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.
- Author:
Sang Wook PARK
1
;
Tae Won KWON
;
Eun Soon KIM
;
Young Dae WOO
;
Yoon Suk KIM
;
Yoo Kyum KIM
Author Information
1. Department of Virology, Asan Institute for Life Sciences, Korea.
- Publication Type:Original Article
- Keywords:
Respiratory syncytial virus;
Parainfluenza virus;
Multiplex RT-PCR;
Indirect immunofluorescence;
Shell vial assay
- MeSH:
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*
- From:Journal of Bacteriology and Virology
2002;32(2):203-210
- CountryRepublic of Korea
- Language:Korean
-
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.
