Enzyme-linked immunosorbent assays using virus-like particles containing mutations of conserved residues on envelope protein can distinguish three flavivirus infections
The recent outbreak of Zika virus (ZIKV) in areas endemic flavivirus-highlight the need for sensitive and specific serological tests. Previously, we and others reported the fusion loop lock (FL) residue and / or the BC loop (BCL) residues on the envelope protein of dengue virus (DENV) is recognized by flavivirus cross-reactive human monoclonal antibodies and polyclonal sera.
To improve ZIKV serodiagnosis, we employ wild type (WT) and FL or FL / BCL mutant virus-like particle (VLP) from ZIKV, DENV1 and the West Nile virus (WNV) in enzyme linked immunosorbent assay (ELISA), and tested convalescent- phase serum or plasma samples from reverse-transcription PCR-confirmed cases with different ZIKV, DENV and WNV infection. For the IgG ELISA, ZIKV WT-VLP had a sensitivity of 100% and a specificity of 52.9%, which increased to 83.3% by FL / BCL mutant VLP and 92.2% with relative optical density ratio WT mutant VLP.
Similarly, DENV1 and WNV-VLP WT has a sensitivity / specificity of 100% / 70.0% and 100% / 56.3%, respectively; specificity increased to 93.3% and 83.0% by FL mutant VLP. For IgM ELISA, ZIKV, DENV1 and WNV-VLP WT has a specificity of 96.4%, 92.3% and 91.4%, respectively, for the primary infection; improved specificity of 93.7 to 99.3% by FL or FL / BCL mutant VLP. An algorithm based on a combination of mutant and WT-VLP IgG ELISA is proposed for the main ZIKV discrimination, DENV and WNV infection and secondary DENV infections and ZIKV with previous DENV infection; This can be a powerful tool to better understand the pathogenesis of seroprevalence and ZIKV in an area where several flaviviruses co-circulate.
serotype-specific detection of dengue virus nonstructural protein 1-based enzyme-linked immunosorbent assay validated by multi-national cohort
Background: Dengue virus (DENV) infection poses one the biggest obstacle to global human health. Four serotypes (DENV 1-4) present different symptoms and influence the immune responds next DENV infection, rendering surveillance, risk assessment, and control of diseases especially challenging. Early diagnosis and clinical management is very important and can be achieved by detection of DENV nonstructural protein 1 (NS1) in serum during the acute phase. However, some of the NS1-based test has been developed which is able to distinguish serotype DENV and none are currently available commercially.
Methodology / findings principles: We develop enzyme-linked immunosorbent assay (ELISA) to distinguish DENV-1-4 NS1 using serotype-specific pair of monoclonal antibodies. A total of 1,046 antibodies harvested from DENV-immunized mice and screened for antigen binding affinity. Clinical performance was evaluated using the ELISA-reaction confirmed 408 dengue polymerase chain samples obtained from patients in Brazil, Honduras, and India. The overall sensitivity of the test for the pan-DENV was 79.66% (325/408), and sensitivity to serotype DENV-1-4 were 79.1% (38/48), 80.41% (78/97), 100% ( 45/45) and 79.6% (98/123), respectively. Specificity reaches 94.07 to 100%.
Significance: Our research shows that strong antibody screening strategy that enables the development of ELISA-based NS1 serotype with a maximized specific and sensitive antigen binding. sensitive and specific assay was also used cohort most expansive to date, and where about half of Latin America, the geographic area is very under-represented in previous similar studies. ELISA test offers the potential for enhanced diagnosis during the acute phase of infection to help guide patient care and disease control.
Description: Quantitativesandwich ELISA kit for measuring Mouse diamine oxidase, DAO in samples from serum, plasma, tissue homogenates. Now available in a cost efficient pack of 5 plates of 96 wells each, conveniently packed along with the other reagents in 5 separate kits.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mouse Diamine Oxidase (DAO) in serum, plasma, tissue homogenates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mouse Diamine Oxidase (DAO) in serum, plasma, tissue homogenates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mouse Diamine Oxidase (DAO) in serum, plasma, tissue homogenates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mouse Diamine Oxidase (DAO) in serum, plasma, tissue homogenates and other biological fluids.
Description: Enzyme-linked immunosorbent assay based on the Double-antibody Sandwich method for detection of Mouse Diamine Oxidase (DAO) in samples from serum, plasma, tissue homogenates and other biological fluids with no significant corss-reactivity with analogues from other species.
Description: A sandwich ELISA kit for detection of Diamine Oxidase from Mouse in samples from blood, serum, plasma, cell culture fluid and other biological fluids.
Description: Quantitative sandwich ELISA for measuring Mouse Diamine oxidase (DAO) in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.
Description: Quantitative sandwich ELISA for measuring Mouse Diamine oxidase (DAO) in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.
Description: Quantitative sandwich ELISA for measuring Mouse Diamine oxidase (DAO) in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.
Description: Description of target: Regulates the level of the neuromodulator D-serine in the brain. Has high activity towards D-DOPA and contributes to dopamine synthesis. Could act as a detoxifying agent which removes D-amino acids accumulated during aging. Acts on a variety of D-amino acids with a preference for those having small hydrophobic side chains followed by those bearing polar, aromatic, and basic groups. Does not act on acidic amino acids.;Species reactivity: Rat;Application: ;Assay info: Assay Methodology: Quantitative Sandwich ELISA;Sensitivity: 0.195 mIU/mL
Description: Description of target: This gene encodes the peroxisomal enzyme D-amino acid oxidase. The enzyme is a flavoprotein which uses flavin adenine dinucleotide (FAD) as its prosthetic group. Its substrates include a wide variety of D-amino acids, but it is inactive on the naturally occurring L-amino acids. Its biological function is not known; it may act as a detoxifying agent which removes D-amino acids that accumulate during aging. In mice, it degrades D-serine, a co-agonist of the NMDA receptor. This gene may play a role in the pathophysiology of schizophrenia.;Species reactivity: Human;Application: ELISA;Assay info: Assay Methodology: Quantitative Sandwich ELISA;Sensitivity: 0.56 ng/mL
Description: Description of target: Regulates the level of the neuromodulator D-serine in the brain. Has high activity towards D-DOPA and contributes to dopamine synthesis. Could act as a detoxifying agent which removes D-amino acids accumulated during aging. Acts on a variety of D-amino acids with a preference for those having small hydrophobic side chains followed by those bearing polar, aromatic, and basic groups. Does not act on acidic amino acids.;Species reactivity: Rabbit;Application: ;Assay info: Assay Methodology: Quantitative Sandwich ELISA;Sensitivity: 0.312 U/mL
Description: Quantitativesandwich ELISA kit for measuring Rat diamine oxidase (DAO) in samples from serum, plasma, cell culture supernates, tissue homogenates. A new trial version of the kit, which allows you to test the kit in your application at a reasonable price.
Description: Quantitativesandwich ELISA kit for measuring Rat diamine oxidase(DAO) in samples from serum, plasma, cell culture supernates, tissue homogenates. Now available in a cost efficient pack of 5 plates of 96 wells each, conveniently packed along with the other reagents in 5 separate kits.
Description: A sandwich quantitative ELISA assay kit for detection of Rat Diamine Oxidase (DAO) in samples from serum, plasma, tissue homogenates, cell lysates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Rat Diamine Oxidase (DAO) in samples from serum, plasma, tissue homogenates, cell lysates or other biological fluids.
These results indicate that this ELISA is a promising aid in the early diagnosis of DENV-1-4 and supervision in areas of endemicity in addition to offering a comfortable monitoring for future vaccine interventions.
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