Detection Of The Middle Fragment Derived From ORF â…  Of Borna Disease Virus

Objective: Borna disease virus (BDV) is a neurotropic nonsegmented negative-stranded RNA virus that caused the illness calling Borna disease (BD). The illness is a nervous syndrome characterized by profound behavirol and consciousness abnormalities as well as encephalitis. BDV contanins a 8.9Kb antigenome that exists of six open reading frames correspond to encoding the nucleoprotein (p40), the phosphoprotein (p24), the unglycoslated protein (p10), the glycoslated matrixprotein (p16, p18), the glycoprotein (p57, gp94), and the L-polymerase with a molecular mass of approximately 180kDa. At present we usually use detecting BDV ORF II genome to cofirm whether the people have a BDV infection or not,the research suggests that BDV ORF II is more conservative and less variability,thus it is a good marker for detecting BDV infection. But it is doubt that the speciality and stability for the outcome of detecting BDV ORF I genome.We will detect the indviduals'(include the neuropsychiatric4patients with BDV ORFII postive or not and healthy blood donator) middle fragment of BDV ORF I to help defmiting whether internal healthy people and the certain neuropsychiatric patient such as with viral encephalitis,multiple,schizophrenia,affective disorder have catch a BDV infection or not, most of all, the research is for understanding if there is any difference in the fragment from animal and human beings infected with BDV, thereby to find another sensitive and stably marker for confirming BDV infection, and establish a solide bottom for advanced study.Methods: Extracting the total RNA of human PBMC, the objective include 60 healthy blood donator, 30 patient with viral encephalitis and multiple sclerosis and Parkinsonian syndrome, 30 patient with schizophrenia and affective disorder, this indviuals were inpatients or outpatients of the first hospital of Chongqing University of Medical Science from December, 2000 to June, 2001. Using nested RT-PCR techique to detect Borna disease virus' middle fragment in ORF I, and using southern blot hybridization to analyze the PCR product.Results: 1. The electrophoresis results of PCR product: one of the 20 patient with viral encephalitis had BDV positive (5%), two of the 17 patient with schizophrenia had BDV positive (11.8%), one of the 13 patient with affective disorder had BDV positive (7.7%), none of healthy blood donatorshad BDV positive (0.0%). The positive results have a marked discrepancy between the patients with schizophrenia and the healthy blood donators(P< 0.05).2. The analysis results of southern blot hybridization among all of specimens are coincidence with the detecting results of PCR products electrophoresis.Conclusions: 1. The middle fragment derived from ORF I of Borna disease virus which infected human being and animal has a very high conversation and concordance.2. Detection the middle fragment derived from ORF I of Borna disease virus is another probably sensitive and stably marker for confirming Borna disease virus infection.3.There is a Borna diseae virus infection in the certain neuropsychiatric patients of China.