病毒性脑炎患者BDVp24、p40基因片段的核酸检测  

Detection of nucleic acid of borna disease virus p24 and p40 gene fragments in patients with viral encephalitis

出　　处：《中国医师杂志》2013年第12期1605-1608,共4页Journal of Chinese Physician

基　　金：湖南省科技基金资助项目（2010SK3203）

摘　　要：目的分别检测病毒性脑炎患者脑脊液有核细胞（CSFMCs）中博尔纳病病毒（BDV）p24、p40基因片段，探讨病毒性脑炎患者与BDV感染的关系及阳性患者的临床特征。方法用荧光定量巢式逆转录聚合酶链反应（FQ—nRT—PCR）方法检测病毒性脑炎患者及对照者CSFMCs中BDVp24、p40基因片段，将两个保守区p24和p40检测均为阳性的标本再做进一步分析，并总结阳性患者的临床特征，同时用β-肌动蛋白（B—actin）作为内参照。结果实验组102例病毒性脑炎患者CSFMCs中BDVp24基因片段检测中有11例阳性，BDVp40基因片段检测中有10例阳性，两个保守区p24和p40检测均为阳性标本10例，对照组84例中均为阴性，两者比较阳性率差异有统计学意义（P〈0．05）。阳性基因片段产物测序后与BDV标准病毒株V和马源的BDV病毒株H1766序列比较，同源性分别为96．35％和98．85％，在4个位点出现基因突变（nt1649T→C，nt1656G→A，nt1670C→T，nt1676C→T）；8例阳性患者主要以精神行为异常为起病方式，患者与动物关系密切，其他临床特征无特殊。结论病毒性脑炎的发生与BDV感染有关，主要以精神行为异常为临床特征。Objective To detect Borna disease virus p24 and p40 gene fragments from cerebrospinal fluid mononuclear cells （CSFMCs） of the patients with viral encephalitis, and to investigate clinical features of patients infected by borua disease virus （BDV） and their relationship with BDV. Methods The p24 and p40 gene fragments of BDV in CSFs in these patients infected in viral encephalitis were examined by nested reverse transcriptase polymerase chain reaction （RT-PCR） and fluorescence quantitative （FQ） PCR, and control groups were also examined with same method. At the same time, β-actin was detected as internal reference. The clinical characteristics of positive patients infected by BDV were summarized. Resuits The number of the positive sample of BDV p24 was ten and BDV p40 was eleven in experimental group. There are no positive patients of BDV p24 and p40 in control group. The positive rate in experimental group was significantly higher than that in control group （ P 〈 0. 05 ）. The number of the positive patients of both BDV p24 and BDV p40 in CSFMCs was ten in 102 patients. Homology was 95.35% and 98. 84%, which had 4 situs of consistent silent mutation （nt1649 T→C, nt1656 G→A, nt1670 C→T, and nt1676 C→T）, compared with the sequence of the standard strain V and the virus strain H1766 of BDV from horse supplied by GenBank. The clinical features of ten viral encephalitis patients infected BDV were mainly mental and behavior disorders, and were genetically connected with animals, the other clinical findings were the same as VE. Conclusions The results suggest that BDV infection possibly exist in viral en-cephalitis patients, and the clinical features of viral encephalitis patients infected BDV be mainly mental and behavior disorders.

关 键 词：脑炎 病毒性 遗传学 BORNA病病毒 遗传学 肌动蛋白类 基因表达 核酸类 分析 

分 类 号：R512.3[医药卫生—内科学]