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 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 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.

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.

scholarly journals Evaluation of a Novel Reporter Virus Neutralization Test for Serological Diagnosis of Zika and Dengue Virus Infection

2017 ◽  
Vol 55 (10) ◽  
pp. 3028-3036 ◽  
Author(s):  
Chao Shan ◽  
Daniel A. Ortiz ◽  
Yujiao Yang ◽  
Susan J. Wong ◽  
Laura D. Kramer ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Neutralizing Antibodies ◽  
Neutralization Test ◽  
Reference Method ◽  
Laboratory Diagnosis ◽  
Virus Neutralization Test ◽  
Virus Neutralization ◽  
Enzyme Linked Immunosorbent Assay ◽  
Reporter Virus ◽  
Screening Assay

ABSTRACT Currently, the laboratory diagnosis of Zika virus (ZIKV) infection is primarily through the detection of ZIKV RNA or antibodies against ZIKV proteins. The detection of viral RNA is highly sensitive and specific, but periods of viremia and viruria are brief, limiting the utility of ZIKV RNA assays. Instead, most ZIKV infections are diagnosed serologically, using an IgM antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) for screening, followed by a confirmatory plaque reduction neutralization test (PRNT). Typical turnaround times vary, due to assay incubation periods and a lack of clinical laboratories performing these tests. Recently, a novel luciferase-ZIKV- and -dengue virus (DENV)-based serological assay, which considerably improves the turnaround times and throughput for ZIKV diagnosis, was described. Using the traditional PRNT as a reference method, we evaluated the performance characteristics of the reporter virus neutralization test (RVNT) with 258 clinical serum specimens. The ZIKV RVNT produced primary ZIKV screening and secondary confirmation results in 4 days, with 100% reproducibility. As a screening assay, the ZIKV RVNT displayed excellent diagnostic accuracy, sensitivity, and specificity of 98.2%, 100%, and 98.1%, respectively. As a confirmatory assay, the ZIKV RVNT titers displayed 93.1% agreement with the traditional ZIKV PRNT titers. Overall, the RVNT accurately and reliably detects neutralizing antibodies in patient serum specimens, with improved turnaround times, and can be used for the serological detection of ZIKV infections. Due to the homogeneous 96-well format, the RVNT has also significantly improved the assay throughput to allow testing of a large number of specimens in a single run.

scholarly journals A Method To Prevent SARS-CoV-2 IgM False Positives in Gold Immunochromatography and Enzyme-Linked Immunosorbent Assays

2020 ◽  
Vol 58 (6) ◽  
Author(s):  
Qiang Wang ◽  
Qin Du ◽  
Bin Guo ◽  
Daiyong Mu ◽  
Xiaolan Lu ◽  
...  
Keyword(s):  
Legionella Pneumophila ◽  
False Positive ◽  
Influenza A ◽  
Influenza B Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Influenza B ◽  
Content Type ◽  
Positive Reactivity ◽  
Positive Serum ◽  
High Level

ABSTRACT We set out to investigate the interference factors that led to false-positive novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) IgM detection results using gold immunochromatography assay (GICA) and enzyme-linked immunosorbent assay (ELISA) and the corresponding solutions. GICA and ELISA were used to detect SARS-CoV-2 IgM in 86 serum samples, including 5 influenza A virus (Flu A) IgM-positive sera, 5 influenza B virus (Flu B) IgM-positive sera, 5 Mycoplasma pneumoniae IgM-positive sera, 5 Legionella pneumophila IgM-positive sera, 6 sera of HIV infection patients, 36 rheumatoid factor IgM (RF-IgM)-positive sera, 5 sera from hypertensive patients, 5 sera from diabetes mellitus patients, and 14 sera from novel coronavirus infection disease 19 (COVID-19) patients. The interference factors causing false-positive reactivity with the two methods were analyzed, and the urea dissociation test was employed to dissociate the SARS-CoV-2 IgM-positive serum using the best dissociation concentration. The two methods detected positive SARS-CoV-2 IgM in 22 mid-to-high-level-RF-IgM-positive sera and 14 sera from COVID-19 patients; the other 50 sera were negative. At a urea dissociation concentration of 6 mol/liter, SARS-CoV-2 IgM results were positive in 1 mid-to-high-level-RF-IgM-positive serum and in 14 COVID-19 patient sera detected using GICA. At a urea dissociation concentration of 4 mol/liter and with affinity index (AI) levels lower than 0.371 set to negative, SARS-CoV-2 IgM results were positive in 3 mid-to-high-level-RF-IgM-positive sera and in 14 COVID-19 patient sera detected using ELISA. The presence of RF-IgM at mid-to-high levels could lead to false-positive reactivity of SARS-CoV-2 IgM detected using GICA and ELISA, and urea dissociation tests would be helpful in reducing SARS-CoV-2 IgM false-positive results.

scholarly journals KrakenHLL: Confident and fast metagenomics classification using unique k-mer counts

2018 ◽  
Author(s):  
FP Breitwieser ◽  
SL Salzberg
Keyword(s):  
Infectious Disease ◽  
False Positive ◽  
Efficient Algorithm ◽  
False Positives ◽  
Significant Problem ◽  
Additional Memory

AbstractFalse positive identifications are a significant problem in metagenomic classification. We present KrakenHLL, a novel metagenomic classifier that combines the fast k-mer based classification of Kraken with an efficient algorithm for assessing the coverage of unique k-mers found in each species in a dataset. On various test datasets, KrakenHLL gives better recall and F1-scores than other methods, and effectively classifies and distinguishes pathogens with low abundance from false positives in infectious disease samples. By using the probabilistic cardinality estimator HyperLogLog (HLL), KrakenHLL is as fast as Kraken and requires little additional memory.

