Sensitivity and Kinetics of an NS1-Based Zika Virus Enzyme-Linked Immunosorbent Assay in Zika Virus-Infected Travelers from Israel, the Czech Republic, Italy, Belgium, Germany, and Chile

ABSTRACT Serological diagnosis of Zika virus is challenging due to high cross-reactivity of Zika virus with other flavivirus antibodies. Recently, a Zika NS1-based enzyme-linked immunosorbent assay (ELISA) was developed and shown to be highly specific for Zika antibody detection; however, sensitivity was evaluated for only a small number of confirmed Zika-infected patients. In this study, we measured the sensitivity and kinetics of Zika IgM and IgG antibodies using the Zika NS1-based ELISA in 105 samples from 63 returning travelers infected with Zika virus (proven by PCR or neutralization assay) from Israel, Czech Republic, Italy, Belgium, Germany, and Chile. Zika virus IgM was detected from 2 to 42 days post-symptom onset (PSO) with an overall sensitivity of 79% in the first month and 68% until 2 months PSO, while IgG antibodies were detected from 5 days to 3 years PSO with 79% sensitivity. Interestingly, significant differences in IgM sensitivity and IgM detection period were observed between Israeli and European/Chilean Zika-infected travelers, adding to the complexity of Zika infection diagnosis and suggesting that other diagnostic methods should be complemented to reduce false-negative results.

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Detection and Titration of Human Herpesvirus-8–Specific Antibodies in Sera From Blood Donors, Acquired Immunodeficiency Syndrome Patients, and Kaposi's Sarcoma Patients Using a Whole Virus Enzyme-Linked Immunosorbent Assay

Blood ◽

10.1182/blood.v92.1.53.413k30_53_58 ◽

1998 ◽

Vol 92 (1) ◽

pp. 53-58 ◽

Cited By ~ 98

Author(s):

Louise G. Chatlynne ◽

William Lapps ◽

Michael Handy ◽

Yao Q. Huang ◽

Rizwan Masood ◽

...

Keyword(s):

Immunosorbent Assay ◽

Kaposi's Sarcoma ◽

Blood Donors ◽

Human Herpesvirus ◽

Kaposi’S Sarcoma ◽

Antibody Detection ◽

Human Herpesvirus 8 ◽

Enzyme Linked Immunosorbent Assay ◽

Igg Antibodies ◽

Herpesvirus 8

A human herpesvirus-8 (HHV-8) enzyme-linked immunosorbent assay (ELISA) with a whole virus lysate as antigen was developed and used to measure the seroprevalence rate and levels of IgG antibodies to HHV-8 in sera/plasma of various patient groups and blood donors. The virus antigen was prepared from the KS-1 cell line, which produces lytic virus, and therefore contains a broad array of viral proteins. Seroprevalence studies using this ELISA showed the following: 10 of 91 blood donors (11%) had an average HHV-8 antibody titer of 118; 67 of 72 (93%) classic Kaposi's sarcoma (KS) patients were positive with an average titer of 14,111; and 57 of 62 (92%) KS/human immunodeficiency virus (HIV) patients were positive with an average titer of 4,000. A study on a very limited number of serial serum samples from patients before and after diagnosis with KS showed highly elevated antibody titers to HHV-8 virus after KS lesions developed. Preliminary data show that 50% of the sera from HIV-1+ homosexual patients contain IgG antibodies to HHV-8 suggesting that this population is at high risk for developing KS. Antibody results correlated well with the confirmatory immunofluorescent assays (IFA) using KS-1 cells as the substrate. This HHV-8 IgG antibody detection ELISA is sensitive and specific and does not cross-react with Epstein-Barr virus (EBV) or other human herpesviruses. The results of this HHV-8 antibody survey suggest that this rapid ELISA assay can be used to screen large numbers of sera to find those at risk for developing KS.

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An Enzyme-Linked Immunosorbent Assay for the Detection of Aspergillus parasiticus and Aspergillus flavus

Journal of Food Protection ◽

10.4315/0362-028x-60.8.978 ◽

1997 ◽

Vol 60 (8) ◽

pp. 978-984 ◽

Cited By ~ 15

Author(s):

GUO-JANE TSAI ◽

SHOU-CHIN YU

Keyword(s):

