Dataset Information

Development and validation of different indirect ELISAs for MERS-CoV serological testing.

ABSTRACT: Since 2012, MERS-CoV has caused up to 2220 cases and 790 deaths in 27 countries with Saudi Arabia being the most affected country with ~83.1% of the cases and ~38.8% local death rate. Current serological assays such as microneutralization (MN), plaque reduction neutralization, immunofluorescence, protein microarray or pseudoparticle neutralization assays rely on handling of live MERS-CoV in high containment laboratories or need for expensive and special equipment and reagents and highly trained personnel which represent a technical hurdle for most laboratories in resource-limited MERS-CoV endemic countries. Here, we developed, compared and evaluated three different indirect ELISAs based on MERS-CoV nucleocapsid protein (N), spike (S) ectodomain (amino acids 1-1297) and S1 subunit (amino acids 1-725) and compared them with MN assay. The developed ELISAs were evaluated using large number of confirmed seropositive (79 samples) and seronegative (274 samples) MERS-CoV human serum samples. Both rS1- and rS-ELISAs maintained high sensitivity and specificity (?90%) across a wider range of OD values compared to rN-ELISA. Moreover, rS1- and rS-based ELISAs showed better agreement and correlation with MN assay in contrast to rN-ELISA. Collectively, our data demonstrate that rS1-ELISA and rS-ELISA are more reliable than rN-ELISA and represent a suitable choice for seroepidemiological testing and surveillance in MERS-CoV endemic regions.

SUBMITTER: Hashem AM

PROVIDER: S-EPMC7094657 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Development and validation of different indirect ELISAs for MERS-CoV serological testing.

Hashem Anwar M AM Al-Amri Sawsan S SS Al-Subhi Tagreed L TL Siddiq Loai A LA Hassan Ahmed M AM Alawi Maha M MM Alhabbab Rowa Y RY Hindawi Salwa I SI Mohammed Osama B OB Amor Nabil S NS Alagaili Abdulaziz N AN Mirza Ahmed A AA Azhar Esam I EI

Journal of immunological methods 20190116

Since 2012, MERS-CoV has caused up to 2220 cases and 790 deaths in 27 countries with Saudi Arabia being the most affected country with ~83.1% of the cases and ~38.8% local death rate. Current serological assays such as microneutralization (MN), plaque reduction neutralization, immunofluorescence, protein microarray or pseudoparticle neutralization assays rely on handling of live MERS-CoV in high containment laboratories or need for expensive and special equipment and reagents and highly trained ...[more]

PMID: 30659836

Similar Datasets

Project description:COVID-19 is the name of the acute respiratory disease caused by the new coronavirus SARS-CoV-2, a close relative of those that caused the severe outbreaks of SARS and MERS several years ago. Since first appearance on December of 2019, the COVID-19 pandemic has cause extremely high levels of mortality, morbidity, global economic breakdown, and the consequent human suffering. The main diagnostic test for the confirmation of symptomatic individuals is the detection of viral RNA by reverse transcriptase-quantitative real time PCR (RT-PCR). Additionally, serology techniques, such as ELISA are useful to measure the antibodies produced in humans after contact with the virus, as well as the direct presence of viral antigens. In this study we aim to assemble and evaluate four ELISA assays to measure the presence of IgG or IgM specific for the viral Spike protein in COVID-19 patients, using either the full recombinant SARS-CoV-2 Spike protein or the fragment corresponding to the receptor binding domain. As a control, we analyzed a group of pre-pandemic serum samples obtained before 2017. Strong reactivity was observed against both antigens. A few pre-pandemic samples displayed high OD values, suggesting the possibility of some cross reactivity. All four assays show very good repeatability, both intra- and inter-assay. Receiver operating characteristic analysis allowed the definition of cutoffs and evaluation of performance for each ELISA by estimation of the area under the curve. This performance parameter was high for all tests (AUC range: 0.98-0.99). Multiple comparisons between tests revealed no significant difference between each other (P values: 0.24-0.95). Our results show that both antigens are effective to detect both specific IgG and IgM antibodies, with high sensitivity (range 0.92-0.99), specificity (range 0.93-0.97) and congruence with the RT-PCR test (Cohen´s Kappa range 0.87-0.93). These assays will allow health authorities to have a new tool to estimate seroprevalence, in order to manage and improve the severe sanitary situation caused by this virus.

Project description:Middle East Respiratory Syndrome Coronavirus (MERS-CoV) is an emerging viral disease and dromedary camels are known to be the source of human spill over events. A cross-sectional epidemiological surveillance study was carried out in Kenya in 2017 to, 1) estimate MERS-CoV antibody seropositivity in the camel-dense counties of Turkana, Marsabit, Isiolo, Laikipia and Nakuru to identify, and 2) determine the risk factors associated with seropositivity in camels. Blood samples were collected from a total of 1421 camels selected using a multi-stage sampling method. Data were also collected from camel owners or herders using a pre-tested structured questionnaire. The sera from camel samples were tested for the presence of circulating antibodies to MERS-CoV using the anti-MERS-CoV IgG ELISA test. Univariate and multivariable statistical analysis were used to investigate factors potentially associated with MERS-CoV seropositivity in camels. The overall seropositivity in camel sera was 62.9 %, with the highest seropositivity recorded in Isiolo County (77.7 %), and the lowest seropositivity recorded in Nakuru County (14.0 %). When risk factors for seropositivity were assessed, the "Type of camel production system" {(aOR = 5.40(95 %CI: 1.67-17.49)}, "Age between 1-2 years, 2-3 years and above 3 years" {(aOR = 1.64 (95 %CI: 1.04-2.59}", {(aOR = 3.27 (95 %CI: 3.66-5.61)}" and {(aOR = 6.12 (95 %CI: 4.04-9.30)} respectively and "Sex of camels" {(aOR = 1.75 (95 %CI: 1.27-2.41)} were identified as significant predictors of MERS-CoV seropositivity. Our studies indicate a high level of seropositivity to MERS-CoV in camels in the counties surveyed, and highlights the important risk factors associated with MERS-CoV seropositivity in camels. Given that MERS-CoV is a zoonosis, and Kenya possesses the fourth largest camel population in Africa, these findings are important to inform the development of efficient and risk-based prevention and mitigation strategies against MERS-CoV transmission to humans.

Project description:BackgroundIn order to protect health workers from SARS-CoV-2, there is need to characterise the different types of patient facing health workers. Our first aim was to determine both the infection status and seroprevalence of SARS-CoV-2 in health workers. Our second aim was to evaluate the occupational and demographic predictors of seropositivity to inform the country's infection prevention and control (IPC) strategy.Methods and principal findingsWe invited 713 staff members at 24 out of 35 health facilities in the City of Bulawayo in Zimbabwe. Compliance to testing was defined as the willingness to uptake COVID-19 testing by answering a questionnaire and providing samples for both antibody testing and PCR testing. SARS-COV-2 antibodies were detected using a rapid diagnostic test kit and SAR-COV-2 infection was determined by real-time (RT)-PCR. Of the 713 participants, 635(89%) consented to answering the questionnaire and providing blood sample for antibody testing while 560 (78.5%) agreed to provide nasopharyngeal swabs for the PCR SARS-CoV-2 testing. Of the 635 people (aged 18-73) providing a blood sample 39.1% reported a history of past COVID-19 symptoms while 14.2% reported having current symptoms of COVID-19. The most-prevalent co-morbidity among this group was hypertension (22.0%) followed by asthma (7.0%) and diabetes (6.0%). The SARS-CoV-2 sero-prevalence was 8.9%. Of the 560 participants tested for SARS-CoV-2 infection, 2 participants (0.36%) were positive for SAR-CoV-2 infection by PCR testing. None of the SARS-CoV-2 antibody positive people were positive for SAR-CoV-2 infection by PCR testing.Conclusion and interpretationIn addition to clinical staff, several patient-facing health workers were characterised within Zimbabwe's health system and the seroprevalence data indicated that previous exposure to SAR-CoV-2 had occurred across the full spectrum of patient-facing staff with nurses and nurse aides having the highest seroprevalence. Our results highlight the need for including the various health workers in IPC strategies in health centres to ensure effective biosecurity and biosafety.

Dataset Information

Development and validation of different indirect ELISAs for MERS-CoV serological testing.

Publications

Development and validation of different indirect ELISAs for MERS-CoV serological testing.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets