Diagnostic significance of immunoglobulin G avidity in symptomatic and asymptomatic West Nile virus infection

Abstract INTRODUCTION West Nile virus (WNV) immunoglobulin M (IgM) antibodies have been shown to persist for up to 500 days in certain patients. To evaluate the usefulness of immunoglobulin G (IgG) avidity assessment in the diagnosis of WNV infection, we analyzed 54 WNV IgM- and/or IgG-positive serum samples from 39 patients with neuroinvasive disease and 15 asymptomatic cases tested during a seroprevalence investigation. METHODS Serological tests (WNV IgM/IgG antibody detection, IgG avidity) were performed using commercially available enzyme-linked immunosorbent assays. RESULTS WNV IgM antibodies were detected in 47 (87%) samples. Acute/recent WNV infection was confirmed based on low/borderline avidity index (AI) in 44 IgM-positive samples (93.6%). In three IgM-positive samples (6.4%), high IgG AIs were detected, thus indicating persisting IgM antibodies from previous infections. All IgM-negative samples showed high AIs. Patients with WNV neuroinvasive disease tested within 30 days showed low AIs. In six patients tested 34-50 days after disease onset, AI was borderline (42%-60%), suggesting earlier WNV IgG maturation. Samples with the highest IgM values were associated with the lowest AIs (Spearman's rho coefficient -0.767, p < 0.001). CONCLUSIONS Our results indicate that IgG avidity differentiates current/recent WNV infection from persistent IgM seropositivity from the previous WNV transmission season both in patients with WNV neuroinvasive disease and in asymptomatic persons. A strong negative correlation between IgM antibody levels and AI indicates that in cases with very high IgM levels, determination of IgG avidity may not be necessary. As many patients showed rapid avidity maturation, low IgG avidity is indicative of WNV infection within the previous month.

The First Reported Case of West Nile Encephalitis in Korea

West Nile encephalitis was first identified in 1937, but until now, it was never diagnosed in Korea. A 58-yr-old Korean man was admitted with headache and cognitive dysfunction. The patient had been on a business trip in Guinea. Cerebrospinal fluid (CSF) showed pleocytosis. The patient complained of both leg weakness,and arachnoiditis and myelitis were observed on lumbar magnetic resonance imaging (MRI). A specific neutralizing antibody for West Nile virus was positive in serum. After a treatment with interferon-alpha 3mu, follow up CSF findings recovered completely after 3 months later. The first case of West Nile encephalitis in Korea was imported from Guinea, and was cured successfully.

The First Reported Case of West Nile Encephalitis in Korea

West Nile encephalitis was first identified in 1937, but until now, it was never diagnosed in Korea. A 58-yr-old Korean man was admitted with headache and cognitive dysfunction. The patient had been on a business trip in Guinea. Cerebrospinal fluid (CSF) showed pleocytosis. The patient complained of both leg weakness,and arachnoiditis and myelitis were observed on lumbar magnetic resonance imaging (MRI). A specific neutralizing antibody for West Nile virus was positive in serum. After a treatment with interferon-alpha 3mu, follow up CSF findings recovered completely after 3 months later. The first case of West Nile encephalitis in Korea was imported from Guinea, and was cured successfully.

Production and diagnostic application of recombinant domain III of West Nile envelope protein in Brazil

INTRODUCTION: West Nile virus (WNV) is a flavivirus with a natural cycle involving mosquitoes and birds. Over the last 11 years, WNV has spread throughout the Americas with the imminent risk of its introduction in Brazil. METHODS: Envelope protein domain III of WNV (rDIII) was bacterially expressed and purified. An enzyme-linked immunosorbent assay with WNV rDIII antigen was standardized against mouse immune fluids (MIAFs) of different flavivirus. RESULTS: WNV rDIII reacted strongly with St. Louis encephalitis virus (SLEV) MIAF but not with other flaviviruses. CONCLUSIONS: This antigen may be a potentially useful tool for serologic diagnosis and may contribute in future epidemiological surveillance of WNV infections in Brazil.

Serological detection of West Nile virus in horses and chicken from Pantanal, Brazil

In an effort to detect West Nile virus (WNV) in Brazil, we sampled serum from horses and chickens from the Pantanal region of the state of Mato Grosso and tested for flavivirus-reactive antibodies by blocking ELISA. The positive samples were further confirmed for serological evidence of WNV infection in three (8%) of the 38 horses and one (3.2%) of the 31 chickens using an 80% plaque-reduction neutralisation test (PRNT80). These results provide evidence of the circulation of WNV in chickens and horses in Pantanal.

