猪圆环病毒2型的分离鉴定及其全基因组序列分析

Isolation and Identification of Porcine Circovirus Type 2 and Its Complete Genome Sequence Analysis

为分离鉴定流行于河南省的猪圆环病毒2型(PCV2),对河南省不同地区猪场的11份疑似断奶仔猪多系统衰竭综合征(PMWS)猪的病料进行分离鉴定,得到PCV2毒株。通过PCR检测,11份临床疑似病料中,7份确诊为PCV2阳性病料,阳性率为63.6%。挑选出阳性值较高的毒株命名为zmd-20161022,通过增殖培养筛选出优良毒株,并对其进行单层过氧化物酶试验(IPMA)鉴定,分析免疫原性;应用PCR对该病毒全基因组进行特异性扩增,经测序后对该病毒的同源性和系统进化进行分析。结果显示,该毒株序列全长1 767 bp,同源性分析发现与国内3株PCV2毒株(AY686763、HM641752、HM038034)的同源性为95.2%~98.3%,与基因登录号为HM038030.1的同源性高达98.6%。系统进化树分析发现,zmd-20161022毒株与基因登录号为HM038030.1的亲缘关系很近,被鉴定为PCV2b-1c型,属于PCV2b的一种亚型。

In order to isolate and identify porcine circovirus type 2,11 clinical samples from weanling pigs suspected of PCV2 infection,which is endemic to pig farms in Henan province,were tested and identified.The 7 out of 11 clinical samples were diagnosed as PCV2 positive by PCR,and the positive rate was 63.6%.The strain with the highest positive value was selected and named as zmd-20161022,which was further tested by IPMA to analyze the immunogenicity,and the complete genome was amplified by PCR and sequenced for homology and phylogenetic analysis.The full length of zmd-20161022 was 1 767 bp,95.2%-98.3%homology with three PCV2 strains(AY686763,HM641752,HM038034)from China,and 98.6%homology with the PCV2 strain(the gene accession number HM038030.1)in GenBank.Phylogenetic analysis showed that the zmd-20161022 strain belonged to PCV2b-1c,a subtype of PCV2b,which was closely related to the gene accession number HM038030.1.