Diagnosis of human respiratory syncytial virus infection using reverse transcription loop-mediated isothermal amplification.
Human respiratory syncytial virus (RSV) is a major causative agent of lower respiratory tract infections in children and the elderly. A reverse transcription-loop-mediated isothermal amplification (RT-LAMP) was developed assay to amplify the genome of RSV subgroups A and B, in order to improve current diagnostic methods for RSV infection. The primer sets for RT-LAMP were designed using highly conserved nucleotide sequences in the matrix protein region of subgroups A and B, and were specific for each subgroup. The RT-LAMP efficiency was compared to virus isolation and a commercially available enzyme immunoassay (EIA) for RSV detection (BD Directigen EZ RSV test), using nasopharyngeal aspirates from 59 children with respiratory tract infections. The RT-LAMP was specific for RSV and could not detect other respiratory pathogens. 61% (36/59) of children were positive by RT-LAMP, 34% (20/59) by viral isolation, and 56% (26/46) by EZ RSV. Of 16 specimens that were negative by both antigen detection and virus isolation, 12.5% (2/16) were RT-LAMP positive. These results suggest that the RT-LAMP is more sensitive than other methods used to detect RSV. The RT-LAMP assay developed in this study may be useful for diagnostic and epidemiological studies of RSV infection.
Shirato
Kazuya
K
Laboratory of Acute Respiratory Viral Diseases and Cytokines, Department of Virology III, National Institute of Infectious Diseases, Murayama Branch, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan. shirato@nih.go.jp
Nishimura
Hidekazu
H
Saijo
Masayuki
M
Okamoto
Michiko
M
Noda
Masahiro
M
Tashiro
Masato
M
Taguchi
Fumihiro
F
D001483:Base Sequence; D002648:Child; D017931:DNA Primers; D006801:Humans; D008969:Molecular Sequence Data; D009305:Nasopharynx / Q000821:virology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis*; D012136:Respiratory Syncytial Viruses / Q000235:genetics / Q000302:isolation & purification; D048348:Reverse Transcription; D012680:Sensitivity and Specificity
D017931:DNA Primers