High Seroprevalence of Jamestown Canyon Virus among Deer and Humans, Nova Scotia, Canada.

Using residual serum samples from Nova Scotia, Canada, we found that 87.8% of tested deer and an estimated 20.6% of the human population were infected with Jamestown Canyon virus. Human seropositivity reached 48.2% in 1 region. This virus may be an underrecognized cause of disease in Nova Scotia.

Evaluation of 5 Commercially Available Zika Virus Immunoassays.

Because of the global spread of Zika virus, accurate and high-throughput diagnostic immunoassays are needed. We compared the sensitivity and specificity of 5 commercially available Zika virus serologic assays to the recommended protocol of Zika virus IgM-capture ELISA and plaque-reduction neutralization tests. Most commercial immunoassays showed low sensitivity, which can be increased.

The memory T cell response to West Nile virus in symptomatic humans following natural infection is not influenced by age and is dominated by a restricted set of CD8+ T cell epitopes.

We examined the West Nile virus (WNV)-specific T cell response in a cohort of 52 patients with symptomatic WNV infections, including neuroinvasive and non-invasive disease. Although all virus proteins were shown to contain T cell epitopes, certain proteins, such as E, were more commonly targeted by the T cell response. Most patients exhibited reactivity toward 3-4 individual WNV peptides; however, several patients exhibited reactivity toward >10 individual peptides. The relative hierarchy of T cell reactivities in all patients showed a fixed pattern that was sustained throughout the 12-mo period of the current study. Surprisingly, we did not observe any relationship between age and either the breadth or magnitude of the T cell response following infection. We also did not observe a relationship between disease severity and either the breadth or magnitude of the T cell response. The T cell epitopes were distributed in a non-random fashion across the viral polyprotein and a limited number of epitopes appeared to dominate the CD8(+) T cell response within our cohort. These data provide important new insight into the T cell response against WNV in humans.

Molecular characterization of a panel of murine monoclonal antibodies specific for the SARS-coronavirus.

The availability of monoclonal antibodies (mAbs) specific for the SARS-coronavirus (SARS-CoV) is important for the development of both diagnostic tools and treatment of infection. A molecular characterization of nine monoclonal antibodies raised in immune mice, using highly purified, inactivated SARS-CoV as the inoculating antigen, is presented in this report. These antibodies are specific for numerous viral protein targets, and six of them are able to effectively neutralize SARS-CoV in vitro, including one with a neutralizing titre of 0.075 nM. A phylogenetic analysis of the heavy and light chain sequences reveals that the mAbs share considerable homology. The majority of the heavy chains belong to a single Ig germline V-gene family, while considerably more sequence variation is evident in the light chain sequences. These analyses demonstrate that neutralization ability can be correlated with specific murine V(H)-gene alleles. For instance, one evident trend is high sequence conservation in the V(H) chains of the neutralizing mAbs, particularly in CDR-1 and CDR-2. The results suggest that optimization of murine mAbs for neutralization of SARS-CoV infection will likely be possible, and will aid in the development of diagnostic tools and passive treatments for SARS-CoV infection.

Development and characterisation of neutralising monoclonal antibody to the SARS-coronavirus.

There is a global need to elucidate protective antigens expressed by the SARS-coronavirus (SARS-CoV). Monoclonal antibody reagents that recognise specific antigens on SARS-CoV are needed urgently. In this report, the development and immunochemical characterisation of a panel of murine monoclonal antibodies (mAbs) against the SARS-CoV is presented, based upon their specificity, binding requirements, and biological activity. Initial screening by ELISA, using highly purified virus as the coating antigen, resulted in the selection of 103 mAbs to the SARS virus. Subsequent screening steps reduced this panel to seventeen IgG mAbs. A single mAb, F26G15, is specific for the nucleoprotein as seen in Western immunoblot while five other mAbs react with the Spike protein. Two of these Spike-specific mAbs demonstrate the ability to neutralise SARS-CoV in vitro while another four Western immunoblot-negative mAbs also neutralise the virus. The utility of these mAbs for diagnostic development is demonstrated. Antibody from convalescent SARS patients, but not normal human serum, is also shown to specifically compete off binding of mAbs to whole SARS-CoV. These studies highlight the importance of using standardised assays and reagents. These mAbs will be useful for the development of diagnostic tests, studies of SARS-CoV pathogenesis and vaccine development.