scholarly journals Co-circulation of all the Four Serotype of Dengue Virus in Endemic Region of Saurashtra, Gujarat during 2019-2020 Season

2021 ◽  
Author(s):  
Binita Joseph Aring ◽  
Dipali Magan Bhai Gavali ◽  
Pushpa Ramjibhai Kateshiya ◽  
Hiral Modbhai Gadhvi ◽  
Summaiya Mullan ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Nonstructural Protein ◽  
Geographical Area ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Endemic Region ◽  
Male Population ◽  
Monsoon Period ◽  
Cross Sectional ◽  
Nonstructural Protein 1

Introduction: Dengue has rapidly emerged as a vector - borne viral disease in recent years and also endemic in all continents. The agent of dengue, i.e., dengue viruses, are categorised under the genus Flavivirus, with the four dengue virus serotypes: designated as DENV-1, DENV-2, DENV-3 and DENV-4. These all four serotypes are in circulation either singly, or more than one at the same time. Aim: To study the epidemiological update of dengue with circulating serotype and co-infection in Saurashtra region, Gujarat, India. Materials and Methods: A cross-sectional study was conducted during January 2019 to December 2020 and total samples received were 12,563 which were clinically suspected dengue samples case. After receiving blood samples, serum was separated and proceeded for Dengue NS1Ag (nonstructural protein 1 antigen), and Dengue IgM Ab (Immunoglobulin M antibody). After serological confirmation, 151 samples from different geographical area were selected for Dengue specific Reverse Transcriptase- Polymerase Chain Reaction (RT-PCR) for serotyping. results: Total 4069 (32%) had confirmed dengue positive by Enzyme Linked Immunosorbent Assay (ELISA). The ratio of male cases was higher than female, and in age group 21-35 year (47%). Seasonal trend showed a gradual increase in positivity from June with high peak in October. Circulation of all the four serotypes in area, higher monotypic infection by DENV-1 serotype (41%), followed by DENV-4, DENV-2 and DENV-3. Co-infection of different serotypes were also found. conclusion: The present study concluded that all four serotypes circulate with predominant being DENV-1 type and co-infection of different serotypes in the saurashtra region. Dengue mainly affected adult male population, and seasonal peak during monsoon and post-monsoon period.

Immunoglobulin G- and M-specific enzyme-linked immunosorbent assay for detection of dengue antibodies

1979 ◽  
Vol 9 (4) ◽  
pp. 498-502
Author(s):  
D Dittmar ◽  
T J Cleary ◽  
A Castro
Keyword(s):  
Dengue Virus ◽  
Immunosorbent Assay ◽  
Immunoglobulin G ◽  
Hemagglutination Inhibition ◽  
Dengue Infection ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Enzyme ◽  
Enzyme Conjugate ◽  
Inhibition Technique

An enzyme-linked immunosorbent assay for the detection of antibodies to dengue virus is described. This method correlates well with a hemagglutination inhibition technique. The enzyme-linked immunosorbent assay can also be specific for human immunoglobulin M antibodies when a mu-chain-specific antiglobulin-enzyme conjugate and fractionated serum are employed. By using this technique, dengue immunoglobulin M antibodies were demonstrated in an infant suspected of having a recent dengue infection.

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 Prospective Study To Determine Accuracy of Rapid Serological Assays for Diagnosis of Acute Dengue Virus Infection in Laos

2007 ◽  
Vol 14 (11) ◽  
pp. 1458-1464 ◽  
Author(s):  
Stuart D. Blacksell ◽  
David Bell ◽  
James Kelley ◽  
Mammen P. Mammen ◽  
Robert V. Gibbons ◽  
...  
Keyword(s):  
Virus Infection ◽  
Dengue Virus ◽  
Patient Management ◽  
Limited Resource ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Infections ◽  
Serum Samples ◽  
Dengue Virus Infection ◽  
Resource Setting

ABSTRACT There is an urgent need for accurate and simple dengue virus infection diagnostic assays in limited-resource settings of dengue endemicity, to assist patient management. Using a panel of reference samples (S. D. Blacksell, P. N. Newton, D. Bell, J. Kelley, M. P. Mammen, D. W. Vaughn, V. Wuthiekanun, A. Sungkakum, A. Nisalak, and N. P. Day, Clin. Infect. Dis. 42:1127-1134, 2006), we recently evaluated eihgt commercially available immunochromatographic rapid diagnostic tests (RDTs) designed to detect dengue virus-specific immunoglobulin M (IgM) and/or IgG. We found that 6/8 RDTs had sensitivities of less than 50% (range, 6 to 65%), but specificities were generally high. Here, in conjuction with dengue virus serotyping by reverse transcriptase PCR and in the limited-resource setting of Laos, where dengue virus is endemic, we evaluated the same eight RDTs against a previously validated dengue IgM/IgG enzyme-linked immunosorbent assay for diagnosis of acute dengue virus infection. Paired serum samples were collected from 87 patients, of whom 38 had confirmed dengue virus infections (4 had primary infections, 33 had secondary infections, and 1 had an infection of indeterminate status). RDT sensitivity was low, with 7/8 RDTs having admission sample sensitivities of less than 20% (range, 4 to 26%). The majority (6/8) of the RDTs, demonstrated high specificity (>95%). Kappa statistic values ranged from 6 to 54% for the RDTs, demonstrating poor to moderate variation between three operators. No RDT adequately differentiated between primary and secondary dengue virus infections. The findings of this study suggest that currently available RDTs based on the detection of IgM antibodies for the diagnosis of acute dengue virus infections are unlikely to be useful for patient management.

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