Kohdera
authors.forename: Nobuhiro
authors.initials: N
authors.lastname: Iritani
mesh_terms: D000293:Adolescent; D002648:Child; D002675:Child, Preschool; D006801:Humans; D007223:Infant; D029121:Metapneumovirus / Q000235:genetics / Q000302:isolation & purification*; D008875:Middle Aged; D018184:Paramyxoviridae Infections / Q000821:virology; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D012367:RNA, Viral