scholarly journals Comparison of Two Rapid Diagnostic Assays for Detection of Immunoglobulin M Antibodies to Dengue Virus

2000 ◽  
Vol 7 (1) ◽  
pp. 106-110 ◽  
Author(s):  
Shuenn-Jue L. Wu ◽  
Helene Paxton ◽  
Barbara Hanson ◽  
Cheryl G. Kung ◽  
Timothy B. Chen ◽  
...  
Keyword(s):  
Dengue Virus ◽  
False Positive ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Detection ◽  
Diagnostic Assays ◽  
Igm Antibody ◽  
Elisa Format ◽  
Assay Time ◽  
Antibody Capture

ABSTRACT Two easy-to-use commercial diagnostic assays, a dipstick enzyme-linked immunosorbent assay (ELISA) (Integrated Diagnostics, Baltimore, Md.) and an immunochromatographic card assay (PanBio, Brisbane, Australia) were evaluated for detection of immunoglobulin M (IgM) antibody to dengue virus with an in-house IgM antibody capture microplate ELISA as a reference assay. The dipstick ELISA was based on the indirect-ELISA format using dengue 2 virus as the only antigen and enzyme-labeled goat anti-human IgM antibody as the detector. The total assay time was 75 min. The immunochromatographic card assay was based on the antibody capture format and separately measured both anti-dengue virus IgM and IgG in the same test. Colloidal-gold-labeled anti-dengue virus monoclonal antibody bound with dengue virus 1 to 4 antigen cocktail was the detector, and anti-human IgM and IgG were the capture antibodies. The total assay time was <10 min. Sera from 164 individuals classified as either anti-dengue virus IgM positive (94) or anti-dengue virus IgM negative (70) in the reference microplate ELISA with a dengue virus 1 to 4 antigen cocktail were tested in the two commercial assays. The dipstick ELISA missed 7 of 94 positive samples, for a sensitivity of 92.6%, while the immunochromatographic card assay missed two positive samples, for a sensitivity of 97.9%. Of the 70 negative samples, four were false positive by the dipstick ELISA and two were false positive in the immunochromatographic card assay, resulting in specificities of 94.3 and 97.1%, respectively. Both commercial assays provide sensitive and specific detection of anti-dengue virus IgM antibody and could prove useful in settings where the microplate ELISA is impractical.

scholarly journals Comparison of Seven Commercial Antigen and Antibody Enzyme-Linked Immunosorbent Assays for Detection of Acute Dengue Infection

2012 ◽  
Vol 19 (5) ◽  
pp. 804-810 ◽  
Author(s):  
Stuart D. Blacksell ◽  
Richard G. Jarman ◽  
Robert V. Gibbons ◽  
Ampai Tanganuchitcharnchai ◽  
Mammen P. Mammen ◽  
...  
Keyword(s):  
South Korea ◽  
Dengue Virus ◽  
Dengue Infection ◽  
Virus Antigen ◽  
Enzyme Linked Immunosorbent Assay ◽  
Capture Elisa ◽  
Igm Antibody ◽  
Antigen Elisa ◽  
Ns1 Antigen ◽  
Elisa Format