Serological detection of St. Louis encephalitis virus and West Nile virus in equines from Santa Fe, Argentina

St. Louis encephalitis virus (SLEV) and West Nile virus (WNV) present ecological and antigenic similarities and are responsible for serious human diseases. In addition, WNV is a significant pathogen in terms of equine health. The purpose of our study was to analyse the seroprevalence of SLEV and WNV in equine sera collected in Santa Fe Province, Argentina. The seroprevalence determined using the plaque reduction neutralisation test was 12.2% for SLEV, 16.2% for WNV and 48.6% for a combination of both viruses. These results provide evidence of the co-circulation of SLEV and WNV in equines in Santa Fe.

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.

Seroconversion for west Nile and St. Louis encephalitis viruses among sentinel horses in Colombia

We prospectively sampled flavivirus-naïve horses in northern Colombia to detect West Nile virus (WNV) and St. Louis encephalitis virus (SLEV) seroconversion events, which would indicate the current circulation of these viruses. Overall, 331 (34.1 percent) of the 971 horses screened were positive for past infection with flaviviruses upon initial sampling in July 2006. During the 12-month study from July 2006-June 2007, 33 WNV seroconversions and 14 SLEV seroconversions were detected, most of which occurred in the department of Bolivar. The seroconversion rates of horses in Bolivar for the period of March-June 2007 reached 12.4 percent for WNV and 6.7 percent for SLEV. These results comprise the first serologic evidence of SLEV circulation in Colombia. None of the horses sampled developed symptoms of encephalitis within three years of initial sampling. Using seroconversions in sentinel horses, we demonstrated an active circulation of WNV and SLEV in northern Colombia, particularly in the department of Bolivar. The absence of WNV-attributed equine or human disease in Colombia and elsewhere in the Caribbean Basin remains a topic of debate and speculation.

Neutralising antibodies for West Nile virus in horses from Brazilian Pantanal

Despite evidence of West Nile virus (WNV) activity in Colombia, Venezuela and Argentina, this virus has not been reported in most South American countries. In February 2009, we commenced an investigation for WNV in mosquitoes, horses and caimans from the Pantanal, Central-West Brazil. The sera of 168 horses and 30 caimans were initially tested using a flaviviruses-specific epitope-blocking enzyme-linked immunosorbent assay (blocking ELISA) for the detection of flavivirus-reactive antibodies. The seropositive samples were further tested using a plaque-reduction neutralisation test (PRNT90) for WNV and its most closely-related flaviviruses that circulate in Brazil to confirm the detection of specific virus-neutralising antibodies. Of the 93 (55.4 percent) blocking ELISA-seropositive horse serum samples, five (3 percent) were seropositive for WNV, nine (5.4 percent) were seropositive for St. Louis encephalitis virus, 18 (10.7 percent) were seropositive for Ilheus virus, three (1.8 percent) were seropositive for Cacipacore virus and none were seropositive for Rocio virus using PRNT90, with a criteria of > four-fold antibody titre difference. All caimans were negative for flaviviruses-specific antibodies using the blocking ELISA. No virus genome was detected from caiman blood or mosquito samples. The present study is the first report of confirmed serological evidence of WNV activity in Brazil.

Is West Nile virus a potential cause of central nervous system infection in Brazil? / Seria o vírus do Oeste do Nilo causa potencial de infecção no sistema nervoso central no Brasil?

Meningitis and encephalitis are complications of West Nile virus (WNV) infection. Although WNV is endemic in North America, the virus has recently been reported in Colombia and Argentina. Investigation of WNV in Brazil is important since this virus has never been studied previously in this country. OBJECTIVE: To investigate the presence of WNV infection in viral encephalitis/meningitis cases of unknown etiology in the city of Rio de Janeiro, Brazil. METHOD: Thirty-seven adults with viral meningitis/encephalitis had their serum and CSF tested for WNV antibodies using the ELISA method. RESULTS: Only one case was WNV-positive, but this case was also positive for dengue. The plaque reduction neutralization test distinguished infections, and was negative for WNV. CONCLUSION: WNV can be confused with dengue infection. Their symptoms and neurological picture are similar. We did not find WNV in any patients with encephalitis and meningitis in the city of Rio de Janeiro. Up to now, it has not been detected in Brazil.

