A novel luciferase immunosorbent assay performs better than a commercial enzyme-linked immunosorbent assay to detect MERS-CoV specific IgG in humans and animals
Middle East respiratory syndrome (mer) is a zoonosis caused by the mer deadly coronavirus (Mers-CoV) and pose a significant threat to public health worldwide. Therefore, serology rapid, sensitive, and specific for the detection of antibodies anti-Mers-CoV in both humans and animals is indispensable for the successful management of this disease. Here, we evaluated a wide range of novel luciferase immunosorbent assay (LISA) based on the nucleocapsid protein (NP) as well as fragments derived from the spike protein (S) including subunit 1 (S1), the N terminal domain (NTD), the domain of the receptor-binding (RBD) and subunit 2 (S2) from S to detect specific IgG Mers-CoV. fusion proteins, including nanoluciferase (NLuc) and various fragments derived from the NP or S protein of Mers-CoV, expressed in T cells are human embryonic kidney 293.
Lisas detectable anti-Mers-CoV IgG further developed using cell lysates expressing various fusion proteins. Panel samples of human or animal infected with Mers-CoV is used to analyze the sensitivity and specificity of various Lisas in reference to Mers-CoV RT-PCR, ELISA-based commercial S1 and particle pseudovirus neutralization test (ppNT). Our results indicate that S1, RBD-, and NP-Lisas more sensitive than the NTD- and S2-Lisas to detect anti-mer-CoV IgG.
Further, S1, RBD-, and NP-Lisas more sensitive (by at least 16-fold) compared with a commercially available ELISA S1. In addition, S1, RBD-, and NP-specifically recognized LISA anti-Mers-CoV IgG and do not cross-react with samples from other human CoV (OC43, 229E, HKU1, NL63) -infected patients. More importantly, this Lisas proven implementation and their reliability to detect anti-Mers-CoV IgG in the sample of camels, monkeys, and mice, among which the RBD-LISA exhibited excellent performance.
The results of this study indicate that Mers-CoV Novel RBD- and S1-Lisas is a very effective platform for rapid and sensitive detection of anti-mer-CoV IgG in human and animal samples. This test has the potential to be used as a serological test for the management and control of Mers-CoV infection.
A novel luciferase immunosorbent assay performs better than a commercial enzyme-linked immunosorbent assay to detect MERS-CoV specific IgG in humans and animals
Development of antigen-capture enzyme-linked immunosorbent assay and immunochromatographic strip based on monoclonal antibodies for the detection of the bird flu virus H6.
continuous surveillance has shown that the avian influenza virus subtype H6 (AIVs) which is prevalent in poultry and occasionally break barriers to infect the human species. Therefore it is necessary to build the specific, rapid and sensitive method to screen AIVs H6. In this study, a panel of monoclonal antibodies (mAbs) against hemagglutinin (HA) of the H6 AIV isolates produced.
Purified mAbs have high affinity and specificity to H6 AIVs. Antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) and immunochromatographic strip was developed based on two mAbs (1D7 and 1F12). Results of AC-ELISA shows high sensitivity with a limit of detection (LOD) of 3.9 ng / ml for HA protein and 0.5 H6 HAU (HA units) / 100 ml for life AIVs H6 subtypes. Average recovery AC-ELISA with allantoic fluid, respiratory specimens and cloacal swab was 91.907 ± 1.559%, 82.977% and 73.791 ± 1.497 ± 2.588%, respectively.
Description: Quantitative sandwich ELISA for measuring Human Creatinine (Cr) in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.
Description: A competitive inhibition quantitative ELISA assay kit for detection of Creatinine in samples from serum, plasma, tissue homogenates or other biological fluids.
Description: A competitive inhibition quantitative ELISA assay kit for detection of Creatinine in samples from serum, plasma, tissue homogenates or other biological fluids.
Description: A competitive ELISA for quantitative measurement of Rat Creatinine in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Rat Creatinine in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Rat Creatinine in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Porcine Creatinine in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Porcine Creatinine in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Porcine Creatinine in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Canine Creatinine in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Canine Creatinine in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Canine Creatinine in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with CTN protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to CTN. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of CTN in the samples is then determined by comparing the OD of the samples to the standard curve.
Description: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with CTN protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to CTN. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of CTN in the samples is then determined by comparing the OD of the samples to the standard curve.
The coefficient of intra and inter-assay variation of less than 10%. LOD immunochromatographic strip is 1 HAU when evaluated by the naked eye, and the detection time of less than 10 minutes without any equipment. Storage at room temperature or 4 ° C for 30 days or 60 days had no effect on the sensitivity and specificity of the strip. Thus, the AC-ELISA and immunochromatographic strip described here could be a secondary method for diagnosing infection AIV H6 with high specificity, sensitivity, and stability.