Comparison of assays for the detection of West Nile virus antibodies in chicken serum.

Six tests for the detection of West Nile virus (WNV) antibodies in the serum of experimentally infected chickens were compared. The tests included the hemagglutination-inhibition test (HIT), immunoglobulin M (IgM)-capture enzyme-linked immunosorbent assay (ELISA) with WNV-infected mouse brain antigen, immunoglobulin G (IgG) indirect ELISA with tickborne encephalitis viral antigen, the microtitre virus neutralization test, the standard plaque reduction neutralization test (PRNT), and the microtitre PRNT (micro-PRNT). Thirty adult chickens, intravenously and intramuscularly inoculated with 10(7) plaque-forming units (PFU) of WNV strain Egypt 101, were bled and given a booster of 10(7) PFU at 7,15, and 21 d postinoculation; the final blood collection was on day 28. Although the micro-PRNT is capable of detecting the highest antibody titres during both early and late infection, because of the technical complexity and time requirements of this test a combination of IgM and IgG ELISAs is recommended for serologic screening. Serum samples that give positive results in the ELISAs can then be tested by the micro-PRNT to determine the specificity of antibodies to WNV.

Evaluating the use of house sparrow nestlings as sentinels for West Nile virus in Saskatchewan.

This study evaluated the use of house sparrow (Passer domesticus) nestlings as sentinels of West Nile virus (WNV) in the prairie grasslands of Saskatchewan. In the summer of 2006, 600 house sparrow nestlings were collected and pooled tissues tested by reverse transcriptase-polymerase chain reaction. All tested negative for WNV. During the same period, no WNV was detected by mosquito surveillance in the study area and 15 WNV-infected pools were collected from the nearby city of Estevan. Six percent of avian carcasses collected from Regina, a city 100 km from the study area in the same ecozone, were infected with WNV. In 2007, 200 house sparrow nestlings were collected and 4 tested positive for WNV, a prevalence of 2%. Ninety-seven house sparrow eggs were also collected and WNV antibodies were measured in the yolk. Seven eggs had measurable titers, a prevalence of 7.2%. Combined WNV surveillance showed high levels of WNV transmission in 2007; 112 WNV-infected mosquito pools were collected from nearby cities of Estevan and Weyburn, and the proportion of WNV infected avian carcasses from Regina was 78%. There were 1456 human cases of WNV in Saskatchewan in 2007, compared to 19 cases in 2006. The study concluded that house sparrow nestlings are not useful as an early warning of WNV circulation, or as a measure of the intensity of WNV activity in the prairie grasslands. Also, the study determined that maternally derived antibody did not have a significant limiting effect on WNV transmission to house sparrow nestlings in 2007, a year of epidemic WNV activity in the study area.

[Serological study carried out in Cuban localities where confirmed western Nile virus infection is present]. / Estudio serológico en localidades cubanas con infecciones confirmadas al virus del Nilo Occidental.

INTRODUCTION: first infected cases caused by West Nile virus were reported in Cuba in 2004. OBJECTIVE: to monitor and learn about the prevalence of the West Nile virus in those areas with confirmed cases. METHODS: the study was conducted in Jatibonico municipality and in the city of sancti Spiritus. A total number of 14 persons, 8 horses and 41 birds were researched to detect antibodies to flavivirus and specific antibodies to West Nile virus. RESULTS: the presence of specific antibodies to West Nile virus was confirmed in 4 samples of sera from birds and in 4 from horses. One person was confirmed as one case of asymptomatic West Nile virus infection. CONCLUSIONS: the presence of specific antibodies to West Nile virus in birds, horses and persons residing in areas where there are confirmed cases showed that a local amplification cycle had been established in Cuba before this study.

Seroprevalence of 10 zoonotic infections in 2 Canadian Cree communities.

We evaluated the seroprevalence of 10 zoonotic agents among the general population (15 years old and over) of Eastmain and Wemindji, James Bay, Quebec, in 2007. Overall seroprevalence rates were similar between the 2 communities. Nearly half the individuals tested (n = 251; 146 women, 105 men) were seropositive (n = 115) for at least one zoonosis. The highest seroprevalence rates were for Leptospira sp. (23%), Francisella tularensis (17%), and the California serogroup viruses (JC and SSH viruses) (10%). The other zoonoses (Toxoplasma gondii, Coxiella burnetii, Echinococcus granulosus, Toxocara canis, and Trichinella sp.) had seroprevalence rates ≤5%; no exposures were identified to hantaviruses (Sin Nombre virus). Overall, seropositivity was related to age, gender, hunting, and owning a dog. There was no medical history suggestive of overt diseases. Nonetheless, physicians should consider these agents when confronted with difficult or confusing diagnoses. In particular, the bacterial zoonoses should be ruled out in individuals with high or prolonged fever.

Estudio serológico en localidades cubanas con infecciones confirmadas al virus del Nilo Occidental / Serological study carried out in Cuban localities where confirmed Western Nile virus infection is present

Introducción: las primeras infecciones por el virus del Nilo Occidental en Cuba se reportaron en 2004. Objetivo: monitorear y conocer la prevalencia del virus del Nilo Occidental en áreas con casos confirmados de este. Métodos: el estudio se llevó a cabo en la municipalidad de Jatibonico y en la ciudad de Sancti Spiritus. Un total de 14 personas, 8 caballos y 41 aves se estudiaron para la detección de anticuerpos a flavivirus y específicos al virus del Nilo Occidental. Resultados: se confirmó la presencia de anticuerpos específicos a virus del Nilo Occidental en 4 muestras de suero de aves y 4 de caballos. Una persona se confirmó como 1 caso de infección por virus del Nilo Occidental asintomático. Conclusiones: la presencia de anticuerpos específicos al virus del Nilo Occidental en aves residentes, caballos y humanos en áreas con casos confirmados demuestran el establecimiento de un ciclo de amplificación local establecido en Cuba antes de este estudio.

Introduction: First infected cases caused by West Nile virus were reported in Cuba in 2004. Objective: to monitor and learn about the prevalence of the West Nile virus in those areas with confirmed cases. Methods: the study was conducted in Jatibonico municipality and in the city of Sancti Spiritus. A total number of 14 persons, 8 horses and 41 birds were researched to detect antibodies to flavivirus and specific antibodies to West Nile virus. Results: the presence of specific antibodies to West Nile virus was confirmed in 4 samples of sera from birds and in 4 from horses. One person was confirmed as one case of asymptomatic West Nile virus infection. Conclusions: the presence of specific antibodies to West Nile virus in birds, horses and persons residing in areas where there are confirmed cases showed that a local amplification cycle had been established in Cuba before this study.

West Nile Virus infection in humans and horses, Cuba.

A surveillance system to detect West Nile virus (WNV) was established in Cuba in 2002. WNV infection was confirmed by serologic assays in 4 asymptomatic horses and 3 humans with encephalitis in 2003 and 2004. These results are the first reported evidence of WNV activity in Cuba.

West Nile virus encephalitis in a Barbary macaque (Macaca sylvanus).

An aged Barbary ape (Macaca sylvanus) at the Toronto Zoo became infected with naturally acquired West Nile virus encephalitis that caused neurologic signs, which, associated with other medical problems, led to euthanasia. The diagnosis was based on immunohistochemical assay of brain lesions, reverse transcriptase-polymerase chain reaction, and virus isolation.

Rapid antigen-capture assay to detect West Nile virus in dead corvids.

The utility of the VecTest antigen-capture assay to detect West Nile virus (WNV) in field-collected dead corvids was evaluated in Manitoba and Ontario, Canada, in 2001 and 2002. Swabs were taken from the oropharynx, cloaca, or both of 109 American Crows, 31 Blue Jays, 6 Common Ravens, and 4 Black-billed Magpies from Manitoba, and 255 American Crows and 28 Blue Jays from Ontario. The sensitivity and specificity of the antigen-capture assay were greatest for samples from American Crows; oropharyngeal swabs were more sensitive than cloacal swabs, and interlaboratory variation in the results was minimal. The sensitivity and specificity of the VecTest using oropharyngeal swabs from crows were 83.9% and 93.6%, respectively, for Manitoba samples and 83.3% and 95.8%, respectively, for Ontario birds. The VecTest antigen-capture assay on oropharyngeal secretions from crows is a reliable and rapid diagnostic test that appears suitable for incorporation into a WNV surveillance program.