Aspergillus Flavus ◽

Immunosorbent Assay ◽

False Positive ◽

Aspergillus Parasiticus ◽

False Negative ◽

Zealand White Rabbit ◽

Cross Reactivity ◽

Enzyme Linked Immunosorbent Assay ◽

Molecular Weights ◽

Sds Page

An enzyme-linked immunosorbent assay (ELISA) was established for the specific detection of Aspergillus parasiticus and Aspergillus flavus. A New Zealand white rabbit was immunized intravenously with 100 μg of A. parasiticus CCRC 30117 mycelial protein extracts. The antibodies were separated and purified. The optimal concentration of the antibody and antibody-peroxidase conjugate used in the established ELISA was 10 μg/ml with a detection limit of 1 μg/ml. Among the 126 strains tested (including 21 strains of A. parasiticus, 11 strains of A. flavus, 34 isolates of A. parasiticus/A. flavus from cereals, and 60 strains of non-A. parasiticus/A. flavus fungi), the false-negative and false-positive rates were 1.5 and 3.3%, respectively. Strains of Aspergillus flavofrucatis and Aspergillus sojae produced false-positive reactions. However, their antigens had much lower cross-reactivity with the antibodies raised against A. parasiticus, as shown from I50 values. The molecular weights of the main antigens of A. parasiticus were 94, 82, and 40 kDa. The two heavier antigens had higher sugar contents, as demonstrated by SDS-PAGE and immunoblotting. A good correlation (r = 0.97) was found between mycelium measurement by weighing and by ELISA for A. parasiticus grown in yeast extract sucrose broth (YESB) at 25°C.

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First detection of Zika virus infection in a Croatian traveler returning from Brazil, 2016

The Journal of Infection in Developing Countries ◽

10.3855/jidc.9410 ◽

2017 ◽

Vol 11 (08) ◽

pp. 662-667

Author(s):

Tatjana Vilibic-Cavlek ◽

Ljiljana Betica-Radic ◽

Giulietta Venturi ◽

Claudia Fortuna ◽

Stjepan Djuricic ◽

...

Keyword(s):

Zika Virus ◽

High Titer ◽

Disease Onset ◽

Negative Control ◽

Cross Reactivity ◽

Immunofluorescence Assay ◽

Enzyme Linked Immunosorbent Assay ◽

Low Grade ◽

Igg Antibodies ◽

Zikv Infection

In the last few years, several imported cases of Zika virus (ZIKV) infection were reported in European countries. We report the first imported ZIKV infection case in a Croatian traveler returning from Brazil. The patient presented with a low-grade fever, pruritic rash, general weakness, myalgia, arthralgia and edema of the legs and recovered completely within a week. ZIKV infection was confirmed by detection of IgM/IgG antibodies using enzyme-linked immunosorbent assay (ELISA) and confirmed by plaque-reduction neutralization test (PRNT). ZIKV IgM antibodies cross-reacted with dengue virus (DENV), West Nile virus (WNV) and tick-borne encephalitis virus (TBEV) in ELISA. In indirect immunofluorescence assay (IFA), IgM cross-reactivity was found only with DENV-3. ZIKV IgG antibodies cross-reacted with DENV in both ELISA and IFA. PRNT for DENV was negative. Control serology performed on days 64 and 98 after disease onset showed a decline in cross-reactive heterologous DENV IgG antibodies compared to persistently high titer of homologous ZIKV IgG antibodies.

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Diagnosis of Zika Virus Infection by Peptide Array and Enzyme-Linked Immunosorbent Assay

mBio ◽

10.1128/mbio.00095-18 ◽

2018 ◽

Vol 9 (2) ◽

Cited By ~ 33

Author(s):

Nischay Mishra ◽

Adrian Caciula ◽

Adam Price ◽

Riddhi Thakkar ◽

James Ng ◽

...

Keyword(s):

