scholarly journals Duplex Microsphere-Based Immunoassay for Detection of Anti-West Nile Virus and Anti-St. Louis Encephalitis Virus Immunoglobulin M Antibodies

2005 ◽  
Vol 12 (5) ◽  
pp. 566-574 ◽  
Author(s):  
Alison J. Johnson ◽  
Amanda J. Noga ◽  
Olga Kosoy ◽  
Robert S. Lanciotti ◽  
Alicia A. Johnson ◽  
...  
Keyword(s):  
New York ◽  
Neutralization Test ◽  
Rapid Test ◽  
The United States ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
West Nile ◽  
Plaque Reduction Neutralization Test ◽  
Viral Antigens ◽  
Primary Test

ABSTRACT West Nile (WN) virus was introduced into the United States in 1999, when the first human cases of WN fever and encephalitis appeared in New York City. From there, the virus has spread throughout North America, in some areas cocirculating with the related flavivirus St. Louis encephalitis (SLE) virus. Public health laboratories currently use an immunoglobulin M (IgM) antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) as a primary test for human serodiagnosis, followed by a confirmatory plaque-reduction neutralization test (PRNT). The MAC-ELISAs take 2 days to perform; therefore there is a need for a more rapid test. This report describes a duplex microsphere-based immunoassay (MIA) that shortens the test processing time to about 4.5 h. The assay employs two sets of microspheres coupled to a single flavivirus group-reactive antibody, which are used to capture the WN and SLE viral antigens independently. Immunoglobulin G-depleted serum is concurrently assayed for IgM antibodies to each of the viral antigens. The results are standardized and classified by using quadratic discriminant analysis so that a single result, anti-WN IgM-positive, anti-SLE IgM-positive, negative, or nonspecific, can be determined. The duplex MIA results compared favorably to those of the plaque-reduction neutralization test and MAC-ELISA. The assay proved to be reproducible, produced accurate classifications as to the infecting virus, and was specific.

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 Performance Characteristics of an In-House Assay System Used To Detect West Nile Virus (WNV)-Specific Immunoglobulin M during the 2001 WNV Season in the United States

2003 ◽  
Vol 10 (1) ◽  
pp. 177-179 ◽  
Author(s):  
Harry E. Prince ◽  
Wayne R. Hogrefe
Keyword(s):  
West Nile Virus ◽  
The United States ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Screening Assay ◽  
West Nile ◽  
Elisa System ◽  
Igm Elisa ◽  
Specific Immunoglobulin ◽  
Control And Prevention

ABSTRACT During the 2001 U. S. West Nile virus (WNV) season, 163 specimens were reactive in an in-house WNV-specific immunoglobulin M (IgM) screening enzyme-linked immunosorbent assay (ELISA) and were referred to either the Centers for Disease Control and Prevention or the appropriate state public health laboratory (CDC/SPHL) for additional testing. CDC/SPHL supplied results for 124 specimens that could be further evaluated in-house: 70 specimens were nonreactive in the CDC/SPHL WNV-specific IgM screening assay, and 54 specimens were reactive. These specimens were used to evaluate a modified in-house WNV-specific IgM ELISA that incorporated background subtraction to identify nonspecific reactivity and thus improve assay specificity. Of the 70 CDC/SPHL nonreactive samples, 49 (70%) were nonreactive in the modified ELISA; of the 54 CDC/SPHL reactive samples, 51 (94%) were reactive in the modified ELISA. Confirmatory studies performed by CDC/SPHL indicated that 38 CDC/SPHL screen-reactive specimens represented true WNV infection; all 38 specimens were reactive in the modified in-house WNV-specific IgM ELISA. These findings demonstrate that an in-house ELISA system for WNV-specific IgM effectively identifies patients with WNV infection.

scholarly journals Immunoglobulin G Avidity in Differentiation between Early and Late Antibody Responses to West Nile Virus

2006 ◽  
Vol 13 (1) ◽  
pp. 33-36 ◽  
Author(s):  
Janet L. Fox ◽  
Stuart L. Hazell ◽  
Leslie H. Tobler ◽  
Michael P. Busch
Keyword(s):  
United States ◽  
New York ◽  
West Nile Virus ◽  
The United States ◽  
Immunoglobulin M ◽  
West Nile ◽  
Marked Rise ◽  
Igm Antibodies ◽  
Igg Avidity ◽  
Igg Level

