scholarly journals Monoclonal Antibody-Based Competitive Enzyme-Linked Immunosorbent Assay for Detecting and Quantifying West Nile Virus-Neutralizing Antibodies in Horse Sera

2006 ◽  
Vol 14 (2) ◽  
pp. 134-138 ◽  
Author(s):  
Kang-Seuk Choi ◽  
Young-Joon Ko ◽  
Jin-Ju Nah ◽  
Yong-Joo Kim ◽  
Shien-Young Kang ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
West Nile Virus ◽  
Immunosorbent Assay ◽  
Neutralizing Antibodies ◽  
Large Scale ◽  
Good Alternative ◽  
Standard Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
West Nile

ABSTRACT A rapid immunoassay for detecting and quantifying West Nile virus (WNV)-neutralizing antibodies in sera was developed as an alternative to the plaque reduction neutralization test (PRNT), the gold standard test for WNV. The assay is a competitive, enzyme-linked immunosorbent assay using neutralizing monoclonal antibody 5E8 (NT-ELISA). A cutoff percent inhibition (PI) value of 35% (mean PI plus 3 standard deviations), with a specificity of 99%, was established based on analysis of 246 serum samples from horses free of WNV. The NT-ELISA detected neutralizing antibodies in all sera collected 7 or 14 days postinoculation from mice (n = 11) infected with lineage I (strain NY385-99) or II (strain B956) WNV. When sera from WNV-vaccinated horses (n = 212) were tested by NT-ELISA and PRNT, the NT-ELISA gave a positive result for 96.1% (173/180) of the PRNT-positive sera and 3.1% (1/32) of the PRNT-negative sera. Discrepancies between the two tests were observed mainly with sera with low PRNT90 titers (expressed as the reciprocal of the highest dilution yielding ≥90% reduction in the number of plaques) for WNV or low PIs by NT-ELISA. The overall agreement (k value) between the two tests was 0.86. A good correlation (r 2 = 0.77) was also observed between the tests for endpoint titration of sera (n = 116). In conclusion, the newly developed NT-ELISA may be a good alternative serologic assay for detecting WNV that can be used for large-scale testing of WNV-neutralizing antibodies in multiple species.

scholarly journals Prevalence of feline coronavirus antibodies in cats in Bursa province, Turkey, by an enzyme-linked immunosorbent assay

2009 ◽  
Vol 11 (10) ◽  
pp. 881-884 ◽  
Author(s):  
Annamaria Pratelli ◽  
Kadir Yesilbag ◽  
Marcello Siniscalchi ◽  
Ebru Yalçm ◽  
Zeki Yilmaz
Keyword(s):  
Immunosorbent Assay ◽  
Predictive Value ◽  
Gold Standard ◽  
Population Based ◽  
Standard Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
High Association ◽  
Gold Standard Test ◽  
Feline Coronavirus

Feline sera from Bursa province (Turkey) were assayed for coronavirus antibody using an enzyme-linked immunosorbent assay (ELISA). The study was performed on 100 sera collected from cats belonging to catteries or community shelters and to households. The serum samples were initially tested with the virus neutralisation (VN) test and the results were then compared with the ELISA. The VN yielded 79 negative and 21 positive sera but the ELISA confirmed only 74 as negative. The ELISA-negative sera were also found to be free of feline coronoviruses-specific antibodies by Western blotting. Using the VN as the gold standard test, ELISA had a sensitivity of 100% and a specificity of 93.6%, with an overall agreement of 95%. The Kappa (κ) test indicated high association between the two tests (κ=0.86, 95% confidence interval (CI) 0.743–0.980). The positive predictive value (PPV) was 0.8, and the negative predictive value (NPV) was 0.93. The prevalence of FCoV II antibodies in the sampled population based on the gold standard was 62% (95% CI 0.44–0.77) among multi-cat environments, and 4% (95% CI 0.01–0.11) among single cat households.

scholarly journals Development of an Enzyme-Linked Immunosorbent Assay for Immunoglobulin M Antibodies against Measles Virus