Yellow Fever ◽

Immunosorbent Assay ◽

Zika Virus ◽

Natural Infection ◽

Cross Reactivity ◽

High Density ◽

Serological Diagnosis ◽

Enzyme Linked Immunosorbent Assay ◽

Molecular Testing ◽

West Nile

ABSTRACTZika virus (ZIKV) is implicated in fetal stillbirth, microcephaly, intracranial calcifications, and ocular anomalies following vertical transmission from infected mothers. In adults, infection may trigger autoimmune inflammatory polyneuropathy. Transmission most commonly follows the bite of infectedAedesmosquitoes but may also occur through sexual intercourse or receipt of blood products. Definitive diagnosis through detection of viral RNA is possible in serum or plasma within 10 days of disease onset, in whole blood within 3 weeks of onset, and in semen for up to 3 months. Serological diagnosis is nonetheless critical because few patients have access to molecular diagnostics during the acute phase of infection and infection may be associated with only mild or inapparent disease that does not prompt molecular testing. Serological diagnosis is confounded by cross-reactivity of immune sera with other flaviviruses endemic in the areas where ZIKV has recently emerged. Accordingly, we built a high-density microarray comprising nonredundant 12-mer peptides that tile, with one-residue overlap, the proteomes of Zika, dengue, yellow fever, West Nile, Ilheus, Oropouche, and chikungunya viruses. Serological analysis enabled discovery of a ZIKV NS2B 20-residue peptide that had high sensitivity (96.0%) and specificity (95.9%) versus natural infection with or vaccination against dengue, chikungunya, yellow fever, West Nile, tick-borne encephalitis, or Japanese encephalitis virus in a microarray assay and an enzyme-linked immunosorbent assay (ELISA) of early-convalescent-phase sera (2 to 3 weeks after onset of symptomatic infection).IMPORTANCEThe emergence of Zika virus (ZIKV) as a teratogen is a profound challenge to global public health. Molecular diagnosis of infection is straightforward during the 3-week period when patients are viremic. However, serological diagnosis thereafter of historical exposure has been confounded by cross-reactivity. Using high-density peptide arrays that tile the proteomes of a selection of flaviviruses to identify a ZIKV-specific peptide, we established two assays that enable sensitive and specific diagnosis of exposure to ZIKV. These assays may be useful in guiding clinical management of mothers at risk for potential exposure to ZIKV and enable insights into the epidemiology of ZIKV infections.

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Enzyme-Linked Immunosorbent Assay for Detection of Filovirus Species-Specific Antibodies

Clinical and Vaccine Immunology ◽

10.1128/cvi.00170-10 ◽

2010 ◽

Vol 17 (11) ◽

pp. 1723-1728 ◽

Cited By ~ 72

Author(s):

Eri Nakayama ◽

Ayaka Yokoyama ◽

Hiroko Miyamoto ◽

Manabu Igarashi ◽

Noriko Kishida ◽

...

Keyword(s):

Ebola Virus ◽

Cross Reactivity ◽

Marburg Virus ◽

Diagnostic Methods ◽

Enzyme Linked Immunosorbent Assay ◽

Virus Species ◽

Igg Antibodies ◽

Specific Antibodies ◽

Species Specific ◽

Respective Species

ABSTRACT Several enzyme-linked immunosorbent assays (ELISAs) for the detection of filovirus-specific antibodies have been developed. However, diagnostic methods to distinguish antibodies specific to the respective species of filoviruses, which provide the basis for serological classification, are not readily available. We established an ELISA using His-tagged secreted forms of the transmembrane glycoproteins (GPs) of five different Ebola virus (EBOV) species and one Marburg virus (MARV) strain as antigens for the detection of filovirus species-specific antibodies. The GP-based ELISA was evaluated by testing antisera collected from mice immunized with virus-like particles as well as from humans and nonhuman primates infected with EBOV or MARV. In our ELISA, little cross-reactivity of IgG antibodies was observed in most of the mouse antisera. Although sera and plasma from some patients and monkeys showed notable cross-reactivity with the GPs from multiple filovirus species, the highest reactions of IgG were uniformly detected against the GP antigen homologous to the virus species that infected individuals. We further confirmed that MARV-specific IgM antibodies were specifically detected in specimens collected from patients during the acute phase of infection. These results demonstrate the usefulness of our ELISA for diagnostics as well as ecological and serosurvey studies.

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Development of an Enzyme-Linked Immunosorbent Assay To Detect Antibodies Targeting Recombinant Envelope Protein 2 of Mayaro Virus

Journal of Clinical Microbiology ◽

10.1128/jcm.01892-18 ◽

2019 ◽

Vol 57 (5) ◽

Cited By ~ 8

Author(s):

Marcílio Jorge Fumagalli ◽

William Marciel de Souza ◽

Marília Farignoli Romeiro ◽

Michell Charles de Souza Costa ◽

Renata Dezengrini Slhessarenko ◽

...