ABSTRACT In 1999 West Nile virus (WNV) surfaced in the United States in the city of New York and spread over successive summers to most of the continental United States, Canada, and Mexico. Because WNV immunoglobulin M (IgM) antibodies have been shown to persist for up to 1 year, residents in areas of endemicity can have persistent WNV IgM antibodies that are unrelated to a current illness with which they present. We present data on the use of IgG avidity testing for the resolution of conflicting data arising from the testing of serum or plasma for antibodies to WNV. Thirteen seroconversion panels, each consisting of a minimum of four samples, were used. All samples were tested for the presence of WNV IgM and IgG antibodies, and the avidity index for the WNV IgG-positive samples was calculated. Panels that exhibited a rise in the WNV IgM level followed by a sequential rise in the WNV IgG level were designated “primary.” Panels that exhibited a marked rise in the WNV IgG level followed by a sequential weak WNV IgM response and that had serological evidence of a prior flavivirus infection were designated “secondary.” All samples from the “primary” panels exhibited low avidity indices (less than 40%) for the first 20 to 30 days after the recovery of the index sample (the sample found to be virus positive). All of the “secondary” samples had elevated WNV IgG levels with avidity indices of ≥55%, regardless of the number of days since the recovery of the index sample. These data demonstrate that it is possible to differentiate between recent and past exposure to WNV or another flavivirus through the measurement of WNV IgG avidity indices.

scholarly journals Evaluation of a Diagnostic Algorithm Using Immunoglobulin M Enzyme-Linked Immunosorbent Assay To Differentiate Human West Nile Virus and St. Louis Encephalitis Virus Infections during the 2002 West Nile Virus Epidemic in the United States

2004 ◽  
Vol 11 (6) ◽  
pp. 1130-1133 ◽  
Author(s):  
Denise A. Martin ◽  
Amanda Noga ◽  
Olga Kosoy ◽  
Alison J. Johnson ◽  
Lyle R. Petersen ◽  
...  
Keyword(s):  
United States ◽  
West Nile Virus ◽  
Immunosorbent Assay ◽  
Diagnostic Algorithm ◽  
The United States ◽  
Encephalitis Virus ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Infections ◽  
West Nile

ABSTRACT A diagnostic algorithm was developed to differentiate between human infections of West Nile virus (WNV) and St. Louis encephalitis virus (SLEV) using positive-to-negative (P/N) ratios derived from the immunoglobulin M capture enzyme-linked immunosorbent assay (MAC-ELISA). To validate this algorithm, we tested 1,418 serum and cerebrospinal fluid (CSF) samples from confirmed WNV and SLEV infections collected during the WNV epidemic of 2002 in the United States. WNV P/N-to-SLEV P/N ratios (W/S ratios) were calculated and used to identify the infecting virus. These results were compared to results from the plaque reduction neutralization test (PRNT), which is currently the standard assay used to discriminate between closely related flavivirus infections. If the W/S ratio was ≥1, the predictive value positive (PNP) for WNV was 97.8%, where 95% of flavivirus cases were due to WNV infection and only 3.7% of specimens would require PRNT to differentiate WNV from SLEV infection. Use of the W/S ratio as part of the testing algorithm to interpret MAC-ELISA results generates reportable probable cases quickly, alleviating the need for PRNT in most instances.

scholarly journals Detection of West Nile Virus (WNV)-Specific Immunoglobulin M in a Reference Laboratory Setting during the 2002 WNV Season in the United States

2003 ◽  
Vol 10 (5) ◽  
pp. 764-768 ◽  
Author(s):  
Harry E. Prince ◽  
Wayne R. Hogrefe
Keyword(s):  
West Nile Virus ◽  
Serum Sample ◽  
The United States ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Reference Laboratory ◽  
Sample Collection ◽  
Serum Samples ◽  
West Nile ◽  
Specific Immunoglobulin

