J Bacteriol Virol.  2002 Jun;32(2):203-210.

Rapid Detection and Identification of Human Respiratory Syncytial Virus, Human Parainfluenza Virus Type 1, 2 and 3 by Single-tube Multiplex Reverse Transcription Polymerase Chain Reaction

Affiliations
  • 1Department of Virology, Asan Institute for Life Sciences, Korea.
  • 2Department of Vascular Surgery, University of Ulsan College of Medicine, Poongnap-dong, Songpa-gu, Seoul, Korea. ykkim@amc.seoul.kr
  • 3Department of Biochemistry, University of Ulsan College of Medicine, Poongnap-dong, Songpa-gu, Seoul, Korea.
  • 4Department of Microbiology, University of Ulsan College of Medicine, Korea.

Abstract

Laboratory diagnosis of respiratory viral infection has traditionally been based upon virus isolation and/or viral antigen identification. Recently, more sensitive and specific nucleic acid detection methods by reverse transcription- polymerase chain reaction (RT-PCR) have been developed, however, conventional RT-PCR can identify only a single suspected virus. To identify the causative agents which belong to Paramyxoviridae of respiratory virus infections, we have developed a single-tube multiplex RT-PCR using four primer sets which can amplify respiratory syncytial virus and parainfluenza virus type 1, 2 and 3 simultaneously. Assay sensitivity of single-tube multiplex RT-PCR allowed a detection in the range of 3~500 TCID50 and there were no cross amplification among other respiratory viral agents based on the test using reference virus stocks. The single-tube multiplex RT-PCR was able to directly detect viruses in respiratory specimens, with virus being detected 11 of 80 samples as compared to 9 of 80 samples detected by indirect immunofluorescence or antigen detection following shell vial culture. This result suggests that the single-tube multiplex RT-PCR can be established as a more sensitive and rapid diagnostic application than shell vial assay for the detection of respiratory infection of Paramyxoviridae.

Keyword

Respiratory syncytial virus; Parainfluenza virus; Multiplex RT-PCR; Indirect immunofluorescence; Shell vial assay

MeSH Terms

Clinical Laboratory Techniques
Fluorescent Antibody Technique, Indirect
Humans*
Parainfluenza Virus 1, Human*
Paramyxoviridae
Paramyxoviridae Infections*
Polymerase Chain Reaction*
Respiratory Syncytial Virus, Human*
Respiratory Syncytial Viruses
Reverse Transcription*
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