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机构地区:[1]宁夏医科大学附属石嘴山市第一人民医院,石嘴山753200 [2]宁夏医科大学附属医院神经内科
出 处:《中华流行病学杂志》2009年第12期1284-1287,共4页Chinese Journal of Epidemiology
摘 要:目的 检测宁夏地区不明原因病毒性脑炎患者外周血单个核细胞(PBMCs)博尔纳病病毒(BDV)携带情况,分析BDV与标准病毒株之间的同源性。探讨BDV感染性疾病与不明原因病毒性脑炎之间的关系,同时也为部分病毒性脑炎的病因学诊断及其防治提供实验依据。方法采用荧光定量巢式反转录酶聚合酶链反应(FQ—nRT-PCR)检测宁夏地区不明原因病毒性脑炎患者及正常人PBMCsBDVp24基因片段,并对其中阳性扩增产物进行基因序列测定,然后采用BLAST软件以及DNAsist5.0软件对测序结果进行分析。结果病毒性脑炎组样本BDVp24基因片段FQ.nRT-PCR阳性率(10.17%)显著高于正常对照组(0%)(P〈0.05)。病毒性脑炎患者检出BDVp24基因片段的核苷酸序列与马BDV标准病毒株H3915序列同源性为97.67%,与H1766株比较在3个位点出现一致性突变(nt1659T-c,nt1668A→G,nt1674T→C;突变率为3.49%);与strainV株比较在4个位点出现一致性突变(nt1648C→T,nt1658e→A,nt1667A—G,nt1673T→C;突变率为4.65%),但编码的氨基酸相同。证实扩增所得目的基因片段确系BDVp24的相应片段。结论宁夏地区不明原因病毒性脑炎患者中存在BDV感染,提示部分不明原因病毒性脑炎的发生可能与BDV感染有关。且该BDVp24扩增产物核苷酸序列与标准病毒株高度同源性,与马源strainH3915同源性最高。Objective To investigate the infection of Borna disease virus (BDV) in unidentified viral encephalitis patients in Ningxia in order to explore if the nucleotide sequence and amino acid sequence in BDV p24 were homophylic with the overseas standard strain. We also intended to investigate the correlation between BDV infection and the unidentified viral encephalitis patients from Ningxia to lay an experimental basis for etiological diagnosis, prevention and treatment on certain human neuropsychiatric disorders. Methods BDV p24 gene fragment was detected by nested reverse transcriptase polymerase chain reaction with fluorescence quantitative PCR (FQnRT-PCR) in peripheral blood mononuclear cells (PBMCs). Samples were from 59 unidentified viral encephalitis patients and 60 normal controls. For those positive products, gene sequence and amino acid sequence were then analyzed by BLAST and DNAsist 5.0. Results The positive rate of the BDV p24 gene fragment in PBMCs from the unidentified viral encephalitis (10.17%) was significantly higher than from the controls (0%) (P〈0.05). Data from the gene sequence on those positive products showed BDV p24 fragment in the patients with unidentified viral encephalitis from Ningxia was homophylic with strain H3915 detected from ill horses (97.67%), as well as with the strain H1766 (96.51%) and strain V (95.35%). However, their amino acid sequences remained the same. Conclusion BDV infection might probably have existed in the unidentified viral encephalitis patients of Ningxia. The gene sequence seemed to be homophylic with that of standard strain H1766 and strain V, especially with strain H3915.
关 键 词:博尔纳病病毒 病毒性脑炎 荧光定量巢式反转录酶聚合酶链反应
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