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One-step multiplex RT-PCR for detection and subtyping of swine influenza H1, H3, N1, N2 viruses in clinical samples using a dual priming oligonucleotide (DPO) system
Cited 35 time in
Web of Science
Cited 35 time in Scopus
- Authors
- Issue Date
- 2008-05-19
- Publisher
- Elsevier
- Citation
- J. Virol. Methods 151, 30-34
- Keywords
- Swine influenzavirus ; Multiplex PCR ; DPO
- Abstract
- The swine influenza virus (SIV) H1N1, H1N2, and H3N2 subtypes circulate in Korean farm. A novel multiplex
RT-PCR (m-RT-PCR) was developed to detect and subtype swine influenza viruses. This m-RT-PCR
assay could identify H1, H3, N1 and N2 from clinical samples in single tube reaction using DPO system.
Korean SIVs are closely related to the United States influenza viruses, and primers were developed
for SIV from North American viruses and recently Korean isolates. The sensitivity of the m-RT-PCR was
10TCID50/ml for H1N1, H1N2 or H3N2. The lowest viral concentrations detected by single PCR were
1TCID50/ml for each subtype. Non-specific reactions were not observed when other viruses and bacteria
were used to assess the m-RT-PCR. The results of m-RT-PCR were more effective than virus isolation or
hemagglutination (HA) test. This assay using a DPO system provides a rapid, sensitive, and cost-effective
laboratory diagnosis for detecting and subtyping of SIV in pigs.
- ISSN
- 0166-0934
- Language
- English
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