Suggested new breakpoints of anti-MERS-CoV antibody ELISA titers: performance analysis of serologic tests

Ko, Jae-Hoon; Müller, Marcel A.; Seok, Hyeri; Park, G. E.; Lee, J. Y.; Cho, S. Y.; Ha, Young Eun; Baek, Jin Yang; Kim, S. H.; Kang, Jeung-Mo; Kim, Y.-J.; Jo, Ik Joon; Chung, Chi Ryang; Hahn, Myong‐Joon; Drosten, Christian; Kang, Cheol-In; Chung, Doo Ryeon; Song, Jae-Hoon; Kang, Eun‐Suk; Peck, Kyong Ran

doi:10.1007/s10096-017-3043-3

Cited by 22 publications

(22 citation statements)

References 15 publications

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“…Recent work examining a panel of MERS‐CoV patient serum has suggested that S ELISA sensitivity can be increased by lowering the breakpoint in MERS‐CoV S ELISAs . While doing this would capture a larger percentage of patients who seroconvert, we have seen several patients who never develop detectable MERS‐CoV S antibodies, but do develop N antibodies (data not shown).…”

Section: Resultsmentioning

confidence: 99%

Inclusion of MERS‐spike protein ELISA in algorithm to determine serologic evidence of MERS‐CoV infection

Trivedi

Miao

Alabdallat

et al. 2017

Journal of Medical Virology

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The Centers for Disease Control and Prevention (CDC) algorithm for detecting presence of serum antibodies against Middle East Respiratory Syndrome coronavirus (MERS-CoV) in subjects with potential infections with the virus has included screening by indirect ELISA against recombinant nucleocapsid (N) protein and confirmation by immunofluorescent staining of infected monolayers and/or micro-neutralization titration. Other international groups include indirect ELISA assays using the spike (S) protein, as part of their serological determinations. In the current study, we describe development and validation of an indirect MERS-CoV S ELISA to be used as part of our serological determination for evidence of previous exposure to the virus.

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Section: Resultsmentioning

confidence: 99%

Inclusion of MERS‐spike protein ELISA in algorithm to determine serologic evidence of MERS‐CoV infection

Trivedi

Miao

Alabdallat

et al. 2017

Journal of Medical Virology

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“…The present study used EUROIMMUN ELISA based on the purified spike protein domain S1 antigen for determining the seropositivity rate in camels and humans. The performance of this kit for testing MERS-CoV infection has been previously evaluated and it was stated that using a confirmatory test after initial screening of samples with this ELISA is not mandatory [33]. It has been also successfully used by other authors with 99% specificity compared to microneutalization test [11].…”

Section: Discussionmentioning

confidence: 99%

“…Semi-quantitative anti-MERS-CoV IgG camel and human ELISAs (Euroimmun, Lübeck, Germany) were used in the present study to detect specific IgG antibodies against MERS-CoV in camel and human sera, respectively. The kits utilize the purified spike protein domain S1 antigen of MERS coronavirus (MERS-CoV S1) based on its high sensitivity and high specificity [11,13,33]. Briefly, the serum samples were diluted (1:101) and the test procedure was then performed as per the manufacturer's instructions.…”

Section: Serological Assaymentioning

confidence: 99%

Seroprevalence of Middle East Respiratory Syndrome Corona Virus in dromedaries and their traders in upper Egypt

Sayed

Malek

Abushahba

2020

J Infect Dev Ctries

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Introduction: Camel trade in Egypt depends mainly on importation. Seemingly healthy imported camels are responsible for the ingress of serious diseases into Egypt. A striking example of this concerning public health globally is the Middle East respiratory coronavirus (MERS-CoV) which causes case fatalities of over 34%. Here, we determined the seroepidemiological situation of the MERS-CoV in imported camels and their traders in Upper Egypt. Methodology: Sera of sixty-three dromedaries and twenty-eight camel traders were recruited (January 2015-December 2016). The age, gender, and sampling locality of each sampled camel and human were obtained. Semi-quantitative anti-MERS-CoV IgG ELISAs which utilize the purified spike protein domain S1 antigen of MERS coronavirus (MERS-CoV S1) were used to detect specific IgG antibodies against the virus. Results: The data showed that 58.73% of imported camels and 25% of traders had antibodies specific to MERS-CoV. Interestingly, like seroreactive camels, all seropositive humans were apparently healthy without any history of developing severe respiratory disease in the 14 days prior to sampling. Having specific antibodies among the examined camel sera was significantly different (P < 0.0001) in relation to various sampling localities, gender and age groups. In contrast, the seropositivity rate of MERS-CoV IgG in humans did not differ significantly by any of the studied factors. Conclusions: The current study provides the first serological evidence of occupational exposure of humans to MERS-CoV in Africa. Additionally, it reports that imported camels could be implicated in introducing MERS-CoV into Egypt. Accordingly, application of strict control measures to camel importation is a priority.