ABSTRACT Between 1 June and 31 December 2002, 30,677 serum samples and 4,554 cerebrospinal fluid (CSF) samples were tested for West Nile virus (WNV)-specific immunoglobulin M (IgM) by an in-house enzyme-linked immunosorbent assay (ELISA); 1,481 serum samples (4.8%) and 345 CSF samples (7.6%) were positive for WNV IgM. Positive samples were forwarded to public health service laboratories (PHSLs) for further testing. PHSLs supplied results from their WNV IgM ELISAs for 654 samples; 633 (97%) were positive. PHSLs supplied WNV plaque reduction neutralization test results for 128 samples; 123 (96%) were positive. WNV IgM seroconversion and seroreversion trends were evaluated for 749 patients who each provided two serum samples that were tested during the study period. Of 574 patients whose first serum sample was IgM negative, 41 (7%) seroconverted (the second serum sample was IgM positive); of 175 patients whose first serum sample was IgM positive, 22 (13%) seroreverted (the second serum sample was IgM negative). The seroreversion rate was directly proportional to the time between serum sample collection; whereas only 1% of patients whose sera were collected <20 days apart showed seroreversion, 54% of patients whose sera were collected >60 days apart showed seroreversion. Conversion and reversion trends for CSF were evaluated for 68 patients. Of 54 patients whose first CSF specimen was IgM negative, 9 (17%) converted; none of 14 patients whose first CSF specimen was IgM positive reverted. Concomitant detection of WNV IgM in serum and CSF was assessed for 1,188 patients for whom paired serum and CSF specimens were available; for all 130 patients for whom IgM was detectable in CSF, IgM was also detectable in serum. These findings show that an in-house WNV IgM ELISA accurately identifies patients with WNV infection, document WNV IgM conversion and reversion trends, and demonstrate that WNV IgM detection in CSF is accompanied by WNV IgM detection in serum.

scholarly journals First Report of Grapevine leafroll associated virus-3 in American Vitis spp. Grapevines in Washington State

Plant Disease ◽  
2006 ◽  
Vol 90 (11) ◽  
pp. 1461-1461 ◽  
Author(s):  
M. J. Soule ◽  
K. C. Eastwell ◽  
R. A. Naidu
Keyword(s):  
New York ◽  
Economic Impact ◽  
Virus Disease ◽  
The United States ◽  
Washington State ◽  
The State ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
First Report ◽  
Table Grapes

Washington State is the largest producer of juice grapes (Vitis labruscana ‘Concord’ and Vitis labrusca ‘Niagara’) and ranks second in wine grape production in the United States. Grapevine leafroll disease (GLD) is the most wide spread and economically significant virus disease in wine grapes in the state. Previous studies (2) have shown that Grapevine leafroll associated virus-3 (GLRaV-3) is the predominant virus associated with GLD. However, little is known about the incidence and economic impact of GLD on juice and table grapes. Because typical GLD symptoms may not be obvious among these cultivars, the prevalence and economic impact of GLD in Concord and Niagara, the most widely planted cultivars in Washington State, has received little attention from the grape and nursery industries. During the 2005 growing season, 32 samples from three vineyards and one nursery of ‘Concord’ and three samples from one nursery of ‘Niagara’ were collected randomly. Petiole extracts were tested by single-tube reverse transcription-polymerase chain reaction (RT-PCR; 3) with primers LC 1 (5′-CGC TAG GGC TGT GGA AGT ATT-3′) and LC 2 (5′-GTT GTC CCG GGT ACC AGA TAT-3′), specific for the heat shock protein 70 homologue (Hsp70h gene) of GLRaV-3 (GenBank Accession No. AF037268). One ‘Niagara’ nursery sample and eleven ‘Concord’ samples from the three vineyards tested positive for GLRaV-3, producing a single band of the expected size of 546 bp. The ‘Niagara’ and six of the ‘Concord’ RT-PCR products were cloned in pCR2.1 (Invitrogen Corp, Carlsbad, CA) and the sequences (GenBank Accession Nos. DQ780885, DQ780886, DQ780887, DQ780888, DQ780889, DQ780890, and DQ780891) compared with the respective sequence of a New York isolate of GLRaV-3 (GenBank Accession No. AF037268). The analysis revealed that GLRaV-3 isolates from ‘Concord’ and ‘Niagara’ share nucleotide identities of 94 to 98% and amino acid identities and similarities of 97 to 98% with the Hsp70h gene homologue of the New York isolate of GLRaV-3. Additional testing by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) using antibodies specific to GLRaV-3 (BIOREBA AG, Reinach, Switzerland) further confirmed these results in the ‘Niagara’ and two of the ‘Concord’ isolates. GLRaV-3 has previously been reported in labrusca cvs. Concord and Niagara in western New York (4) and Canada (1), but to our knowledge, this is the first report of GLRaV-3 in American grapevine species in the Pacific Northwest. Because wine and juice grapes are widely grown in proximity to each other in Washington State and grape mealybug (Pseudococcus maritimus), the putative vector of GLRaV-3, is present in the state vineyards, further studies will focus on the role of American grapevine species in the epidemiology of GLD. References: (1) D. J. MacKenzie et al. Plant Dis. 80:955, 1996. (2) R. R. Martin et al. Plant Dis. 89:763, 2005. (3) A. Rowhani et al. ICGV, Extended Abstracts, 13:148, 2000. (4) W. F. Wilcox et al. Plant Dis. 82:1062, 1998.