ABSTRACTSeven commercial assays were evaluated to determine their suitability for the diagnosis of acute dengue infection: (i) the Panbio dengue virus Pan-E NS1 early enzyme-linked immunosorbent assay (ELISA), second generation (Alere, Australia); (ii) the Panbio dengue virus IgM capture ELISA (Alere, Australia); (iii) the Panbio dengue virus IgG capture ELISA (Alere, Australia); (iv) the Standard Diagnostics dengue virus NS1 antigen ELISA (Standard Diagnostics, South Korea); (v) the Standard Diagnostics dengue virus IgM ELISA (Standard Diagnostics, South Korea); (vi) the Standard Diagnostics dengue virus IgG ELISA (Standard Diagnostics, South Korea); and (vii) the Platelia NS1 antigen ELISA (Bio-Rad, France). Samples from 239 Thai patients confirmed to be dengue virus positive and 98 Sri Lankan patients negative for dengue virus infection were tested. The sensitivities and specificities of the NS1 antigen ELISAs ranged from 45 to 57% and 93 to 100% and those of the IgM antibody ELISAs ranged from 85 to 89% and 88 to 100%, respectively. Combining the NS1 antigen and IgM antibody results from the Standard Diagnostics ELISAs gave the best compromise between sensitivity and specificity (87 and 96%, respectively), as well as providing the best sensitivity for patients presenting at different times after fever onset. The Panbio IgG capture ELISA correctly classified 67% of secondary dengue infection cases. This study provides strong evidence of the value of combining dengue virus antigen- and antibody-based test results in the ELISA format for the diagnosis of acute dengue infection.

scholarly journals Development of a Human-Murine Chimeric Immunoglobulin M Antibody for Use in the Serological Detection of Human Flavivirus Antibodies

2009 ◽  
Vol 16 (5) ◽  
pp. 679-685 ◽  
Author(s):  
Brett A. Thibodeaux ◽  
John T. Roehrig
Keyword(s):  
Yellow Fever Virus ◽  
Positive Control ◽  
Encephalitis Virus ◽  
Immunoglobulin M ◽  
Chimeric Antibody ◽  
Enzyme Linked Immunosorbent Assay ◽  
Variable Regions ◽  
Igm Antibody ◽  
Antibody Capture ◽  
Serological Activity

ABSTRACT Current diagnosis of human flaviviral infections relies heavily on serological techniques such as the immunoglobulin M (IgM) antibody capture enzyme-linked immunosorbent assay (MAC-ELISA). Broad application of this assay is hindered by a lack of standardized human positive-control sera that react with the wide variety of flaviviruses that can cause human disease, e.g., dengue virus (DENV), West Nile virus (WNV), yellow fever virus (YFV), Japanese encephalitis virus (JEV), and St. Louis encephalitis virus (SLEV). We have created a human-murine chimeric antibody combining the variable regions of the broadly flavivirus cross-reactive murine monoclonal antibody (MAb) 6B6C-1 and the constant region of human IgM to produce a standardized reagent capable of replacing human positive-control sera in a MAC-ELISA for the diagnosis of all human flaviviral infections. The human-murine chimeric IgM antibody secreted from plasmid-transformed Sp2/0-Ag14 cells had a level of serological activity identical to that of 6B6C-1 as measured by ELISA, immunoblotting, and MAC-ELISA for multiple members of the flavivirus genus, including WNV, SLEV, YFV, DENV, and JEV.

scholarly journals Serotype-Cross-Reactive Immunoglobulin M Responses in Dengue Virus Infections Determined by Enzyme-Linked Immunosorbent Assay

2000 ◽  
Vol 7 (5) ◽  
pp. 774-777 ◽  
Author(s):  
Masaru Nawa ◽  
Ken-Ichiro Yamada ◽  
Tomohiko Takasaki ◽  
Toshitaka Akatsuka ◽  
Ichiro Kurane
Keyword(s):  
Dengue Virus ◽  
Japanese Patients ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Infections ◽  
Serum Samples ◽  
Reverse Transcription Pcr ◽  
Virus Serotype ◽  
Dengue Virus Serotypes ◽  
Antibody Capture

ABSTRACT We developed immunoglobulin M (IgM) antibody capture enzyme-linked immunosorbent assays (ELISAs) with four monovalent dengue virus antigens. We attempted to determine whether IgM responses in dengue virus infections are serotype specific or serotype cross-reactive. Serum samples from 14 confirmed dengue cases were examined. In these 14 cases, which consisted of 12 Japanese and 2 non-Japanese patients, infecting dengue virus serotypes were defined by reverse transcription-PCR. Thirteen of the 14 cases were IgM positive in ELISA. IgM responses were serotype cross-reactive in these 13 cases but were highest against infecting dengue virus serotype in 9 of the 13 cases. These results indicate that IgM responses are generally dengue serotype cross-reactive but that IgM levels are highest against the infecting serotype in most dengue cases.

