MOLECULAR DIAGNOSTIC TESTS FOR SIMIAN HERPESVIRUS

点击翻译按钮获取中文摘要

Monkeys, particularly rhesus macaques, are a vital resource for biomedical research. Herpesvirus simiae (B virus), a natural pathogen of macaques, presents a very real danger for biomedical personnel who work with these animals. Sensitive assays for detection of infected monkeys are needed to identify animals which pose a risk to personnel and to identify B virus-free animals for use in establishment of B virus-free breeding colonies. H. cercopithicus 1 (SA8) causes problems in baboon breeding colonies. The incidence of transmission of SA8 to humans is unknown. Sensitive assays which can differentiate between B virus, SA8 and the two human herpes simplex viruses (HSVs) are necessary for accurate and rapid identification of human simian herpesvirus infections. The extensive antigenic relatedness of the primate herpesviruses has been a major impediment to achieving this goal. Building on data generated from molecular research in our laboratory, we will develop sensitive ELISA tests for rapid detection and differentiation of primate viruses and antibody to B virus and SA8. These tests will be based on specific viral glycoproteins. A PCR assay will be developed for detection of primate herpesviruses in clinical samples. Using a primer set located in conserved regions of the glycoprotein gB gene, a virus-specific region of the gene will be amplified from any of the four primate herpesviruses. Subsequent analysis of the PCR product will enable determination of which virus (HSV1, HSV2, B virus or SA8) is present in the clinical samples. ELISAs will also be developed for serological detection of suspected human and simian B virus and\or SA8 cases. The gG glycoprotein homologs of all four viruses are antigenically virus-specific. The gG genes of SA8 and BV will be cloned, sequenced and stably expressed in mammalian cells. These cell lines and anti-gG antibodies will be used to develop sensitive, virus-specific ELISAs for detection of antibody to BV and SA8. This project applies results of basic molecular research to the clinically important problem of B virus and SA8 detection and diagnosis. The assays developed will permit rapid, sensitive and reliable differential diagnosis of primate herpesvirus infections. This will enable primate colony managers to more effectively operate primate facilities and reduce loss of human life due to zoonotic herpes infections.