关键词: 规模化鹅场, 核酸检测, 遗传进化分析

Abstract:

【Objective】The author’s research group has long investigated the nucleic acid detection rate of common viral diseases in scaled goose farms in various provinces and cities in China, and found that many large-scaled goose farms in China could detect multiple viral nucleic acids at the same time, and this phenomenon was very common. In view of the lack of relevant data on the simultaneous detection of multiple viral diseases in large-scale goose farms in China, the purpose of this paper was to understand and analyze the simultaneous detection of nucleic acids of multiple viral diseases in large-scaled goose farms, so as to provide the theoretical guidance and scientific basis for the prevention and control of viral diseases in scaled goose farms.【Method】From May 2021 to October 2021, 737 diseased materials were collected from 47 scaled goose farms in Shandong, Heilongjiang, Sichuan, Jilin, Guangxi, Henan, Anhui, Liaoning, Hebei, Guizhou, Hunan and Inner Mongolia. These samples were detected for Muscovy duck reovirus (MDRV), duck reovirus (DRV), goose astroviruses (GAstV), fowl adenovirus (FAdV), reticuloendotheliosis virus (REV), Newcastle disease (NDV), goose parvovirus (GPV), goose circovirus (GoCV), goose hemorrhagic polyomavirus (GHPV), Tembusu virus (TMUV) and H9 subtype avian influenza virus (H9-AIV) by ordinary PCR and RT-PCR. The liver, spleen, lung and kidney were dissected from each sample, and total RNAs were extracted by Trizol method; a strand of cDNA was synthesized by reverse transcription with total RNA as the template, and then the complete cDNA was obtained by continuous amplification with cDNA as the template. Using these cDNA as template, the specific primers of MDRV, DRV, GAstV, FAdV, REV, NDV, GPV, GoCV, GHPV, TMUV and H9-AIV were used to amplify the target fragment by ordinary PCR reaction. All amplified fragments were analyzed by agarose gel electrophoresis, and some positive samples were sequenced. The obtained sequencing results were compared with the corresponding virus gene sequences published on GenBank, and the phylogenetic tree was drawn by Neighbor-Joining (N-J) method in MEGA 6.0 software for analysis.【Result】MDRV, DRV, GAstV, FAdV, REV, NDV, GPV, GoCV, GHPV, TMUV and H9-AIV showed that the detection rate of GAstV was the highest, with the value of 58.21%; the detection rates of REV and NDV were the lowest, with the value of 1.36% and 1.50%, respectively. A variety of viral nucleic acids could be detected simultaneously in geese to varying degrees, especially in two or three viral nucleic acids, accounting for 67.44% of the total samples. In the simultaneous detection rate of nucleic acids of the two viruses, GAstV and GoCV accounted for the largest proportion, which was 18.44%; MDRV, GAstV and GPV accounted for 36.28% of the simultaneous detection rates of the three virus nucleic acids.【Conclusion】It was further confirmed that multiple viral nucleic acids could be detected simultaneously in large-scaled goose farms in China. It was speculated that this might be one of the important reasons for the complexity of viral diseases and the difficulty of prevention and control in large-scaled goose farms in China.

Key words: scaled goose farm, nucleic acid test, phylogenetic analysis