Arrand. antigen-based ELISA and Western blotting. The diagnostic sensitivities of the gB-, gC-, gD-, and mgG-ELISAs were 100, 97.3, 88.0, and 80.0%, respectively. The specificities of the gB-, gC-, and gD-ELISAs and of the mgG-ELISA were 100 and 97.5%, respectively. In contrast, the sensitivities and specificities of sgG- and gE-ELISAs were low, suggesting that sgG and gE are less effective diagnostic antigens. Sera from nonmacaque monkeys cross-reacted with gB, gC, and gD, and only baboon sera reacted weakly with mgG. Human herpes simplex virus type 1 (HSV-1)- and HSV-2-positive sera pools reacted with gB and gD, whereas sera from B virus-infected individuals reacted with all four antigens. These data show that gB, gC, gD, and mgG have a high diagnostic potential for B computer virus serodiagnosis in macaques, whereas mgG may be a valuable antigen for discrimination between antibodies induced by B computer virus and those induced by other, closely related alphaherpesviruses, including HSV-1 and -2. Human contamination with B computer virus (also called cercopithecine herpesvirus 1, monkey B computer virus, and herpes B computer virus) is the most feared occupational hazard among individuals working with macaque monkeys, since fatality is usually often the end result of contamination, which proceeds in the absence of effective antiviral therapy (25, 56). The use Efaproxiral of macaques in research has been steadily growing over the last decade and is expected to rise quickly in the near future due to the increasing demands for these animals for use in HIV/AIDS investigations, vaccine trials, drug testing, and research into bioterrorism agents. As macaque usage increases, frequencies of human exposures to B virus are increasing as well. Rapid and accurate diagnostic tests are urgently needed to aid in the early identification of clinical cases, which is essential for a timely initiation of antiviral therapies in zoonotically infected humans. In addition to human diagnostics, enhanced assays are required for monitoring specific-pathogen-free (SPF) macaque colonies established by the National Institutes of Health for the breeding of B virus-free animals (55), as these animals often demonstrate only very low levels of antibody. Unfortunately, a direct diagnosis of infection by virus detection (cell culture Efaproxiral or PCR) is impossible in most cases, since, similar to other alphaherpesviruses, B virus establishes a lifelong latency in sensory ganglia of macaques and seldom reactivates (9, 53, 58). Current diagnoses of B virus infections in humans and monkeys rely mainly on the detection of serum antibodies to B virus proteins. Indirect enzyme-linked immunosorbent assays (ELISA) and other rapid serological tests based on a solubilized, B virus-infected cell antigen have been developed and used for the identification of infected animals (8, 21, 28, 39), with subsequent confirmation by Western blotting to identify specific targets that are immunoreactive with serum antibodies (54). Serodiagnoses of zoonotic infections are performed by Western blotting, which is a time-consuming technique that requires visual interpretations of complex patterns. This method is not specific enough to unequivocally identify B virus infections in herpes simplex virus (HSV)-positive humans because antibodies to HSV type 1 (HSV-1) and HSV-2 are highly cross-reactive with B virus proteins (12, 23), making differentiation a complex task. Most importantly, currently used serological assays utilize B Efaproxiral virus-infected cell lysates as an antigen, and these can only be produced in a maximum containment laboratory (biosafety level 4), which limits the number of facilities that are capable of providing antigen. Antigens may also suffer from lot-to-lot variation, compromising outcome measures based on assays using these antigens. Recombinant-based serological assays have been developed for the diagnosis of many viral infections, including human cytomegalovirus (11), hepatitis C virus (27), hepatitis E virus (46), human papillomavirus (49), Ebola virus (45), and many others. Several recombinant glycoprotein G (gG)-based immunoassays for HSV type-specific serodiagnosis are commercially available (19, 44). However, the use of recombinant antigens for B virus Rabbit polyclonal to PHYH serodiagnosis has not been widely investigated. Recently, recombinant gD was shown to be useful for B virus serodiagnosis by dot blot and Western blot assays, but the performance of this antigen in ELISAs was not studied (51). In an earlier study, we produced a fusion protein containing a single B virus-specific immunodominant epitope of gD and demonstrated its efficacy for the identification of B virus infections by using an indirect ELISA (43). The serodiagnosis of infections, however, cannot be based exclusively upon the presence of antibodies to a single epitope of a pathogen due to variations in individual responses to a selected epitope. Moreover, the existence of cross-reacting antibodies against similar epitopes in other proteins may result in a false-positive diagnosis. Ideally, the.