Keyword(s):

Immunosorbent Assay ◽

Envelope Protein ◽

Cross Reactivity ◽

Antibody Detection ◽

Enzyme Linked Immunosorbent Assay ◽

High Avidity ◽

Serum Samples ◽

Igg Antibody ◽

Mayaro Virus ◽

Recombinant Envelope Protein

ABSTRACT Mayaro virus (MAYV) is a neglected arthropod-borne virus (arbovirus) antigenically clustered into the Semliki Forest complex group of Alphavirus genus (Togaviridae family), maintained in an unclear zoonotic cycle involving mosquitoes from Haemagogus genus as the main vector. The genome is composed of a positive single-stranded RNA of 11.5 kb in length, which contains two genes that encode four nonstructural (nsP1 to nsP4) and five structural (C, E3, E2, 6K, and E1) proteins. In the present study, we have developed an enzyme-linked immunosorbent assay (ELISA) using as antigen the recombinant envelope protein 2 of MAYV produced in an Escherichia coli system (rE2-MAYV ELISAs). A panel of 68 human serum samples from suspected arboviral cases was analyzed and titrated for anti-MAYV IgM and IgG antibody detection. The rE2-MAYV ELISA detected 33.8% (23/68) IgG-positive samples, demonstrating 100% sensitivity and 78.95% specificity compared to the MAYV-specific 50% plaque reduction neutralization assay. In addition, the positive MAYV-neutralizing samples showed high titers of detection by rE2-MAYV ELISA, suggesting a highly sensitive test. The rE2-MAYV ELISA also detected 42.5% (29/68) IgM-positive samples, of which 13.8% (4/29) presented high-avidity interactions with rE2-MAYV. Cross-reactivity was observed with Chikungunya virus (CHIKV)-specific murine antibody sample but not with CHIKV-specific human and other Alphavirus murine antibodies. In short, we have developed a rapid, simple, specific, and sensitive MAYV rE2-ELISA, and our preliminary results show its potential applicability to diagnosis of MAYV infections.

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Accuracy of the Zika IgM Antibody Capture Enzyme-Linked Immunosorbent Assay from the Centers for Disease Control and Prevention (CDC Zika MAC-ELISA) for Diagnosis of Zika Virus Infection

Diagnostics ◽

10.3390/diagnostics10100835 ◽

2020 ◽

Vol 10 (10) ◽

pp. 835

Author(s):

Moyra Machado Portilho ◽

Laise de Moraes ◽

Mariana Kikuti ◽

Leile Camila Jacob Nascimento ◽

Mitermayer Galvão Reis ◽

...

Keyword(s):

Immunosorbent Assay ◽

Zika Virus ◽

Cross Reactivity ◽

Serological Diagnosis ◽

Enzyme Linked Immunosorbent Assay ◽

Rt Pcr ◽

Zikv Infection ◽

Igm Antibody ◽

Convalescent Phase ◽

Antibody Capture

Serological diagnosis of Zika virus (ZIKV) infection is challenging because of antigenic cross-reactivity with dengue virus (DENV). This study evaluated the accuracy of the Zika IgM antibody capture enzyme-linked immunosorbent assay (CDC Zika IgM MAC-ELISA) in differentiating between ZIKV and DENV infections. To determine sensitivity, we used acute- and convalescent-phase sera from 21 patients with RT-PCR-confirmed ZIKV infection. To determine specificity, we used acute- and convalescent-phase sera from 60 RT-PCR-confirmed dengue cases and sera from 23 blood donors. During the acute-phase of the illness, the assay presented a sensitivity of 12.5% (2/16) for samples collected 0–4 days post symptoms onset (DPSO), and of 75.0% (3/4) for samples collected 5–9 DPSO. During the convalescent-phase of the illness, the test sensitivity was 90.9% (10/11), 100% (2/2), and 0% (0/2) for samples obtained 12–102, 258–260, and 722–727 DPSO, respectively. Specificity for acute- and convalescent-phase samples from RT-PCR-confirmed dengue cases was 100% and 93.2%, respectively. Specificity for blood donor samples was 100%. The assay is an accurate method for Zika serological diagnosis and proved to be reliable for use during surveillance and outbreak investigations in settings where ZIKV and DENV cocirculate.

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AB0663 COMPARING METHODS FOR DETERMINING ANTIBODIES TO SARS-CoV-2 USING A RAPID TEST AND ENZYME-LINKED IMMUNOSORBENT ASSAY

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2021-eular.1311 ◽

2021 ◽

Vol 80 (Suppl 1) ◽

pp. 1363.3-1364

Author(s):

G. Gridneva ◽

E. Aronova ◽

S. Glukhova ◽

M. Cherkasova ◽

B. Belov ◽

...

