Detection of respiratory syncytial virus genome by subgroups-A, B specific reverse transcription loop-mediated isothermal amplification (RT-LAMP)

Masanobu Ushio, Ikuko Yui, Naoko Yoshida, Motoko Fujino, Toshihiro Yonekawa, Yoshinori Ota, Tsugunori Notomi, Tetsuo Nakayama

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

Annual seasonal outbreaks of respiratory syncytial virus (RSV) infection occur every winter. Most patients are diagnosed clinically by a rapid detection kit for RSV protein(s) from nasopharyngeal secretion (NPS), but some problems have been reported on the specificity and sensitivity of such rapid detection kits. To ratify these issues, a sensitive, specific, simple, and rapid molecular based diagnostic method is expected to be introduced and we have developed a method to detect the RSV genome of subgroups A and B independently by reverse transcription loop-mediated isothermal amplification (RT-LAMP). We detected the genomic RNA corresponding approximately to 0.1 TCID 50 in the sample by RT-LAMP for both RSV subgroups under isothermal condition within 60 min after extraction of RNA. Specific DNA amplification was monitored by a real-time turbidimeter and the quantity of RNA was calculated. The RSV genome was detected in 47 of 50 NPS by RT-LAMP, and in 42 by nested RT-PCR, whereas virus isolation was positive for 29 and enzyme-linked immunoassay (EIA) for 34. RSV subgroup A was detected in 25 by RSV RT-LAMP A, RSV subgroup B in 23 by RSV RT-LAMP B, and dual infection with RSV subgroups A and B was identified in one case. They were confirmed with digestion with a specific restriction enzyme, BgI II. The results showed the potential clinical feasibility of RT-LAMP as a useful diagnostic tool for the detection of RSV with high sensitivity similar to nested RT-PCR.

Original languageEnglish
Pages (from-to)121-127
Number of pages7
JournalJournal of Medical Virology
Volume77
Issue number1
DOIs
Publication statusPublished - 2005 Sep
Externally publishedYes

Fingerprint

Respiratory Syncytial Viruses
Reverse Transcription
Genome
Respiratory Syncytial Virus Infections
RNA
Polymerase Chain Reaction
Molecular Pathology
Immunoenzyme Techniques
Disease Outbreaks
Digestion
Viruses
Sensitivity and Specificity
DNA

Keywords

  • Rapid diagnosis
  • Respiratory syncytial virus (RSV)
  • Reverse transcription loop-mediated isothermal amplification (RT-LAMP)
  • Reverse transcription polymerase chain reaction (RT-PCR)
  • Subgroups A and B

ASJC Scopus subject areas

  • Virology

Cite this

Detection of respiratory syncytial virus genome by subgroups-A, B specific reverse transcription loop-mediated isothermal amplification (RT-LAMP). / Ushio, Masanobu; Yui, Ikuko; Yoshida, Naoko; Fujino, Motoko; Yonekawa, Toshihiro; Ota, Yoshinori; Notomi, Tsugunori; Nakayama, Tetsuo.

In: Journal of Medical Virology, Vol. 77, No. 1, 09.2005, p. 121-127.

Research output: Contribution to journalArticle

Ushio, Masanobu ; Yui, Ikuko ; Yoshida, Naoko ; Fujino, Motoko ; Yonekawa, Toshihiro ; Ota, Yoshinori ; Notomi, Tsugunori ; Nakayama, Tetsuo. / Detection of respiratory syncytial virus genome by subgroups-A, B specific reverse transcription loop-mediated isothermal amplification (RT-LAMP). In: Journal of Medical Virology. 2005 ; Vol. 77, No. 1. pp. 121-127.
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