Dominant Expression And Antigenicity Detection Of Parvovirus B19 VP2 Protein

Objectives:There is a very close relationship between human parvovirus B19 and many kinds of mankind diseases. Human parvovirus B19(B19) can result in idiopathic thrombocytopenic purpura, acute aplastic anemia, juvenile rheumatoid arthritis, allergic purpura, and other diseases. Parvovirus B19 is also one of the etiopathogenisis for hydrops fetalis, fetal death, abortion and congenital deformity. Two structural proteins (VP1, 85kDa and VP2, 58kDa) and a non-structural protein are encoded by parvovirus B19 genome. The human body can generate specificity antibody after stimulated by VP1 and VP2. So VP1 and VP2 are available for diagnosing B19 infection. B19-VP2-IgM is suitable for early diagnosing parvovirus B19 infection and can be detected in the blood 3 days later after human was infected, and reach a high level in 2 or 3 weeks. B19-VP2-IgM will remain a high level in the body for 2 or 3 months. However, few serology agents for detecting B19 were developed in our country although it is very important for diagnosis of many diseases. So it is necessary to study and develop a new kind of B19-VP2 detection agent which is both affordable and technologically feasible for clinical using.By using molecular biology technology, many viral capsid proteins can be expressed in vitro. In this study, a DNA fragment (4354-4662bp) which had a higher availability of surface in VP2 protein was selected according to the sequence of B19 genome. This gene fragment was obtained by using molecular biology techniques, and some biological functions of the expressed target protein were investigated by ELISA. Methods and results:1. Cloning and sequencing of selected B19-VP2 gene fragment:With the designed primers, a gene fragment (309bp) was amplified by PCR and then inserted into the cloning vector pMD18-T. The sequencing result showed that the...