鸽圆环病毒全基因组测序及遗传进化分析

doi:10.16431/j.cnki.1671-7236.2025.11.036

摘要/Abstract

摘要： 【目的】了解华东部分地区流行的鸽圆环病毒(Pigeon circovirus，PiCV)基因组特征及遗传变异规律。【方法】无菌采集58只病死鸽的肺脏、脾脏、肝脏组织，利用PCR扩增进行病原检测。对PiCV阳性样本通过PCR扩增病毒基因组全长并进行TA克隆。阳性质粒测序结果通过DNAStar和Mega 11.0软件进行全基因组序列拼接以及核苷酸、氨基酸序列比对，并构建系统进化树。利用RDP 5.42和Simplot 3.5.2软件对分离株进行重组分析并鉴定。【结果】PCR检测结果显示，58只病死鸽中检出30只PiCV阳性，且其他病毒未检出。测序结果显示，成功获得5株PiCV全基因组序列，分别命名为SD02、SD040、JS091、AH056和AH529，序列长度分别为2 037、2 034、2 039、2 039和2 041 bp。相似性比对结果显示，5株PiCV分离株与国内外参考株核苷酸序列相似性为83.2%～100%，与国内的分离株亲缘关系更近。遗传进化分析结果显示，5株PiCV分离株属于不同亚型，分离株SD040、SD02、JZ091、AH056、AH529分别属于B、A、C、G和H亚型。氨基酸序列分析表明，5株PiCV分离株与标准株jz1相比，Cap蛋白氨基酸出现不同程度的突变、替换和缺失，Rep蛋白存在3个氨基酸位点缺失和1个氨基酸位点突变。重组分析表明，5株PiCV分离株有2个重组事件被鉴定，其中分离株AH056是由GX6株和JS2株重组产生，重组断点位于161 bp处。【结论】本研究分离获得5株不同亚型PiCV全基因组序列。病毒重组及关键蛋白突变为其多样性和复杂性的重要成因。本研究丰富了对PiCV基因组特征及遗传演化规律的认识，为华东地区PiCV的防控提供了重要参考依据。

关键词: 鸽圆环病毒; 全基因组测序; 遗传进化分析

Abstract: 【Objective】 The aim of this experiment was to understand the genomic characteristics and genetic variation patterns of Pigeon circovirus (PiCV),which was prevalent in some areas of East China. 【Method】 Sterile samples of lung,spleen,and liver tissues were collected from 58 deceased pigeons and subjected to pathogen detection via PCR amplification.PiCV-positive samples were further subjected to full-length viral genome amplification by PCR and TA cloning.The positive plasmid sequencing results were used for full-genome sequence assembly,nucleotide and amino acid sequence alignment through DNAStar and Mega 11.0 software,and the phylogenetic tree was constructed.Recombination analysis and identification of the isolates were performed using RDP 5.42 and Simplot 3.5.2 software. 【Result】 PCR test results showed that among the 58 dead pigeons,30 were positive for PiCV,and no other viruses were detected.The sequencing results showed that the whole genome sequences of five PiCV isolates were successfully obtained,named SD02,SD040,JS091,AH056, and AH529 respectively,and the sequence lengths were 2 037,2 034,2 039,2 039, and 2 041 bp, respectively.The similarity comparison results showed that the nucleotide sequence similarity of the five PiCV isolates to the reference sequences registered both domestically and internationally ranged from 83.2% to 100%,and they were more closely related to the domestic isolates.Genetic evolution analysis showed that the five PiCV isolates belonged to different subtypes,with isolates SD040,SD02,JZ091,AH056,and AH529 belonged to subtypes B,A,C,G, and H,respectively.Amino acid sequence analysis indicated that compared with the standard strain jz1,the Cap protein amino acids of the five PiCV isolates had different degrees of mutations,substitutions and deletions,and the Rep protein had three amino acid deletions and one amino acid mutation.Recombination analysis identified two recombination events among the five PiCV isolates,with isolate AH056 being a recombinant of GX6 and JS2 strains,with a recombination breakpoint located at 161 bp. 【Conclusion】 In this study,the complete genomic sequences of five different subtypes of PiCV were isolated and obtained.Viral recombination and mutations in key proteins were significant contributors to their diversity and complexity.This study enriched the understanding of the genomic characteristics and genetic evolution patterns of PiCV and provided important reference data for the epidemiological investigation of PiCV in East China.

Key words: Pigeon circovirus; whole genome sequencing; genetic evolution analysis

中图分类号:

S855.3

王仲浩, 田野, 袁小远, 李桂明, 陈书民, 刘明超, 王传文, 孟凯. 鸽圆环病毒全基因组测序及遗传进化分析[J]. 中国畜牧兽医, 2025, 52(11): 5414-5421.

WANG Zhonghao, TIAN Ye, YUAN Xiaoyuan, LI Guiming, CHEN Shumin, LIU Mingchao, WANG Chuanwen, MENG Kai. Whole Genome Sequenccing and Genetic Evolution Analysis of Pigeon Circovirus[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(11): 5414-5421.

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