Skip to main content Accessibility help
×
Home

Establishment of multiple RT-PCR diagnostic techniques for Avian influenza virus (AIV), Newcastle disease virus (NDV), Classical swine fever virus (CSFV) and Foot-and-mouth disease virus (FMDV)

  • Liu Wei-Quan (a1), Liu Hai-Peng (a2), Jiang Yu (a3), Wang Ben-Xu (a3), Wang Ji-Gui (a1), Yang Shu-Yan (a1), Sun Shao-Guang (a1), Tu Chang-Chun (a3) and Liu Peng-Peng (a1)...

Abstract

The homology of the sequences, reported and registered in GenBank, of different strains of Avian influenza virus (AIV), Newcastle disease virus (NDV), Classical swine fever virus (CSFV) and Foot-and-mouth disease virus (FMDV), was analysed and compared with each other. According to the properties of these viruses, the conservative domain of the M gene for AIV, the F gene for NDV, the 5' non-coding domain end for CSFV and the 2B gene for FMDV were selected for polymerase chain reaction (PCR) amplification. In order to prevent the formation of conformational dimers between different primers, four pairs of primers designed with the DNAsis system under the condition of G+C (50–60%),18–25 bp in length and Tm (72–85), were analysed using the VNTI5.5 system. The specific fragments amplified were as follows: 141 bp for FMDV, 200 bp for CSFV, 319 bp for NDV and 471 bp for AIV. The optimal conditions of PCR for each virus mentioned above were determined by orthogonal assay, and two or four of the four pairs of primers were then combined and used for amplification trials. The results showed that four specific fragments of different lengths would be successfully amplified in one tube at the same time. The products of PCR were tested to be specific by sequencing. Out of 46 pathological samples detected with the multiple PCR, there were 5 AIVs, 7 NDVs, 15 CSFVs and 6 FMDVs. The amplification above was identified with a single PCR. On the other hand, the results corresponded to those of electronic microscopy, haemagglutination (HA) and enzyme-linked immunosorbent assay (ELISA). The method described here is practicable, sensitive, specific, simple and cheap. It could be used for diagnosing AIV, NDV, CSFV and FMDV in different animals.

Copyright

Corresponding author

*Corresponding author: Email weiquan1@yahoo.com

References

Hide All
Harding, M, Lutze-Wallace, C, Prud'Homme, I et al. , (1994) Reverse transcriptase-PCR assay for detection of hog cholera virus. Journal of Clinical Microbiology 32: 26002602.
International Organization of Animal Health (1999) The International Code of Animal Health. Beijing: Bingqi Industry Press (translated by The Bureau of Animal Husbandry and Veterinary Sciences in China, Agricultural Ministry, 2000).
Liu, HP, Liu, WQ, Jiang, Y et al. , (2003a) The establishment of multiple RT-PCR diagnostic techniques for CSFV and FMDV. China Animal Husbandry and Veterinary Medicine 3(2): 3336.
Liu, HP, Liu, WQ, Jiang, Y et al. , (2003b) The establishment of multiple RT-PCR diagnostic techniques for AIV and NDV. Chinese Journal of Veterinary Science and Technology 33(7): 942.
Meyer, RF, Brown, CC, House, C et al. , (1991) Rapid and sensitive detection of foot-and-mouth disease virus in tissues by enzymatic RNA amplification of the polymerase gene. Journal of Virological Methods 34: 161172.
Naffakh, N, Massin, P, Escriou, N et al. , (2000) Genetic analysis of the compatibility between polymerase proteins from human and avian strains of influenza A viruses. Journal of General Virology 81: 12831291.
Seal, BS, King, DJ and Bennett, JD (1995) Characterization of newcastle disease virus isolates by reverse transcription PCR coupled to direct nucleotide sequencing and development of sequence database for pathotype prediction and molecular epidemiological analysis. Journal of Clinical Microbiology 33: 26242630.
Vandenvelde, C, Verstraete, M and Van Beers, D (1990) Fast multiplex polymerase chain reaction on boiled clinical samples for rapid viral diagnosis. Journal of Virological Methods 30(2): 215227.
Yamada, A, Imanishi, J, Nakajima, E et al. , (1991) Detection of influenza viruses in throat swabs by using polymerase chain reaction. Microbiology and Immunology 35(3): 259265.
Yin, Z and Liu, JH (1997) Animal Virology, 2nd ed. Beijing: Science Press.

Keywords

Related content

Powered by UNSILO

Establishment of multiple RT-PCR diagnostic techniques for Avian influenza virus (AIV), Newcastle disease virus (NDV), Classical swine fever virus (CSFV) and Foot-and-mouth disease virus (FMDV)

  • Liu Wei-Quan (a1), Liu Hai-Peng (a2), Jiang Yu (a3), Wang Ben-Xu (a3), Wang Ji-Gui (a1), Yang Shu-Yan (a1), Sun Shao-Guang (a1), Tu Chang-Chun (a3) and Liu Peng-Peng (a1)...

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed.