scholarly journals Evaluation of in-house ELISA and 11 commercial ELISA kits in the serological diagnosis of COVID-19

2021 ◽  
pp. 39-52
Author(s):  
Waldemar Rastawicki ◽  
Klaudia Płaza ◽  
Adam Pietrusiński
Keyword(s):  
Viral Infections ◽  
Clinical Symptoms ◽  
High Sensitivity ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Rt Pcr ◽  
Serum Samples ◽  
Serological Tests ◽  
Igm Antibodies ◽  
Epidemiological Surveys

Introduction: ELISA-Immunoassays can complement the molecular diagnostic methods, and can be one of the important tools of sero-surveillance and vaccine evaluation. The aim of the presented study was to develop in-house ELISA and evaluate 11 commercial ELISA tests for detection of anti-SARS-CoV-2 antibodies in serum samples collected from COVID patients. Methods: In total, 237 serum samples obtained from 165 people with COVID-19 with RT-PCR confirmed SARS-CoV-2 virus infection were used for the study. The specificity of the developed in-house ELISA kit was tested using 170 serum samples obtained from patients with various bacterial and viral infections. The study used an in-house ELISA and 11 commercial ELISA kits developed by various manufacturers. Results: The presented study showed high sensitivity (81.0%) and specificity (97.2%) of the developed in-house kit in relation to the RT-PCR method. The sensitivity of the inhouse test significantly increased (98.1%) when only convalescents - persons at least 3 weeks after COVID-19 were examined. Commercial ELISA kits most frequently detected IgG antibodies (from 44.9% to 89.4%), especially in samples obtained later in the disease, and the least frequent detection of IgM antibodies (from 4.2% to 42.4%). Conclusions: All the presented ELISA kits may be used in serodiagnosis of COVID-19 however the detection of antibodies in individual tests differed quite significantly and was dependent on the period of the disease, on the class of immunoglobulins and the type of antigen used. The sensitivity of serological tests in the IgG class is clearly higher when examining samples obtained at least 2-3 weeks from the onset of clinical symptoms. Searching for IgA antibodies may be useful mainly in the early phase of the disease while IgM antibodies does not provide significant additional information. In the case of asymptomatic or mild infection, the level of antibodies is low which may be the cause problems with the correct interpretation of epidemiological surveys

scholarly journals EVALUATION OF ELEVEN IMMUNOCHROMATOGRAPHIC ASSAYS FOR SARS-CoV-2 DETECTION: INVESTIGATING DENGUE CROSS-REACTION

2020 ◽  
Author(s):  
Beatriz Araujo Oliveira ◽  
Lea Campos de Oliveira ◽  
Franciane Mendes de Oliveira ◽  
Geovana Maria Pereira ◽  
Regina Maia de Souza ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
High Sensitivity ◽  
Diagnostic Techniques ◽  
Cross Reactivity ◽  
Cross Reaction ◽  
List Type ◽  
Rt Pcr ◽  
Serum Samples ◽  
Igm Antibodies ◽  
Good Agreement

AbstractBackgroundCOVID-19 disease (Coronavirus disease 2019) caused by SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2) is widespread worldwide, affecting more than 11 million people globally (July 6th, 2020). Diagnostic techniques have been studied in order to contain the pandemic. Immunochromatographic (IC) assays are feasible and low cost alternative for monitoring the spread of COVID-19 in the population.MethodsHere we evaluate the sensitivity and specificity of eleven different immunochromatographic tests in 98 serum samples from confirmed cases of COVID-19 through RT-PCR and 100 negative serum samples from blood donors collected in February 2019. Considering the endemic situation of Dengue in Brazil, we also evaluated the cross-reactivity with Dengue using 20 serum samples from patients with confirmed diagnosis for Dengue collected in early 2019 through four different tests.ResultsOur results demonstrated agreement between immunochromatographic assays and RT-PCR, especially after 10 days since the onset of symptoms. The evaluation of IgG and IgM antibodies combined demonstrated a strong level of agreement (0.85) of IC assays and RT-PCR. It was observed cross-reactivity between Dengue and COVID-19 using four different IC assays for COVID-19 diagnosis. The specificity of IC assays to detected COVID-19 IgM antibodies using Dengue serum samples varied from 80% to 85%; the specificity of IgG detection was 100% and total antibody was 95%.ConclusionsWe found high sensitivity, specificity and good agreement of IC assays, especially after 10 days onset of symptoms. However, we detected cross-reactivity between Dengue and COVID-19 mainly with IgM antibodies demonstrating the need for better studies about diagnostic techniques for these diseases.HighlightsImmunochromatographic assays demonstrated high sensitivity and specificity and good agreement with the gold-standard RT-PCR;Increase in sensitivity and specificity of assays using samples collected after the 10th day of symptoms;Cross-reaction with Dengue serology in evaluation of IgM.