Meningite e encefalite são complicações da infecção pelo vírus do Oeste do Nilo (VON). Embora o VON seja endêmico na América do Norte, recentemente o vírus foi descrito na Colômbia e Argentina. Sua pesquisa no Brasil é importante uma vez que o vírus nunca fora estudado antes em nosso país. OBJETIVO: Investigar a presença do VON em casos de meningite e encefalite viral de etiologia desconhecida, na cidade no Rio de Janeiro, Brasil. MéTODO: Trinta e sete adultos com quadro de meningite/encefalite tiveram seu LCR e soro testados para anticorpos anti-VON, pelo método ELISA. RESULTADOS: Apenas um caso obteve sorologia positiva para VON, mas a sorologia para dengue também fora positiva. O teste da neutralização por redução de placa foi utilizado para distinção entre as infecções, sendo negativo para VON. CONCLUSÃO: A infecção por VON pode ser confundida com a infecção pelo vírus da dengue, seus sintomas e quadro neurológico são similares. Nós não encontramos o VON em pacientes com meningite/ encefalite na cidade do Rio de Janeiro. Até o momento ele ainda não foi identificado no Brasil.

Prevalencia de infección por el virus del Nilo Occidental en dos zoológicos del estado de Tabasco / Prevalence of West Nile Virus infection in animals from two state zoos Tabasco

OBJETIVO: Determinar la prevalencia de infección por el virus del Nilo Occidental (VNO) en animales, mosquitos y personal que labora en dos zoológicos del estado de Tabasco, en México. MATERIAL Y MÉTODOS: Con la utilización de ELISA de bloqueo se detectaron anticuerpos en sueros de animales: se buscó un fragmento del genoma del VNO por RT-PCR en el suero de animales, empleados y mosquitos. RESULTADOS: En el zoológico "La Venta" se encontró una seroprevalencia de 25.67 por ciento (19/74) en aves y de 85.71 por ciento (6/7) en reptiles. En el zoológico "Yum-Ká", 31.25 por ciento (50/160) de las aves y 34.48 por ciento (16/29,) de los mamíferos, tuvieron anticuerpos contra el VNO. En un grupo de mosquitos (Culex quinquefasciatus) se detectó el genoma del virus. CONCLUSIONES: La detección de anticuerpos contra el VNO en animales de ambos zoológicos y del genoma viral en mosquitos demuestra la presencia del virus, lo cual representa un riesgo potencial de infección para animales y humanos.

OBJECTIVE: To determine the prevalence of West Nile Virus (WNV) infection in animals, mosquitoes and employees from two zoos of Tabasco state, Mexico. MATERIAL AND METHODS: WNV antibodies were detected by blocking ELISA in serum samples from animals. Viral RNA was detected by RT-PCR from mosquitoes and serum samples from employees at "Yum-Ká" zoo. RESULTS: Seroprevalence in birds was 25.65 percent (19/74) and 85 percent (6/7) in reptiles from "La Venta" zoo. Thirty-one percent of birds (50/160) and 34.48 percent mammals (16/29) at the "Yum-Ká" zoo, were seropositive. All human serum samples from Yum-ká zoo were negative by RT-PCR. A pool of mosquitoes (Culex quinquefasciatus) was positive for WNV. CONCLUSIONS: The presence of WNV antibodies in animals from both zoos and the detection of viral genome in mosquitoes demonstrate the presence of WNV in this region and indicates a potential risk of infection in animals and humans.

West Nile virus activity in Latin America and the Caribbean

OBJETIVOS: El virus del Nilo occidental (VNO, familia Flaviviridae, género Flavivirus) se ha propagado rápidamente por toda la cuenca del Caribe desde que se detectó por primera vez en 2001. En este informe se resumen nuestros conocimientos actuales acerca de la transmisión del VNO en zonas tropicales del continente americano. MÉTODOS: Revisamos todo lo que se ha publicado sobre el tema y consultamos a autoridades de salud clave para obtener datos inéditos. RESULTADOS: Las infecciones por el virus del Nilo occidental aparecieron por primera vez en seres humanos residentes de las Islas Caimán y de los Cayos de la Florida en 2001, y en pájaros de aspecto sano de los cuales se obtuvieron muestras a principios de 2002. En 2002 se encontraron pruebas serológicas de infección por el VNO en caballos, pollos y aves de corral no estabuladas oriundas de Guadalupe, la República Dominicana y la parte oriental de México. En 2003, el VNO se diseminó dentro de México y por la parte norte de Centroamérica y se encontraron pruebas serológicas en las Bahamas, Puerto Rico y Cuba. En 2004, las primeras pruebas serológicas de actividad vírica en ecosistemas sudamericanos se detectaron en septiembre y octubre en Colombia y Trinidad, donde se observaron anticuerpos neutralizantes contra el VNO en animales domésticos. CONCLUSIONES: Estos informes esporádicos de enfermedad equina, humana y aviar en América Latina y el Caribe son desconcertantes. Es necesario aislar las cepas para determinar si la atenuación del virus u otro factor explica la carga de enfermedad reducida en ecosistemas tropicales.

A serological survey of arboviral diseases among the human population of the Andaman and Nicobar Islands, India.