scholarly journals Development of a More Efficient Algorithm for Identifying False-Positive Reactivity Results in a Dengue Virus Immunoglobulin M Screening Assay

2008 ◽  
Vol 15 (8) ◽  
pp. 1304-1306 ◽  
Author(s):  
Harry E. Prince ◽  
Cindy Yeh ◽  
Mary Lapé-Nixon
Keyword(s):  
Dengue Virus ◽  
Background Subtraction ◽  
False Positive ◽  
Efficient Algorithm ◽  
False Positives ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Screening Assay ◽  
Positive Reactivity ◽  
Positive Index

ABSTRACT Of 2,692 sera screened for dengue virus immunoglobulin M by using a μ-capture enzyme-linked immunosorbent assay (ELISA), 954 had equivocal (index from 0.90 to 1.10) or positive (index of >1.10) results and were retested using a background subtraction (BS) ELISA that identifies screen false positives. No false positives were found among 427 sera with screen ELISA indices of >6.00; thus, retesting this specimen subset by BS ELISA is unnecessary.

scholarly journals Diagnosis of Dengue Virus Infection by Detection of Specific Immunoglobulin M (IgM) and IgA Antibodies in Serum and Saliva

2003 ◽  
Vol 10 (2) ◽  
pp. 317-322 ◽  
Author(s):  
Angel Balmaseda ◽  
María G. Guzmán ◽  
Samantha Hammond ◽  
Guillermo Robleto ◽  
Carolina Flores ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Immunoglobulin M ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Serum Samples ◽  
Diagnostic Modality ◽  
Iga Antibodies ◽  
Specific Immunoglobulin ◽  
Feasible Alternative

ABSTRACT To evaluate alternative approaches to the serological diagnosis of dengue virus (DEN) infection, the detection of DEN-specific immunoglobulin M (IgM) and IgA antibodies in serum and saliva specimens was assessed in 147 patients with symptoms of DEN infection seen at the Ministry of Health in Nicaragua. Seventy-two serum samples were determined to be positive for anti-DEN antibodies by IgM capture enzyme-linked immunosorbent assay, the routine diagnostic procedure. Serum and saliva specimens were obtained from 50 healthy adults as additional controls. IgM was detected in the saliva of 65 of the 72 serum IgM-positive cases, 6 of the 75 serum IgM-negative cases, and none of the control group, resulting in a sensitivity of 90.3% and a specificity of 92.0% and demonstrating that salivary IgM is a useful diagnostic marker for DEN infection. Detection of IgA in serum may be another feasible alternative for the diagnosis of DEN infection, with serum IgA found in 68 (94.4%) of the IgM-positive cases. In contrast, detection of IgA in saliva was not found to be a useful tool for DEN diagnosis in the present study. Further studies of the kinetics of antibody detection in another set of 151 paired acute- and convalescent-phase serum samples showed that DEN-specific IgA antibodies were detected in more acute-phase samples than were IgM antibodies. Thus, we conclude that DEN-specific IgA in serum is a potential diagnostic target. Furthermore, given that saliva is a readily obtainable, noninvasive specimen, detection of DEN-specific salivary IgM should be considered a useful, cheaper diagnostic modality with similar sensitivity and specificity to IgM detection in serum.