Keyword(s):

Immunosorbent Assay ◽

False Negative ◽

Connective Tissue Diseases ◽

Rapid Test ◽

Test Method ◽

Enzyme Linked Immunosorbent Assay ◽

Igg Antibodies ◽

Igm Antibodies ◽

History Of

Background:An accessible and sensitive and sensitive method for determining antibodies to a new coronavirus infection is often the key to timely provision of the necessary medical care to patients with rheumatic diseasesObjectives:Compare methods for determining antibodies to SARS-CoV-2 using a rapid test and enzyme-linked immunosorbent assay (ELISA)Methods:Methods for determining antibodies to SARS-CoV-2 using an express test (Chromatographic express test SARS-CoV-2 IgG / IgM (Xiamen Biotime Biotechnology, China)) and by ELISA (Reagent kit for enzyme immunoassay of class G immunoglobulins and class M to SARS-CoV-2 (Vector-Best, Russia)) were compared.80 patients were included with a diagnosis of rheumatoid arthritis 26 (33%), psoriatic arthritis - 9 (11%), osteoarthritis - 15 (19%), rheumatic heart disease 1 (1%), SLE 2 (3%), deramtomyositis 3 (4%), systemic sclerosis 5 (6%), systemic connective tissue diseases 4 (5%), including Sjogren’s syndrome, spondyloarthritis 15 (19%).17 (21%) denied a history of COVID-19 symptoms. 63 (79%) noted any signs of COVID-19 3.095 ± 1.45 months before the test (Median 3 [2; 4] months). 63 (79%) noted any signs of COVID-19 109 ± 43 days before the test (Median 111 [78; 135] months). The ELISA method was considered the standard.Results:When comparing the results of the express test and the determination of IgG antibodies to SARS-CoV-2 in serum, the following was obtained: the sensitivity of the express test is 99%. When comparing the results of the express test and the determination of IgM antibodies to SARS-CoV-2 in serum, it was obtained: among 66 samples with a negative result by the express test method, IgM was detected in 6 cases by ELISA/ So, 7.5% of 80 samples were false negative. In 3 of 14 samples with a positive result by the express test, IgM by ELISA was not detected. So, 3.75% of 80 samples were false-positive. (Table 1). When comparing the results of the IgM express test and ELISA, the following was obtained: the sensitivity of the express test was 33%, the specificity was 85%.Table 1.Antibodies, express-testAntibodies (ELISA), absentAntibodies (ELISA), presentRowTotalsIgGabsent011Row %0.00%100.00%present37679Row %3.80%96.20%Totals37780IgМabsent60666Row %90.91%9.09%present11314Row %78.57%21.43%Totals71980Conclusion:When comparing the results of the express test and the determination of IgG antibodies to SARS-CoV-2 in serum, the sensitivity of the express test is 99%. Determination of IgM antibodies to SARS-CoV-2 using a rapid test is less reliable than determination using ELISA.Disclosure of Interests:None declared.

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SARS-CoV-2 IgG Antibodies Seroprevalence and Sera Neutralizing Activity in MEXICO: A National Cross-Sectional Study during 2020

Microorganisms ◽

10.3390/microorganisms9040850 ◽

2021 ◽

Vol 9 (4) ◽

pp. 850

Author(s):

José Esteban Muñoz-Medina ◽

Concepción Grajales-Muñiz ◽

Angel Gustavo Salas-Lais ◽

Larissa Fernandes-Matano ◽

Constantino López-Macías ◽

...

Keyword(s):

Immunosorbent Assay ◽

Neutralizing Antibodies ◽

Herd Immunity ◽

Cross Sectional Study ◽

Molecular Techniques ◽

Diagnostic Methods ◽

Molecular Diagnostic ◽

Enzyme Linked Immunosorbent Assay ◽

Cross Sectional ◽

Neutralizing Activity

Until recently, the incidence of COVID-19 was primarily estimated using molecular diagnostic methods. However, the number of cases is vastly underreported using these methods. Seroprevalence studies estimate cumulative infection incidences and allow monitoring of transmission dynamics, and the presence of neutralizing antibodies in the population. In February 2020, the Mexican Social Security Institute began conducting anonymous unrelated sampling of residual sera from specimens across the country, excluding patients with fever within the previous two weeks and/or patients with an acute respiratory infection. Sampling was carried out weekly and began 17 days before Mexico’s first officially confirmed case. The 24,273 sera obtained were analyzed by chemiluminescent-linked immunosorbent assay (CLIA) IgG S1/S2 and, later, positive cases using this technique were also analyzed to determine the rate of neutralization using the enzyme-linked immunosorbent assay (ELISA). We identified 40 CLIA IgG positive cases before the first official report of SARS-CoV-2 infection in Mexico. The national seroprevalence was 3.5% in February and 33.5% in December. Neutralizing activity among IgG positives patients during overall study period was 86.1%. The extent of the SARS-CoV-2 infection in Mexico is 21 times higher than that reported by molecular techniques. Although the general population is still far from achieving herd immunity, epidemiological indicators should be re-estimated based on serological studies of this type.

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