scholarly journals Development And Performance Evaluation of A Rapid In-House ELISA for Retrospective Serosurveillance of SARS-CoV-2

2020 ◽  
Author(s):  
Bijon Kumar Sil ◽  
Mumtarin Jannat Oishee ◽  
Md. Ahsanul Haq ◽  
Nowshin Jahan ◽  
Tamanna Ali ◽  
...  
Keyword(s):  
Clinical Signs ◽  
Blocking Agent ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Rt Pcr ◽  
Serological Tests ◽  
Predictive Values ◽  
Monitoring Tools ◽  
Kappa Value ◽  
And Performance

Background In the ongoing pandemic situation of COVID-19, serological tests can complement the molecular diagnostic methods, and can be one of the important tools of sero-surveillance and vaccine evaluation. Aim To develop and evaluate a rapid SARS-CoV-2 specific IgG ELISA with increased sensitivity and specificity. Methods In order to develop the ELISA, three panels of samples (n=184) have been used: panel 1 (n=19) and panel 2 (n=60) were collected from RT-PCR positive patients within 14 and after 14 days respectively following the onset of clinical signs of disease whereas panel 3 consisted of negative samples (n=105) collected either from healthy donors during pre-pandemic era, pandemic era or from RT-PCR negative healthy individuals. As a capturing agent full-length SARS-CoV-2 specific recombinant nucleocapsid was immobilized and blocked using blocking agent. In total of 30 samples from the panels have been tested with Elecsys Anti-SARS-CoV-2 for selecting positive and negative controls, as well as comparator assay. The threshold cut-off point, inter-assay and intra assay variations were determined. Results The assay time was set at a total of 30 minutes with the sensitivity of 84% (95% confidence interval, CI, 60.4%, 96.6%) and 98% (95% CI, 91.1%, 100.0%), for panel 1 and 2 respectively, with overall 94.9% sensitivity (95% CI 87.5%, 98.6%). Moreover, the clinical specificity is 97.1% (95% CI, 91.9%, 99.4%) with no cross-reaction with dengue sample. The overall positive and negative predictive values are 96.2% (95% CI 89.2%, 99.2%) and 96.2% (95% CI, 90.6% 99.0%) respectively. In-house ELISA demonstrated 100% positive and negative percent agreement with ROCHE (Elecsys; Anti-SARS-CoV-2), with a Cohens kappa value of 1.00 (very strong agreement), while comparing 13 positive and 17 negative confirmed cases. Conclusion The assay is rapid and can be applied as one of the early and retrospective sero-monitoring tools in all over the affected areas.

scholarly journals Determination of Viral Nucleic Acid in the Human Blood

2018 ◽  
Vol 18 (4) ◽  
pp. 208-215
Author(s):  
M. A. Abdurashitov ◽  
N. A. Netesova
Keyword(s):  
Human Blood ◽  
Viral Infections ◽  
Clinical Symptoms ◽  
Viral Disease ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Reverse Transcription Reaction ◽  
Pcr Multiplex ◽  
Human Viruses

Many acute viral infections cause similar clinical symptoms, therefore, establishing the etiology of a viral disease requires the use of whole complexes of serological or PCR tests designed to detect a particular type of pathogen. Modern methods of molecular biology allow early diagnosis of viral diseases at a time when serological diagnostic methods are not yet effective. The aim of the work was to analyze molecular diagnostic methods that allow the determination of viral nucleic acids in human blood. The article presents the classification of molecular methods for the diagnosis of viral particles in clinical specimens. Methods such asin situhybridization, reverse transcription reaction (RT-PCR), nested PCR, multiplex PCR, as well as DNA microarray technology, and the method of massive parallel sequencing are considered in detail. Particular attention is paid to NGS-technologies that were used in virology almost immediately after their appearance and allowed for detection of a number of new types of human viruses (including representatives of anelloviruses, picornaviruses, polyomaviruses, etc.). The advantages and problems associated with the application of these methods in clinical practice, as well as the prospects for their improvement are discussed.

scholarly journals Development and performance evaluation of a rapid in-house ELISA for retrospective serosurveillance of SARS-CoV-2

