A simple and high-throughput luciferase immunosorbent assay for both qualitative and semi-quantitative detection of anti-HIV-1 antibodies

Wang, Haiying; Cai, Qundi; Liang, Yuanhao; Shui, Jingwei; Tang, Shixing

doi:10.1016/j.virusres.2018.12.017

Cited by 15 publications

(19 citation statements)

References 19 publications

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“…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]. Cell lysates are directly used in the detection assay, which avoids purification and post-expression modifications [55].…”

Section: Discussionmentioning

confidence: 99%

“…LISA is characterized by rapid preparation, ultra-sensitivity, high-throughput, and simple operation [55,56]. Cell lysates are directly used in the detection assay, which avoids purification and post-expression modifications [55]. In the current study, we developed LISAs for the detection of anti-MERS-CoV IgG in human and animal sera based on novel and ultrasensitive NLuc approaches.…”

Section: Discussionmentioning

confidence: 99%

“…Compared to LIPS, LISA does not need pre-incubation of Ruc-antigen and samples and transfer of the mixture to a special 96-well filter plate containing the protein A/G beads. Moreover, LISA does not need a special plate washer with vacuum to capture the antigen-antibody-beads complex [55,68]. In the LISA system, Nluc replaced the Renilla luciferase used in LIPS.…”

Section: Discussionmentioning

confidence: 99%

“…An ultrasensitive and high-throughput luciferase immunosorbent assay (LISA) based on recombinant antigens fused with NanoLuc luciferase (NLuc) has been reported to detect IgG against several pathogens [55][56][57][58]. This assay has the advantage of not requiring a BSL-3 laboratory facility and species-specific labeled secondary antibodies for detection.…”

Section: Contents Lists Available At Sciencedirectmentioning

confidence: 99%

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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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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Contents Lists Available At Sciencedirectmentioning

confidence: 99%

See 2 more Smart Citations

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

Self Cite

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show abstract

“…The fusion protein was applied as a label in one-step competitive bioluminescent immunoassay for the detection of aflatoxin in cereal, which showed more than a 20-fold improvement in sensitivity as compared to classical ELISA [ 219 ]. NanoLuc fused with HIV-1 p24 or gp41 antigen was applied for qualitative and quantitative detection of anti-HIV-1 antibody in sera with the sensitivity exceeding that of ELISA 104-fold [ 220 ]. A similar assay was used to detect human norovirus-specific antibodies [ 221 ] or autoantibodies directed against proinsulin/insulin, which is important for diagnosis of type 1 diabetes [ 222 ].…”

Section: Ctz-dependent Luciferase Analytical Applicationmentioning

confidence: 99%

Coelenterazine-Dependent Luciferases as a Powerful Analytical Tool for Research and Biomedical Applications

Krasitskaya

Bashmakova

Frank

2020

IJMS

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: The functioning of bioluminescent systems in most of the known marine organisms is based on the oxidation reaction of the same substrate—coelenterazine (CTZ), catalyzed by luciferase. Despite the diversity in structures and the functioning mechanisms, these enzymes can be united into a common group called CTZ-dependent luciferases. Among these, there are two sharply different types of the system organization—Ca2+-regulated photoproteins and luciferases themselves that function in accordance with the classical enzyme–substrate kinetics. Along with deep and comprehensive fundamental research on these systems, approaches and methods of their practical use as highly sensitive reporters in analytics have been developed. The research aiming at the creation of artificial luciferases and synthetic CTZ analogues with new unique properties has led to the development of new experimental analytical methods based on them. The commercial availability of many ready-to-use assay systems based on CTZ-dependent luciferases is also important when choosing them by first-time-users. The development of analytical methods based on these bioluminescent systems is currently booming. The bioluminescent systems under consideration were successfully applied in various biological research areas, which confirms them to be a powerful analytical tool. In this review, we consider the main directions, results, and achievements in research involving these luciferases.

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Development and Evaluation of a Universal and Supersensitive NS1-Based Luciferase Immunosorbent Assay to Detect Zika Virus-Specific IgG

Wang

Zhan

et al. 2019

Virol. Sin.

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A simple and high-throughput luciferase immunosorbent assay for both qualitative and semi-quantitative detection of anti-HIV-1 antibodies

Cited by 15 publications

References 19 publications

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

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

Coelenterazine-Dependent Luciferases as a Powerful Analytical Tool for Research and Biomedical Applications

Development and Evaluation of a Universal and Supersensitive NS1-Based Luciferase Immunosorbent Assay to Detect Zika Virus-Specific IgG

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