病毒宏基因组学检测猪博卡病毒及其遗传进化分析  

Detection of Porcine Bocavirus by Viral Macrogenomics and Analysis of its Genetic Evolution

作　　者：孙晓瑜 单虎[1,2,3] 黄娟 SUN Xiaoyu;SHAN Hu;HUANG Juan(College of Veterinary Medicine,Qingdao Agricultural University,Qingdao 266109,China;Shandong Provincial Key Laboratory of Preventive Veterinary Medicine,Qingdao 266109,China;Shandong Provincial Veterinary Medicine Diagnostic Reagent Engineering Technology Research Center,Qingdao 266109,China)

机构地区：[1]青岛农业大学动物医学院,山东青岛266109 [2]山东省预防兽医学重点实验室,山东青岛266109 [3]山东省兽药诊断试剂工程技术研究中心,山东青岛266109

出　　处：《中国兽医杂志》2024年第4期50-57,共8页Chinese Journal of Veterinary Medicine

基　　金：山东省生猪产业技术体系(SDAIT-08-07)。

摘　　要：为了探明导致某养猪场中仔猪腹泻的病原,本试验将患病仔猪样本处理并提取核酸后进行病毒宏基因组测序,对测序结果进行序列比对分析和遗传进化分析,并对样本核酸进行PCR检测验证。结果显示,RNA样本中只注释到A型流感病毒(H1N1和H3N2),读数比为6.8%,未发现其他猪病相关RNA病毒。DNA样本注释到的病毒主要为猪博卡病毒(PBoV),读数比为34.5%,命名为PBoV-SDPD-2022;其余猪病相关病毒包括猪巨细胞病毒、猪乳腺腺病毒和猪细环病毒,读数比均小于1.0%。基于全基因组、NS 1基因和VP 1基因构建的系统发育进化树均显示,PBoV-SDPD-2022与参考毒株PBoV3_VIRES_HuB01_C1处于同一分支,属于PBoV G3群。与21株参考毒株相比,PBoV-SDPD-2022 NS1蛋白的编码基因核苷酸和氨基酸序列同源性分别为49.6%~97.2%和38.5%~95.3%;VP1蛋白的编码基因核苷酸和氨基酸序列同源性分别为47.9%~97.9%和34.8%~98.8%;NP1蛋白的编码基因核苷酸和氨基酸序列同源性分别为35.2%~98.5%和26.9%~97.4%。与G3群参考毒株相比,PBoV-SDPD-2022 NS1蛋白在第654位点处有1个氨基酸的缺失;VP1蛋白在第151、152位点处有2个氨基酸的插入,且包含基序YLGPF(VPl aa 18~22)和HDLAY(VP1 aa 41~45);NP1蛋白在第15、16、35、36、212位点处发生氨基酸的缺失,在第95位点处有1个氨基酸的插入。PCR检测证实样本核酸为PBoV阳性。结果表明,导致该养猪场仔猪腹泻的优势病原是G3群PBoV。本试验有助于进一步了解我国PBoV基因组序列特征,并为未来PBoV流行病学调查提供了参考依据。In order to explore the pathogen causing diarrhea in piglets on a pig farm,this study processed and extracted nucleic acids from diseased piglet samples for virus metagenomic sequencing.The sequencing results were analyzed for sequence alignment and genetic evolution,and nucleic acid samples were also validated by PCR.The results showed that RNA samples only annotated to A-type influenza viruses(H1N1 and H3N2)with a read ratio of 6.8%,and no other porcine-related RNA viruses were found.The annotated viruses in DNA samples were mainly porcine bocavirus(PBoV)with a read ratio of 34.5%,named as PBoV-SDPD-2022;other porcine-related viruses included porcine cytomegalovirus,porcine mammary adenovirus,and porcine torque teno virus,all with read ratios less than 1.0%.Phylogenetic trees constructed based on the full genome,NS 1 gene,and VP 1 gene all showed that PBoV-SDPD-2022 and the reference strain PBoV3_VIRES_HuB01_C1 were in the same branch,belonging to the PBoV G3 group.Compared with 21 reference strains,the coding gene nucleotide and amino acid sequence homology of PBoV-SDPD-2022 NS1 protein were 49.6%-97.2%and 38.5%-95.3%,respectively.For the VP1 protein,the coding gene nucleotide and amino acid sequence homology were 47.9%-97.9%and 34.8%-98.8%,respectively.For the NP1 protein,the coding gene nucleotide and amino acid sequence homology were 35.2%-98.5%and 26.9%-97.4%,respectively.Compared with the G3 group reference strains, PBoV-SDPD-2022 NS1 proteinhadone amino acid deletion at position 654;VP1 protein had two amino acid insertions at positions 151 and 152, including the motifs YLGPF (VPl aa 18-22) and HDLAY (VP1 aa 41-45);NP1 protein had amino acid deletions at positions 15, 16, 35, 36, and 212, and one amino acid insertion at position 95. PCR detection confirmed that the nucleic acid samples were positive for PBoV. The results indicated that the predominant pathogen causing diarrhea in piglets on the pig farm was the G3 group of PBoV. This study contributes to a better understanding of the genomic characterist

关 键 词：病毒宏基因组学 猪博卡病毒 遗传进化分析 

分 类 号：S85[农业科学—兽医学]