宁夏地区绵羊博尔纳病病毒自然感染状况研究

BDV Natural Infection in Sheep in Ningxia

目的探讨中国宁夏地区绵羊博尔纳病病毒(Borna disease virus,BDV)的自然感染状况,并分析比较中国宁夏地区绵羊自然感染BDVp24基因序列与国外人和动物来源BDV分离株及其标准株StrainV和He/80的同源性。方法采用巢式逆转录荧光定量PCR(FQ-nRT-PCR技术)检测了中国宁夏地区268例绵羊PBMCs中BDVp24基因片段,对阳性标本FQ-PCR产物进行基因序列测定,测序结果应用BLAST软件以及DNAsist5.0软件对测序结果分析,与国外人与动物来源的BDV分离株以及标准株StrainV和He/80进行序列比较。结果268例中4例绵羊外周血BDVp24基因片段阳性,阳性率为1.49%;测序分析结果表明,宁夏地区绵羊感染的BDVp24的核苷酸序列与马源性病毒株H3575同源性最近,同源性为98.84%(85/86),与标准株StrainV和He/80同源性为94.19%和100%。并且它们编码的氨基酸序列相同。结论宁夏地区绵羊中可能存在动物源性的BDV自然感染,该地区感染的BDVp24核苷酸序列与马源性病毒株H3575、标准株StrainV和He/80存在高度的同源性,其感染来源可能相同亦或存在相互之间感染的可能。

Objective To investigate the natural infection of BDV in sheep in Ningxia and to explore if the nucleotide sequence in BDVp24 was homophyhc with the overseas strains, including standard strain V/FR and He/80 detected from human and animals. Methods The p24 fragment of BDV was detected by nested reverse transcriptase polymerase chain reaction with real-time PCR（Real-time nRT-PCR） in PBMCs from 268 sheep. The products of Real-time nRT-PCR from positive samples were cloned and sequenced before compared with those of the overseas strains, including strain V/FR and strain He/80 by BLAST and DNAsist 5.0. Results There were 4 positive samples in 268 sheep. The gene sequence for positive product showed BDV p24 in sheep from Ningxia was homophylic with strain V/FR（94.19% ）and strain He/80 detected from ill horses and strain H3575（98.84% ）. However, their amino acid sequence remained the same. Conclusion There is BDV infection, which probably originate from animals including sheep in Ningxia. The gene sequence is homophylic with that of standard strain V/FR, strain He/80 and strain H3575. The Source of infection may be the same, or the existence of possible infection with Each other.