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“…However, these assays are timeconsuming, laborious, and require high-level biosafety facilities [53,60]. The performance of the commercial MERS-CoV S1-based ELISA has been assessed by different groups [18,35,51,65,66], all of which have shown that the sensitivity of this assay needs to be improved, as it shows high cut-off values from the optical density (OD) ratio (the manufacturer's instructions provided a higher breakpoint for ELISA IgG to warrant specificity, with a borderline OD ratio cut-off of 0.8-1.1 and positive ≥1.1). LISA is characterized by rapid preparation, ultra-sensitivity, high-throughput, and simple operation [55,56].…”

Section: Discussionmentioning

confidence: 99%

“…Several methods can be used to detect serum antibodies against MERS-CoV, such as enzyme-linked immunosorbent assay (ELISA), microneutralization (MN), immunofluorescence assay (IFA), the plaque reduction neutralization test (PRNT), and the pseudovirus particle neutralization test (ppNT). Among these assays, MN, IFA, and PRNT depend on virus cultivation, and thus, require equipment and facilities located within a Biosafety Level 3 (BSL-3) laboratory [1,3,12,18,35,39,[50][51][52][53][54]. While there are commercial anti-MERS-CoV serologic test kits for humans and animals including ELISA and IFA, rapid and sensitive serological assays for the detection of anti-MERS-CoV IgG independent of purified protein, specific secondary antibody, virus cultivation and BSL-3 facilities are still urgently needed.…”

Section: Contents Lists Available At Sciencedirectmentioning

confidence: 99%

A novel luciferase immunosorbent assay performs better than a commercial enzyme-linked immunosorbent assay to detect MERS-CoV specific IgG in humans and animals

Wang¹,

Wang²,

Deng³

et al. 2019

Biosafety and Health

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The Middle East respiratory syndrome (MERS) is a lethal zoonosis caused by MERS coronavirus (MERS-CoV) and poses a significant threat to public health worldwide. Therefore, a rapid, sensitive, and specific serologic test for detecting anti-MERS-CoV antibodies in both humans and animals is urgently needed for the successful management of this illness. Here, we evaluated various novel luciferase immunosorbent assays (LISA) based on nucleocapsid protein (NP) as well as fragments derived from spike protein (S) including subunit 1 (S1), N terminal domain (NTD), receptorbinding domain (RBD) and subunit 2 (S2) of S for the detection of MERS-CoV-specific IgG. Fusion proteins, including nanoluciferase (NLuc) and various fragments derived from the NP or S protein of MERS-CoV, were expressed in human embryonic kidney 293 T cells. LISAs that detected anti-MERS-CoV IgG were further developed using cell lysates expressing various fusion proteins. Panels of human or animal samples infected with MERS-CoV were used to analyze the sensitivity and specificity of various LISAs in reference to a MERS-CoV RT-PCR, commercial S1-based ELISA, and pseudovirus particle neutralization test (ppNT). Our results showed that the S1-, RBD-, and NP-LISAs were more sensitive than the NTD-and S2-LISAs for the detection of anti-MERS-CoV IgG. Furthermore, the S1-, RBD-, and NP-LISAs were more sensitive (by at least 16-fold) than the commercially available S1-ELISA. Moreover, the S1-, RBD-, and NP-LISA specifically recognized anti-MERS-CoV IgG and did not cross-react with samples derived from other human CoV (OC43, 229E, HKU1, NL63)-infected patients. More importantly, these LISAs proved their applicability and reliability for detecting anti-MERS-CoV IgG in samples from camels, monkeys, and mice, among which the RBD-LISA exhibited excellent performance. The results of this study suggest that the novel MERS-CoV RBD-and S1-LISAs are highly effective platforms for the rapid and sensitive detection of anti-MERS-CoV IgG in human and animal samples. These assays have the potential to be used as serologic tests for the management and control of MERS-CoV infection.

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Suggested new breakpoints of anti-MERS-CoV antibody ELISA titers: performance analysis of serologic tests

Cited by 22 publications

References 15 publications

Inclusion of MERS‐spike protein ELISA in algorithm to determine serologic evidence of MERS‐CoV infection

Inclusion of MERS‐spike protein ELISA in algorithm to determine serologic evidence of MERS‐CoV infection

Seroprevalence of Middle East Respiratory Syndrome Corona Virus in dromedaries and their traders in upper Egypt

A novel luciferase immunosorbent assay performs better than a commercial enzyme-linked immunosorbent assay to detect MERS-CoV specific IgG in humans and animals

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