2003 ◽  
Vol 10 (3) ◽  
pp. 439-442 ◽  
Author(s):  
F. Roodbari ◽  
M. H. Roustai ◽  
A. Mostafaie ◽  
H. Soleimanjdahi ◽  
R. Sarrami Foroshani ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Neutralizing Antibodies ◽  
Measles Virus ◽  
Clinical Symptoms ◽  
Maculopapular Rash ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Vaccination Programs ◽  
Elisa System

ABSTRACT Measles is a highly contagious respiratory virus infection, with typical clinical symptoms including maculopapular rash, fever, cough, coryza, and conjunctivitis. Despite implementation of widespread vaccination programs throughout the world, the rates of global morbidity and mortality are still considerable. This study was performed to design a reliable indirect enzyme-linked immunosorbent assay (ELISA) to measure measles-specific immunoglobulin M (IgM). First, human IgM was purified, and then an anti-IgM antibody was produced in rabbits and purified in a multistep process. The rabbit IgG against human IgM was conjugated with peroxidase. Measles virus-infected Vero cells produced viral antigen. One hundred serum samples from infants of 9 to 18 months of age, mostly vaccinated, were evaluated for determining the presence of specific IgM antibodies against measles virus. The samples were also evaluated for neutralizing antibodies against measles virus by a microneutralization test (MNT). By comparing the results of the ELISA with those of MNT, it was demonstrated that ELISA had a sensitivity and specificity of 100 and 92%, respectively. On the other hand, when the results obtained by our ELISA system were compared with those of an imported measles virus IgM ELISA kit (EIAgen; Adaltis Italia SPa, Bologna, Italy), a high level of agreement was shown (k = 0.926).

scholarly journals West Nile Virus and Related Flavivirus in European Wild Boar (Sus scrofa), Latium Region, Italy: A Retrospective Study

Animals ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 494
Author(s):  
Angela Petruccelli ◽  
Tiziana Zottola ◽  
Gianmarco Ferrara ◽  
Valentina Iovane ◽  
Cristina Di Russo ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Wild Boar ◽  
Specific Antibody ◽  
Central Italy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Serum Samples ◽  
West Nile ◽  
Wild Boars ◽  
European Wild Boar

Background: A retrospective sero-survey for evidence of West Nile virus (WNV) infection in European wild boar (Sus scorfa) was conducted in the Latium region, Italy, on stored serum samples of the period November 2011 to January 2012. Methods: Sera were collected from 168 European wild boars and screened for antibodies to WNV and other Flaviviruses by competitive enzyme linked immunosorbent assay (cELISA). All sera positive for Flavivirus antibodies by cELISA were further examined by virus neutralization test (VNT). To test the presence of Flavivirus RNA in samples, an RT-PCR was performed using a pan-Flavivirus primers pair. Results: Thirteen wild boars (7.73%) were seropositive for Flaviviruses. The hemolysis of serum samples limited the interpretation of the VNT for 7 samples, confirming the presence of specific antibody against WNV in a single European wild boar serum sample. The presence of ELISA positive/VNT negative samples suggests the occurrence of non-neutralizing antibodies against WNV or other antigen-related Flaviviruses. No samples resulted positive for Flavivirus by RT-PCR assay. Conclusion: Although a moderately high percentage of animals with specific antibody for WNV has been detected in wild boar in other surveillance studies in Europe, this has not been reported previously in Italy. Together, these data indicate that European wild boar are exposed to WNV and/or other related-Flavivirus in central Italy and confirm the usefulness of wild ungulates, as suitable Flavivirus sentinels.

scholarly journals Persistence of Antibodies to West Nile Virus in Naturally Infected Rock Pigeons (Columba livia)

2005 ◽  
Vol 12 (5) ◽  
pp. 665-667 ◽  
Author(s):  
Samantha E. J. Gibbs ◽  
Douglas M. Hoffman ◽  
Lillian M. Stark ◽  
Nicole L. Marlenee ◽  
Bradley J. Blitvich ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Immunosorbent Assay ◽  
Neutralization Test ◽  
Columba Livia ◽  
Maternal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
West Nile ◽  
Entire Study ◽  
Plaque Reduction Neutralization Test ◽  
Antibody Persistence