scholarly journals Potential Antigenic Cross-reactivity Between Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) and Dengue Viruses

2020 ◽  
Author(s):  
Yaniv Lustig ◽  
Shlomit Keler ◽  
Rachel Kolodny ◽  
Nir Ben-Tal ◽  
Danit Atias-Varon ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Envelope Protein ◽  
False Positive ◽  
In Silico ◽  
Rapid Test ◽  
Cross Reactivity ◽  
Lateral Flow ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Testing

Abstract Background Coronavirus disease 2019 (COVID-19) and dengue fever are difficult to distinguish given shared clinical and laboratory features. Failing to consider COVID-19 due to false-positive dengue serology can have serious implications. We aimed to assess this possible cross-reactivity. Methods We analyzed clinical data and serum samples from 55 individuals with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. To assess dengue serology status, we used dengue-specific antibodies by means of lateral-flow rapid test, as well as enzyme-linked immunosorbent assay (ELISA). Additionally, we tested SARS-CoV-2 serology status in patients with dengue and performed in-silico protein structural analysis to identify epitope similarities. Results Using the dengue lateral-flow rapid test we detected 12 positive cases out of the 55 (21.8%) COVID-19 patients versus zero positive cases in a control group of 70 healthy individuals (P = 2.5E−5). This includes 9 cases of positive immunoglobulin M (IgM), 2 cases of positive immunoglobulin G (IgG), and 1 case of positive IgM as well as IgG antibodies. ELISA testing for dengue was positive in 2 additional subjects using envelope protein directed antibodies. Out of 95 samples obtained from patients diagnosed with dengue before September 2019, SARS-CoV-2 serology targeting the S protein was positive/equivocal in 21 (22%) (16 IgA, 5 IgG) versus 4 positives/equivocal in 102 controls (4%) (P = 1.6E−4). Subsequent in-silico analysis revealed possible similarities between SARS-CoV-2 epitopes in the HR2 domain of the spike protein and the dengue envelope protein. Conclusions Our findings support possible cross-reactivity between dengue virus and SARS-CoV-2, which can lead to false-positive dengue serology among COVID-19 patients and vice versa. This can have serious consequences for both patient care and public health.

A Single-Center Experience with a Pregnant Immigrant Population and Zika Virus Serologic Screening in New York City

2019 ◽  
Vol 37 (07) ◽  
pp. 731-737 ◽  
Author(s):  
Audrey A. Merriam ◽  
Chia-Ling Nhan-Chang ◽  
B. Isabel Huerta-Bogdan ◽  
Ronald Wapner ◽  
Cynthia Gyamfi-Bannerman
Keyword(s):  
At Risk ◽  
New York ◽  
New York City ◽  
York City ◽  
Zika Virus ◽  
Immunoglobulin M ◽  
Immigrant Population ◽  
Enzyme Linked Immunosorbent Assay ◽  
Neurodevelopmental Outcomes ◽  
Positive Urine

