信阳地区散养土鸡禽白血病病毒和马立克病病毒混合感染的分子生物学诊断

doi:10.16431/j.cnki.1671-7236.2025.02.034

Abstract

Abstract: 【Objective】 The objective of this experiment was to investigate the co-infection and genetic evolution of Avian leukosis virus (ALV) and Marek’s disease virus (MDV) in free range local chickens in Xinyang area. 【Method】 Pathological autopsy was performed on the chickens suspected to be co-infected with ALV and MDV,and PCR amplification was performed using specific primers.The ALV-J subgroup (ALV-J) gp85 gene and MDV meq gene were amplified and sequenced,and the nucleotide and amino acid sequences similarity and amino acid variation sites were analyzed by MegAlign software.The phylogenetic tree of gp85 and meq amino acid sequences was constructed with Mega 11.0 software. 【Result】 The chickens were examined for enlargement of the gland,stomach and spleen,and diffuse enlargement of the liver with grayish white nodules.PCR results of ALV-J gp85 and MDV 132-bpr genes were positive, and 545 and 317 bp target bands were obtained, respectively. Sequencing results showed that the amplified sequences were highly similar to the gp85 gene of ALV-J strain GX12NN04 (accession number: KT598488) and the meq gene of MDV-1 strain YLO40920 (accession number: DQ174459), and the similarity was 94.6% and 99.9%, respectively. The isolates were initially identified as ALV-J and MDV,and named as HN23XY01-ALV and HN23XY01-MDV,respectively.Nucleotide sequence analysis showed that the nucleotide sequence similarity between HN23XY01-ALV gp85 gene and 13 ALV-J reference strains was 92.1%-94.6%，and the nucleotide sequence similarity between HN23XY01-MDV meq gene and 14 MDV reference strains was 99.3%-99.9%.The amino acid sequence analysis showed that the amino acid sequence similarity between HN23XY01-ALV gp85 and 13 ALV-J reference strains was 87.4%-91.4%,and the amino acid sequence similarity between HN23XY01-MDV and 14 MDV reference strains was 97.9%-99.7%.Phylogenetic tree results showed that HN23XY01-ALV and ALV-J were closely related and in the same evolutionary branch.HN23XY01-MDV was in the same branch with Guangxi isolates GXY2,YLO40920 and GX20NN2，and Henan isolate HNSC105,and was distantly related to vaccine strains CVI988,814 and CU-2.Analysis of amino acid variation sites showed that HN23XY01-ALV had mutations at D(E)65Y,Q(K/R)75L,A(T)76S,R(T)119K and T(M/A)219K.HN23XY01-MDV had K77E,D80Y,T139A,P176R and P217A amino acid mutations，and contained site 193 (proline) that was missing from the vaccine strains,which was consistent with the characteristics of MDV-1 virulent strains. 【Conclusion】 In this study,co-infection of ALV-J and MDV was detected in free range local chickens in Xinyang area，and the results provided important references for the study of the co-pathogenic mechanism of ALV-J and MDV in local chickens and the epidemic prevention and control of co-infection.

Key words: Avian leukosis virus (ALV); Marek’s disease virus (MDV); gp85 gene; meq gene; co-infection

CLC Number:

S855.3

LI Yingxiao, YIN Lei, ZHAO Yu, DONG Jianguo, HE Shuhai, QU Zhehui, JIAO Fengchao. Molecular Biological Diagnosis of Co-infection of ALV and MDV in Free Range Local Chickens in Xinyang Area[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(2): 844-854.

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Molecular Biological Diagnosis of Co-infection of ALV and MDV in Free Range Local Chickens in Xinyang Area

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