PLoS ONE ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. e0246346
Author(s):  
Bijon Kumar Sil ◽  
Nowshin Jahan ◽  
Md. Ahsanul Haq ◽  
Mumtarin Jannat Oishee ◽  
Tamanna Ali ◽  
...  
Keyword(s):  
Clinical Symptoms ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Serological Tests ◽  
Predictive Values ◽  
Monitoring Tools ◽  
Healthy Donors ◽  
Kappa Value ◽  
And Performance ◽  
Selection Of

Background In the ongoing pandemic situation of COVID-19, serological tests can complement the molecular diagnostic methods, and can be one of the important tools of sero-surveillance and vaccine evaluation. Aim To develop and evaluate a rapid SARS-CoV-2 specific ELISA for detection of anti-SARS-CoV2 IgG from patients’ biological samples. Methods In order to develop this ELISA, three panels of samples (n = 184) have been used: panel 1 (n = 19) and panel 2 (n = 60) were collected from RT-PCR positive patients within 14 and after 14 days of onset of clinical symptoms, respectively; whereas panel 3 consisted of negative samples (n = 105) collected either from healthy donors or pre-pandemic dengue patients. As a capturing agent full-length SARS-CoV2 specific recombinant nucleocapsid was immobilized. Commercial SARS-CoV2 IgG kit based on chemiluminescent assay was used for the selection of samples and optimization of the assay. The threshold cut-off point, inter-assay and intra-assay variations were determined. Results The incubation/reaction time was set at a total of 30 minutes with the sensitivity of 84% (95% confidence interval, CI, 60.4%, 96.6%) and 98% (95% CI, 91.1%, 100.0%), for panel 1 and 2, respectively; with overall 94.9% sensitivity (95% CI 87.5%, 98.6%). Moreover, the clinical specificity was 97.1% (95% CI, 91.9%, 99.4%) with no cross reaction with dengue samples. The overall positive and negative predictive values are 96.2% (95% CI 89.2%, 99.2%) and 96.2% (95% CI, 90.6% 99.0%), respectively. In-house ELISA demonstrated 100% positive and negative percent agreement with Elecsys Anti-SARS-CoV-2, with Cohen’s kappa value of 1.00 (very strong agreement), while comparing 13 positive and 17 negative confirmed cases. Conclusion The assay is rapid and can be applied as one of the early and retrospective sero-monitoring tools in all over the affected areas.

A point-of-care diagnostic for differentiating Ebola from endemic febrile diseases

2018 ◽  
Vol 10 (471) ◽  
pp. eaat0944 ◽  
Author(s):  
David Sebba ◽  
Alexander G. Lastovich ◽  
Melody Kuroda ◽  
Eric Fallows ◽  
Joshua Johnson ◽  
...  
Keyword(s):  
Ebola Virus ◽  
Clinical Symptoms ◽  
Point Of Care ◽  
Hemorrhagic Fever ◽  
Diagnostic Methods ◽  
Outbreak Detection ◽  
Molecular Diagnostic ◽  
Clinical Samples ◽  
Live Virus ◽  
And Control

Hemorrhagic fever outbreaks such as Ebola are difficult to detect and control because of the lack of low-cost, easily deployable diagnostics and because initial clinical symptoms mimic other endemic diseases such as malaria. Current molecular diagnostic methods such as polymerase chain reaction require trained personnel and laboratory infrastructure, hindering diagnostics at the point of need. Although rapid tests such as lateral flow can be broadly deployed, they are typically not well-suited for differentiating among multiple diseases presenting with similar symptoms. Early detection and control of Ebola outbreaks require simple, easy-to-use assays that can detect and differentiate infection with Ebola virus from other more common febrile diseases. Here, we developed and tested an immunoassay technology that uses surface-enhanced Raman scattering (SERS) tags to simultaneously detect antigens from Ebola, Lassa, and malaria within a single blood sample. Results are provided in <30 min for individual or batched samples. Using 190 clinical samples collected from the 2014 West African Ebola outbreak, along with 163 malaria positives and 233 negative controls, we demonstrated Ebola detection with 90.0% sensitivity and 97.9% specificity and malaria detection with 100.0% sensitivity and 99.6% specificity. These results, along with corresponding live virus and nonhuman primate testing of an Ebola, Lassa, and malaria 3-plex assay, indicate the potential of the SERS technology as an important tool for outbreak detection and clinical triage in low-resource settings.

scholarly journals Bovine ephemeral fever epidemics in Kingdom Saudi Arabia: clinical, epidemiological and molecular investigation