Objective Our institution is in an area of New York City with a large population of immigrants from Zika virus endemic areas. With the recent Zika virus outbreak, we sought to examine our center's experience with screening for Zika virus and outcomes among patients who tested positive for the disease during pregnancy. Study Design We performed a chart review of all pregnant patients who tested positive (positive serum or urine polymerase chain reaction [PCR]) or presumed positive (immunoglobulin M [IgM] enzyme-linked immunosorbent assay [ELISA] positive or IgM ELISA equivocal with positive plaque reduction neutralization test) for Zika virus. All tests were performed by the Department of Health (DOH) and followed Centers for Disease Control and Prevention guidelines in effect at the time of specimen collection. Testing of cord blood, placenta, and/or neonatal blood were/was performed by the DOH for New York County. Prenatal ultrasounds for fetal head size and surveillance for calcifications were performed by maternal–fetal medicine specialists. Infant head ultrasound results were included when available. Results Between March 2016 and April 2017, 70 pregnant patients were positive or presumed positive for Zika infection during pregnancy. Of those, 16 women had positive urine or serum PCR and the remaining 54 were presumed positive. Among positive cases, five women tested positive via urine PCR only, nine women tested positive via serum PCR only, and two women had both positive urine and serum PCR. Fifteen of 67 infants (22%) born during the study period were born to mothers with positive urine or serum PCR testing. Sixty-five newborns were clinically normal with normal head measurements. Of the intracranial ultrasound performed, one infant had a grade 1 intraventricular hemorrhage, four had incidental choroid plexus cysts, and one had severe ventriculomegaly that was also noted antenatally. There were 2 positive and 15 equivocal infant serum IgM samples and 1 positive placental PCR from these pregnancies. There were four pregnancy terminations and two cases with fetal anomalies in this population that were split evenly between patients who tested positive and those who tested presumed positive for Zika virus during pregnancy. Conclusion We found no differences in pregnancy or neonatal outcomes between women who tested positive and presumed positive for Zika virus during pregnancy. Testing of infants and placenta tissue after delivery was largely inconclusive. Improvement in testing for Zika virus infection is needed to determine which pregnancies are at risk for congenital anomalies. Further research is still needed to determine which children are at risk for poor neurodevelopmental outcomes related to Zika virus and how to best coordinate care among the immigrant population during a new disease epidemic.

scholarly journals West Nile Virus Recombinant DNA Vaccine Protects Mouse and Horse from Virus Challenge and Expresses In Vitro a Noninfectious Recombinant Antigen That Can Be Used in Enzyme-Linked Immunosorbent Assays

2001 ◽  
Vol 75 (9) ◽  
pp. 4040-4047 ◽  
Author(s):  
Brent S. Davis ◽  
Gwong-Jen J. Chang ◽  
Bruce Cropp ◽  
John T. Roehrig ◽  
Denise A. Martin ◽  
...  
Keyword(s):  
United States ◽  
Recombinant Plasmid ◽  
Animal Health ◽  
The United States ◽  
Virus Antigen ◽  
Recombinant Antigen ◽  
Immunoglobulin M ◽  
E Proteins ◽  
West Nile ◽  
Immunosorbent Assays

ABSTRACT Introduction of West Nile (WN) virus into the United States in 1999 created major human and animal health concerns. Currently, no human or veterinary vaccine is available to prevent WN viral infection, and mosquito control is the only practical strategy to combat the spread of disease. Starting with a previously designed eukaryotic expression vector, we constructed a recombinant plasmid (pCBWN) that expressed the WN virus prM and E proteins. A single intramuscular injection of pCBWN DNA induced protective immunity, preventing WN virus infection in mice and horses. Recombinant plasmid-transformed COS-1 cells expressed and secreted high levels of WN virus prM and E proteins into the culture medium. The medium was treated with polyethylene glycol to concentrate proteins. The resultant, containing high-titered recombinant WN virus antigen, proved to be an excellent alternative to the more traditional suckling-mouse brain WN virus antigen used in the immunoglobulin M (IgM) antibody-capture and indirect IgG enzyme-linked immunosorbent assays. This recombinant antigen has great potential to become the antigen of choice and will facilitate the standardization of reagents and implementation of WN virus surveillance in the United States and elsewhere.

Sign in / Sign up

Export Citation Format

Share Document