In an attempt to determine the prevalence of certain arthropod-borne viruses of public health importance amongst the human population of the Andaman and Nicobar Islands of India, 2,401 sera were collected from six major localities. The sera were analysed by the hemagglutination inhibition (HI) and neutralization (N) tests, using Chikungunya (CHIK), Japanese encephalitis (JE), West Nile (WN), dengue (DEN-2), Langat (TP-21) and Kyasanur Forest disease (KFD) viral antigens. The highest prevalence of HI antibodies was detected against KFD virus (22.4%), followed by Langat (20.2%), JE (5.9%), DEN-2 (3.1%), CHIK (2.9%) and WN (0.8%) viruses. Cross-reactions to the viral antigens were also noted. The results of N tests indicated a high prevalence of DEN-2 (25.4%) virus, followed by Langat (17.5%), CHIK (15.3%), KFD (12%), JE (2.19%) and WN (1.8%). These results are discussed in relation to important epidemiological parameters like age, sex and geographical location. To our knowledge, this is the first report of an extensive serosurvey of arthropod-borne viruses on these islands.

Prevalence of West Nile virus infection in India.

During the course of the virological investigation of cases of suspected viral fevers carried out at the National Institute of Virology (NIV), Pune, India, evidence of recent infection with West Nile (WN) virus was detected in 88 cases. Fever, general aches, headache, nausea and vomiting were the principal clinical features in 92% (81/88) of the cases; there were seven cases of encephalitis, in which WN virus-specific IgM class antibodies were detected in CSF samples. These cases of encephalitis were from Japanese encephalitis (JE) nonendemic areas, like Maharashtra and Rajasthan, as well as from JE endemic areas, like Goa and Orissa. Interestingly, neutralizing antibodies predominantly to WN virus were detected in CSF samples by the 50% cytopathic effect inhibition method; the titers ranged from 5 to 375. Cases of WN virus infection associated with both encephalitis and classic features have been reported for the first time in recent years in India. Reports of unique urban West Nile virus encephalitis epidemics in New York, Romania, and Algeria in recent years have signaled the emergence of neurological infection due to West Nile virus as a novel public health threat. This study is important because it records evidence of WN virus infection in India.

Antigen distribution pattern of West Nile virus in Culex tritaeniorhynchus, Culex vishnui and Culex pseudovishnui mosquitoes.

Distribution of West Nile (WN) virus antigen in different tissues of mosquitoes was studied in three species viz., Culex tritaeniorhynchus, C. vishnui and C. pseudovishnui. Overall per cent positivity was higher in the intra thoracically inoculated as compared to the orally infected mosquitoes, suggesting the existence of a midgut barrier. In a small number of mosquitoes salivary glands were found negative even though fluorescence was seen in the respective head squashes, suggesting salivary gland barrier in these mosquitoes. There was no difference in the per cent salivary gland and salivary gland area positivity between these three species. Presence of virus antigen in the ovaries of these three species on the 3rd post infection day suggests the possibility of transovarial transmission of virus even in the first gonotrophic cycle, which is of epidemiological importance.

Prevalence of Japanese encephalitis and West Nile viral infections in pig population in and around Chandigarh.

A study on sero prevalence of Japanese Encephalitis (JE) and West Nile (WN) viruses in pigs was carried out in and around Chandigarh. A total of 158 blood samples collected (June, 1995-March, 1996) from batches of pigs were screened for antibodies against JE and WN viral antigens by haemagglutination inhibition and complement fixation tests. In Chandigarh out of 13 pigs, 5 had JE and one WN HAI antibodies. Similarly, the sera of 14 and 1 pigs in Punjab and 29 and 3 pigs in Haryana were positive for JE and WN HAI antibodies respectively. In total JE specific HAI antibodies were found in 30.3% and CF antibodies in 12.5% of pigs whereas WN specific HAI antibodies were obtained in 3.2% and only one pig had CF antibodies. The results show that JE and WN viral infections are prevalent in pig population in and around Chandigarh.

Post-epidemic serological survey for JE virus antibodies in south Arcot district (Tamil Nadu).

Following an extensive epidemic of Japanese encephalitis (JE) in 1981, a serological survey was carried out in the South Arcot district of Tamil Nadu in April 1982. Serum specimens were obtained from 10 villages with serologically proven infections and 6 villages with no reported case of encephalitis during the 1981 epidemic. The serum specimens tested for haemagglutination inhibition, complement fixing and neutralizing antibodies, showed a high prevalence of JE (49.17%), West Nile (40.78%) and dengue (18.14%) viruses with predominance of JE. The prevalence did not differ between the villages with or without Japanese encephalities. There was also no difference between the family members and neighbours.