ABSTRACT Wild caught rock pigeons (Columba livia) with antibodies to West Nile virus were monitored for 15 months to determine antibody persistence and compare results of three serologic techniques. Antibodies persisted for the entire study as detected by epitope-blocking enzyme-linked immunosorbent assay and plaque reduction neutralization test. Maternal antibodies in squabs derived from seropositive birds persisted for an average of 27 days.

scholarly journals Induction of Epitope-Specific Neutralizing Antibodies against West Nile Virus

2007 ◽  
Vol 81 (21) ◽  
pp. 11828-11839 ◽  
Author(s):  
Theodore Oliphant ◽  
Grant E. Nybakken ◽  
S. Kyle Austin ◽  
Qing Xu ◽  
Jonathan Bramson ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Human Serum ◽  
Neutralizing Antibodies ◽  
Immunoglobulin M ◽  
Late Time ◽  
Loss Of Function ◽  
Serum Samples ◽  
West Nile ◽  
E Protein ◽  
Low Levels

ABSTRACT Previous studies have established that an epitope on the lateral ridge of domain III (DIII-lr) of West Nile virus (WNV) envelope (E) protein is recognized by strongly neutralizing type-specific antibodies. In contrast, an epitope against the fusion loop in domain II (DII-fl) is recognized by flavivirus cross-reactive antibodies with less neutralizing potential. Using gain- and loss-of-function E proteins and wild-type and variant WNV reporter virus particles, we evaluated the expression pattern and activity of antibodies against the DIII-lr and DII-fl epitopes in mouse and human serum after WNV infection. In mice, immunoglobulin M (IgM) antibodies to the DIII-lr epitope were detected at low levels at day 6 after infection. However, compared to IgG responses against other epitopes in DI and DII, which were readily detected at day 8, the development of IgG against DIII-lr epitope was delayed and did not appear consistently until day 15. This late time point is notable since almost all death after WNV infection in mice occurs by day 12. Nonetheless, at later time points, DIII-lr antibodies accumulated and comprised a significant fraction of the DIII-specific IgG response. In sera from infected humans, DIII-lr antibodies were detected at low levels and did not correlate with clinical outcome. In contrast, antibodies to the DII-fl were detected in all human serum samples and encompassed a significant percentage of the anti-E protein response. Our experiments suggest that the highly neutralizing DIII-lr IgG antibodies have little significant role in primary infection and that the antibody response of humans may be skewed toward the induction of cross-reactive, less-neutralizing antibodies.

scholarly journals West Nile Virus Seroprevalence among Qatari and Immigrant Populations within Qatar

2020 ◽  
Author(s):  
Hasna Kunhipurayil ◽  
Muna Ahmed ◽  
Gheyath Nasrallah
Keyword(s):  
West Nile Virus ◽  
Blood Donation ◽  
Enzyme Linked Immunosorbent Assay ◽  
Large Sample Size ◽  
Serum Samples ◽  
West Nile ◽  
Viral Neutralization ◽  
Healthy Donors ◽  
Confirmatory Tests ◽  
Sub Saharan

Background: West Nile virus (WNV) is one of the most widely spread arboviruses worldwide and a highly significant pathogen in humans and animals. Despite frequent outbreaks and endemic transmission being reported in the Middle East and North Africa (MENA), seroprevalence studies of WNV in Qatar are highly lacking. Aim: This study aims to investigate the actual prevalence of WNV among local and expatriate communities in the Qatar using a large sample size of seemingly healthy donors. Method: A total of 1992 serum samples were collected from donors of age 18 or older and were tested for the presence of WNV antibodies. Serion enzyme-linked immunosorbent assay (ELISA) commercial microplate kits were used to detect the presence of the WNV IgM and IgG. The seropositivity was statistically analyzed using SPSS software with a confidence interval of 95%. Results: The seroprevalence of anti-WNV IgG and IgM in Qatar was 10.3% and 3.4%, respectively. The country-specific seroprevalence according to nationality for WNV IgG and IgM, respectively, were Sudan (37.0%, 10.0%), Egypt (31.6%, 4.4%), India (13.4%, 3.2%), Yemen(10.2%, 7.0%), Pakistan (8.6%, 2.7%), Iran (10.6%, 0.0%), Philippines (5.4%, 0.0%), Jordan(6.8%, 1.1%), Syria (2.6%, 9.6%), Palestine (2.6%, 0.6%), Qatar (1.6%, 1.7%), and Lebanon (0.9%, 0.0%). The prevalence of both IgM and IgG was significantly correlated with the nationality (p≤0.001). Conclusion: Among these tested nationalities, Qatar national has a relatively low burden of WNV disease. The highest prevalence of WNV was found in the Sub Saharan African nationalities like Sudan and Egypt. The seroprevalence of WNV is different from the previously reported arboviruses such as CHIKV and DENV, which was highest among Asian countries (India and Philippines). Further confirmatory tests such as viral neutralization assays are needed to confirm the IgM seropositivity in these samples since these samples could be a source of viral transmission through blood donation.

scholarly journals Exposure to Zoonotic West Nile Virus in Long-Tailed Macaques and Bats in Peninsular Malaysia

Animals ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 2367
Author(s):  
Mohd Yuseri Ain-Najwa ◽  
Abd Rahaman Yasmin ◽  
Siti Suri Arshad ◽  
Abdul Rahman Omar ◽  
Jalila Abu ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Peninsular Malaysia ◽  
Wild Birds ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mangrove Forests ◽  
Rt Pcr ◽  
Serum Samples ◽  
West Nile ◽  
The Status

The role of wildlife such as wild birds, macaques, and bats in the spreading and maintenance of deadly zoonotic pathogens in nature have been well documented in many parts of the world. One such pathogen is the mosquitoes borne virus, namely the West Nile Virus (WNV). Previous research has shown that 1:7 and 1:6 Malaysian wild birds are WNV antibody and RNA positive, respectively, and bats in North America may not be susceptible to the WNV infection. This study was conducted to determine the status of WNV in Malaysian macaques and bats found in mangrove forests and caves, respectively. Archive sera and oropharyngeal swabs from long-tailed macaques were subjected to the antibody detection using WNV competitive enzyme-linked immunosorbent assay (c-ELISA) and WNV RNA using RT-PCR, respectively, while the archive oropharyngeal and rectal swabs from bats were subjected to RT-PCR without serological analysis due to the unavailability of serum samples. The analysis revealed a WNV seropositivity of 29.63% (24/81) and none of the macaques were positive for WNV RNA. Meanwhile, 12.2% (5/41) of the bats from Pteropodidae, Emballonuridae, and Rhinolophidae families tested positive for WNV RNA. Here, we show a high WNV antibody prevalence in macaques and a moderate WNV RNA in various Malaysian bat species, suggesting that WNV circulates through Malaysian wild animals and Malaysian bat species may be susceptible to the WNV infection.

scholarly journals Exposure of Horses in Israel to West Nile Virus and Usutu Virus

Viruses ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 1099
Author(s):  
Gili Schvartz ◽  
Sharon Tirosh-Levy ◽  
Oran Erster ◽  
Roni Shenhar ◽  
Hadas Levy ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Middle East ◽  
Neutralizing Antibodies ◽  
Mosquito Vectors ◽  
Serum Samples ◽  
West Nile ◽  
East Europe ◽  
Zoonotic Pathogen ◽  
Usutu Virus ◽  
Neuroinvasive Disease

West Nile virus (WNV) and Usutu virus (USUV) are arboviruses transmitted by mosquito vectors. Whereas WNV is endemic in Israel, the Middle East, Europe, and in the Americas, data regarding the prevalence of USUV in the Middle East is limited. While both viruses share similar reservoirs and vectors, exposure of horses in the area to USUV have never been assessed. The aim of this study was to estimate the seroprevalence and co-exposure of WNV and USUV in horses in Israel. A total of 327 serum samples from healthy unvaccinated horses in Israel collected in 2018 were tested for neutralizing antibodies against WNV and USUV. Seroprevalence for neutralizing antibodies against WNV and USUV was 84.1% and 10.8%, respectively. Management and age were significantly associated with WNV and USUV seropositivity. This is the first report describing exposure of horses in Israel to USUV, which indicates that this zoonotic pathogen should be included in the differential diagnosis list of neuroinvasive disease in this country.

Sign in / Sign up

Export Citation Format

Share Document