2017 ◽  
Vol 11 (11) ◽  
pp. 854-860 ◽  
Author(s):  
Ahmed A Zaghawa ◽  
Fadhel Housawi ◽  
Abdulmohsen Al-Naeem ◽  
Ahmed Elsify ◽  
Yamen Mohammed Hegazy
Keyword(s):  
Saudi Arabia ◽  
Water Buffalo ◽  
Virus Isolation ◽  
Clinical Symptoms ◽  
Rt Pcr ◽  
Serum Samples ◽  
Bovine Ephemeral Fever ◽  
Geographical Regions ◽  
Epidemiological Pattern ◽  
First Time

Introduction: Bovine ephemeral fever virus (BEFV) is an arthropod borne Rhabdovirus affects cattle and water buffalo causes acute febrile disease. Methodology: The clinical picture and epidemiological pattern of BEF were described among cattle in epidemics of 2007, 2009 and 2011 in four geographical regions of Kingdom Saudi Arabia (Eastern, Jizan, Qasim, and Riyadh). Serum samples were tested using VNT. Virus isolation and molecular characterization were carried out for the first time in KSA. Results: The main clinical symptoms were fever, stiffness, lameness, salivation and subcutaneous emphysema. The prevalence and the mortality rate of BEF have decreased from 70% and 4.6% in 2007 to 30% and 0.6% in 2011, respectively in the 4 studied areas. There was no region association with higher prevalence of BEF. The intracluster correlation (ICC) was estimated for the first time in KSA as 0.0034. BEFV had been isolated from 11 out of 20 samples (55%) and isolation was confirmed by VNT. The molecular detection of BEFV by RT-PCR and real- time RT-qPCR were found more sensitive for diagnosis of the disease than virus isolation; 80% and 90% for the former tests and 55% for the latter. Three isolates were sequenced, they showed 84.7% - 100% identities in between and shared 90.4%-96.5% sequence identity with a previously published sequence from Australia (KF679404). The generated sequences belonged to 3rd cluster of BEFV glycoprotein. Conclusions: BEF occurrence has cyclic nature and the efficacy of vaccines prepared from local strains has to be evaluated and considered in diseases control.

scholarly journals Comparison of Coombs Gel Test with ELISA and Standard Tube Agglutination Tests Used in Serological Diagnosis of Brucellosis

2020 ◽  
Vol 2 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Cigdem Akalan Kuyumcu ◽  
Serpil Erol ◽  
Rıza Adaleti ◽  
Seniha Senbayrak ◽  
Secil Deniz ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Diagnostic Tests ◽  
Serological Test ◽  
High Sensitivity ◽  
Serological Diagnosis ◽  
Control Groups ◽  
Serum Samples ◽  
Serological Tests ◽  
Reliable Test ◽  
And Control

Objective: Serological tests are the most commonly used tests in the diagnosis of brucellosis; however, each serological test has some drawbacks. In this study, we aimed to determine the value of the Brucella Coombs gel test (BCGT) in the serological diagnosis of brucellosis in comparison with Standard tube agglutination (STA) and ELISA tests. Materials and Methods: The study included 42 patients who were considered to have brucellosis as a preliminary diagnosis. BCGT, Brucella-IgM/IgG ELISA, and STA tests were performed from serum samples of the patients. The correlation of the diagnostic tests was analyzed using Cohen’s Kappa Analysis.  Results: Twenty-seven (64.2%) of 42 patients were diagnosed with brucellosis according to their medical history and clinical and serological tests. The sensitivity and specificity of BCGT to diagnose brucellosis was 96.2%, and 100%, respectively. The sensitivity and specificity for the diagnosis of brucellosis 62.9% and 100% for STA, respectively; 33.3% and 66.6% for Brucella-IgM; and 66.6% and 100% for Brucella-IgG. BCGT was significantly correlated with STA (κ= 0.590) and Brucella-IgG (κ=0.539) Conclusion: BCGT can be utilized as a simple and reliable test in the diagnosis of brucellosis with high sensitivity and specificity. Nevertheless, the sensitivity and specificity of BCGT should be demonstrated by comprehensive studies, including culture-confirmed cases and control groups.

scholarly journals Leishmania (infantum) chagasi in canine urinary sediment

2015 ◽  
Vol 24 (1) ◽  
pp. 92-94 ◽  
Author(s):  
Ivete Lopes de Mendonça ◽  
Joilson Ferreira Batista ◽  
Leucio Camara Alves
Keyword(s):  
Bone Marrow ◽  
Clinical Symptoms ◽  
High Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Canine Visceral Leishmaniasis ◽  
Serological Tests ◽  
Urine Sediment ◽  
Urinary Sediment ◽  
Control Center ◽  
Veterinary Hospital