The Development and Interlaboratory Validation of a Quantitative Antibody Capture ELISA for the Measurement of Specific Guinea-pig IgG1: An Alternative to the Passive Cutaneous Anaphylaxis Assay

1996 ◽  
Vol 24 (1) ◽  
pp. 73-79
Author(s):  
Laura S. Babcock ◽  
Thomas T. Kawabata ◽  
Victor S. Moore ◽  
Catherine M. Condo ◽  
Annette Prentø
Keyword(s):  
Guinea Pig ◽  
Passive Cutaneous Anaphylaxis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Antigens ◽  
Capture Elisa ◽  
Elisa Format ◽  
Interlaboratory Validation ◽  
Antibody Capture ◽  
Development And Validation

Specific guinea-pig IgG1 has traditionally been measured by using an in vivo guinea-pig passive cutaneous anaphylaxis (PCA) assay. This paper describes the development and validation of a quantitative enzyme-linked immunosorbent assay (ELISA) for specific IgG1 against two protein antigens (Alcalase and Enzyme B) as an alternative to the PCA assay. The ELISA format involved a rabbit antibody bound to microtitre plates to capture the antigen. The test sera is added to this, followed sequentially by goat anti-guinea-pig IgG1 and rabbit anti-goat-IgG-alkaline phosphatase conjugate. Aliquots of each serum sample from immunised guinea-pigs were analysed with the ELISA for specific IgG1 titres in three laboratories and were compared to titres determined by using the PCA assay. The findings demonstrate that there is a good correlation between the ELISA and the PCA assay and that the ELISA shows good interlaboratory reproducibility. Thus, the antibody capture ELISA described in this report is a valid and robust replacement for the guinea-pig PCA assay.

scholarly journals Immunoglobulin A Antibody Responses in Dengue Patients: a Useful Marker for Serodiagnosis of Dengue Virus Infection

2005 ◽  
Vol 12 (10) ◽  
pp. 1235-1237 ◽  
Author(s):  
M. Nawa ◽  
T. Takasaki ◽  
M. Ito ◽  
S. Inoue ◽  
K. Morita ◽  
...  
Keyword(s):  
Virus Infection ◽  
Dengue Virus ◽  
Immunoglobulin A ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Infections ◽  
Serum Samples ◽  
Dengue Virus Infection ◽  
Iga Antibody ◽  
Antibody Capture

ABSTRACT We determined the usefulness of an immunoglobulin A (IgA) antibody-capture enzyme-linked immunosorbent assay for serodiagnosis of dengue virus infections. The results indicate that the presence of IgA and IgM in serum samples assures recent primary dengue virus infection even with a single serum sample.

scholarly journals Technical viability of the YF MAC-HD ELISA kit for use in yellow fever-endemic regions

2021 ◽  
Vol 15 (6) ◽  
pp. e0009417
Author(s):  
Christin H. Goodman ◽  
Maurice Demanou ◽  
Mick Mulders ◽  
Jairo Mendez-Rico ◽  
Alison Jane Basile
Keyword(s):  
South America ◽  
Yellow Fever ◽  
Accurate Diagnosis ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Kit ◽  
Wash Buffer ◽  
Antibody Capture ◽  
Vaccination Campaigns ◽  
Arboviral Disease

Yellow fever (YF), an arboviral disease, affects an estimated 200,000 people and causes 30,000 deaths per year and recently has caused major epidemics in Africa and South America. Timely and accurate diagnosis of YF is critical for managing outbreaks and implementing vaccination campaigns. A YF immunoglobulin M (IgM) antibody-capture (MAC) enzyme-linked immunosorbent assay (ELISA) kit, the YF MAC-HD, was successfully introduced starting in 2018 to laboratories in Africa and South America. The YF MAC-HD kit can be performed in 3.5 hours, test up to 24 samples, and includes all reagents necessary to perform the test, except for water used to dilute wash buffer. In 2018 and 2019, a total of 56 laboratory personnel from 39 countries in Africa and South America were trained to use the kit during workshops, followed by take-home YF IgM proficiency testing (PT) exercises. Participants received either a 10- or 20-sample YF PT panel and performed testing using the YF MAC-HD kit. All countries obtained 90% or higher correct results. These results verified the technical viability and transferability of YF MAC-HD kit use for laboratories in YF-endemic countries.

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.

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