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 journalArticlepeer-review

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

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
  • Infectious Diseases

Fingerprint Dive into the research topics of 'Detection of respiratory syncytial virus genome by subgroups-A, B specific reverse transcription loop-mediated isothermal amplification (RT-LAMP)'. Together they form a unique fingerprint.

Cite this