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The development of a diagnostic gene chip for detecting five kinds of viruses in horses

Published online by Cambridge University Press:  20 March 2007

Zhu Lai-hua ,
Liang Cheng-zhu ,
Lu Cheng-ping ,
Wu Hua ,
Yang Yuan-jie ,
Ma Feng-zhong ,
Wang Shu-feng  and
Yu Hong-guang
Zhu Lai-hua
Affiliation:
Nanjing Agricultural University, Nanjing 210095, China Shandong Entry-exit Inspection and Quarantine Bureau, Qingdao 266002, China
Liang Cheng-zhu
Affiliation:
Nanjing Agricultural University, Nanjing 210095, China Shandong Entry-exit Inspection and Quarantine Bureau, Qingdao 266002, China
Lu Cheng-ping*
Affiliation:
Nanjing Agricultural University, Nanjing 210095, China
Wu Hua
Affiliation:
Shandong Entry-exit Inspection and Quarantine Bureau, Qingdao 266002, China
Yang Yuan-jie
Affiliation:
Shandong Entry-exit Inspection and Quarantine Bureau, Qingdao 266002, China
Ma Feng-zhong
Affiliation:
Shandong Entry-exit Inspection and Quarantine Bureau, Qingdao 266002, China
Wang Shu-feng
Affiliation:
Shandong Entry-exit Inspection and Quarantine Bureau, Qingdao 266002, China
Yu Hong-guang
Affiliation:
Shandong Entry-exit Inspection and Quarantine Bureau, Qingdao 266002, China
*
*Corresponding author. E-mail: [email protected] or [email protected]
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Abstract

The highly conserved DNAs of Equine herpesvirus-1 (EHV-1), Equine arteritis virus (EAV), Equine influenza virus (EIV), Equine infectious anaemia virus (EIAV) and Eastern equine encephalomyelitis virus (EEEV) were acquired by molecular cloning, and spotted on the diagnostic gene chip. The cDNAs reverse-transcribed from RNAs of samples were labelled with Cy5-dUTP/Cy3-dUTP as fluorescent probes. Following specific hybridization of the deposited gene chip and labelled probes, fluorescence signals were scanned by laser scanner and the resulting image was analysed by QiamtArray software on a digital computer. The results showed that the prepared gene chip could detect and distinguish the five equine viruses. Its sensitivity was about 25 viral genome copies. The hybridization specificity was confirmed by the presence of red fluorescence signals on the corresponding sites with samples from the five relevant viruses in horses and by the absence of positive signals with the specimens from irrelevant viruses from other animals. Peripheral blood leucocytes (PBL) from some seropositive horses in post-arrival quarantine were negative according to virus isolation, but were positive for EHV-1 and EAV according to the gene chip technique. The overall results suggest that gene chips, which are quick, specific, sensitive and reliable, can provide a practical alternative for screening quarantined animals, and will be able to deal with a large number of animal samples within a very short period of time.

Keywords

Equine herpesvirus-1Equine arteritis virusEquine influenza virusEquine infectious anaemia virusEastern equine encephalomyelitis virusgene chiphybridization
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 3 , Issue 3 , December 2006 , pp. 209 - 215
Copyright
China Agricultural University and Cambridge University Press 2006

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