Hostname: page-component-78c5997874-lj6df Total loading time: 0 Render date: 2024-11-15T09:22:05.205Z Has data issue: false hasContentIssue false
  • English
  • Français

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)

Published online by Cambridge University Press:  12 February 2007

Liu Wei-Quan ,
Liu Hai-Peng ,
Jiang Yu ,
Wang Ben-Xu ,
Wang Ji-Gui ,
Yang Shu-Yan ,
Sun Shao-Guang ,
Tu Chang-Chun  and
Liu Peng-Peng
Liu Wei-Quan*
Affiliation:
College of Biology, China Agricultural University, Beijing 100094, China
Liu Hai-Peng
Affiliation:
Shenyang Agricultural University, Shenyang 110161, China
Jiang Yu
Affiliation:
The Military Veterinary Institute of PLA, Changchun 130062, China
Wang Ben-Xu
Affiliation:
The Military Veterinary Institute of PLA, Changchun 130062, China
Wang Ji-Gui
Affiliation:
College of Biology, China Agricultural University, Beijing 100094, China
Yang Shu-Yan
Affiliation:
College of Biology, China Agricultural University, Beijing 100094, China
Sun Shao-Guang
Affiliation:
College of Biology, China Agricultural University, Beijing 100094, China
Tu Chang-Chun
Affiliation:
The Military Veterinary Institute of PLA, Changchun 130062, China
Liu Peng-Peng
Affiliation:
College of Biology, China Agricultural University, Beijing 100094, China
*
*Corresponding author: Email [email protected]
Get access Rights & Permissions [Opens in a new window]

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.

Keywords

Avian influenza virusNewcastle disease virusClassical swine fever virusFoot-and-mouth disease virusmultiple RT-PCR
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 2 , Issue 3 , December 2005 , pp. 161 - 165
Copyright
Copyright © China Agricultural University and Cambridge University Press 2005

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

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: 2600–2602.CrossRefGoogle ScholarPubMed
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).Google Scholar
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): 33–36.Google Scholar
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): 9–42.Google Scholar
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: 161–172.CrossRefGoogle ScholarPubMed
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: 1283–1291.Google ScholarPubMed
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: 2624–2630.CrossRefGoogle ScholarPubMed
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): 215–227.CrossRefGoogle ScholarPubMed
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): 259–265.CrossRefGoogle Scholar
Yin, Z and Liu, JH (1997) Animal Virology, 2nd ed. Beijing: Science Press.Google Scholar