Canine visceral leishmaniasis (CVL) is difficult to diagnosis, mainly due to the presence of asymptomatic animals, the diversity of clinical symptoms and the difficulty in obtaining diagnostic evidence of high sensitivity and specificity. The purpose of this study was to diagnose CVL in urinary sediment of 70 dogs of different breeds, sexes and ages from the veterinary hospital of the Federal University of Piauí and Zoonosis Control Center of Teresina, Brazil. The serological tests were TR DPP® for CVL and enzyme-linked immunosorbent assay (ELISA) for CVL, parasitological exams of bone marrow and lymph nodes and urine sediment cultures. Leishmania was detected in the bone marrow and/or lymph node of 61.0% of the animals (43/70), and urine sediment culture was positive in 9.30% (4/43) of these animals. In the serological exams, 70.0% (49/70) were reactive using the DPP and 78.2% (55/70) were reactive using ELISA. The goal of this study was to diagnose the presence of L. (infantum) chagasi in a culture of urinary sediment.

scholarly journals Point-of-Care Diagnostic Tools for Surveillance of SARS-CoV-2 Infections

2021 ◽  
Vol 9 ◽  
Author(s):  
Dhanasekaran Sakthivel ◽  
David Delgado-Diaz ◽  
Laura McArthur ◽  
William Hopper ◽  
Jack S. Richards ◽  
...  
Keyword(s):  
Large Scale ◽  
Clinical Symptoms ◽  
Point Of Care ◽  
Cost Effective ◽  
Nucleic Acid Amplification ◽  
Diagnostic Tools ◽  
Rt Pcr ◽  
Serological Tests ◽  
Polymerase Chain ◽  
Effective Public Health

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a recently emerged and highly contagious virus that causes coronavirus disease 2019 (COVID-19). As of August 24, 2021, there were more than 212 million confirmed COVID-19 cases and nearly 4.4 million deaths reported globally. Early diagnosis and isolation of infected individuals remains one of the most effective public health interventions to control SARS-CoV-2 spread and for effective clinical management of COVID-19 cases. Currently, SARS-CoV-2 infection is diagnosed presumptively based on clinical symptoms and confirmed by detecting the viral RNA in respiratory samples using reverse transcription polymerase chain reaction (RT-PCR). Standard RT-PCR protocols are time consuming, expensive, and technically demanding, which makes them a poor choice for large scale and point-of-care screening in resource-poor settings. Recently developed isothermal nucleic acid amplification tests (iNAAT), antigen and/or serological tests are cost-effective to scale COVID-19 testing at the point-of-care (PoC) and for surveillance activities. This review discusses the development of rapid PoC molecular tools for the detection and surveillance of SARS-CoV-2 infections.

scholarly journals Evaluation of nine serological rapid tests for the detection of SARS-CoV-2

2020 ◽  
Vol 44 ◽  
pp. 1
Author(s):  
Marcela Mercado ◽  
Jeadran Malagón-Rojas ◽  
Gabriela Delgado ◽  
Vivian Vanesa Rubio ◽  
Lida Muñoz Galindo ◽  
...  
Keyword(s):  
Study Groups ◽  
Cross Sectional Study ◽  
Igg Antibodies ◽  
Sectional Study ◽  
Rt Pcr ◽  
Serum Samples ◽  
Serological Tests ◽  
Cross Sectional ◽  
Diagnostic Aid ◽  
Rapid Tests

Objective. To evaluate the operative capacity of nine serological rapid tests to detect the IgM/IgG antibodies response in serum from patients with SARS-CoV-2 in different clinical stages. Methods. A cross-sectional study of serological rapid tests was designed to compare the performance of the evaluated immunochromatographic tests for the diagnosis of SARS-CoV-2. A total of 293 samples was used, including negatives, asymptomatic, and symptomatic serum samples. Results. The sensitivity of the evaluated tests was low and moderate in the groups of asymptomatic serum samples and the group of serums coming from patients with less than 11 days since the onset of the symptoms. The specificity for the anti-SARS-CoV-2 antibodies tests ranged between 86.5%-99% for IgM and 86.5%-99.5% for IgG. The sensitivity and the likelihood ratio were different according to the study groups. The usefulness of these tests is restricted to symptomatic patients and their sensitivity is greater than 85% after 11 days from the appearance of symptoms. Conclusions. Serological tests are not an adequate strategy for the identification of asymptomatic and pre-symptomatic patients. Serological rapid tests for the detection of specific anti-SARS-CoV-2 antibodies can be used as a diagnostic aid, but diagnosis must be confirmed by RT-PCR. Rapid tests should be reserved for patients with symptoms lasting more than 11 days.

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