scholarly journals Expansion and Refinement of Deep Sequence-Coupled Biopanning Technology for Epitope-Specific Antibody Responses in Human Serum

Viruses ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 1114
Author(s):  
Nikole L. Warner ◽  
Alexandria C. Linville ◽  
Susan B. Core ◽  
Brechla Moreno ◽  
Juan M. Pascale ◽  
...  
Keyword(s):  
Infectious Diseases ◽  
Deep Sequencing ◽  
Specific Antibody ◽  
Nonstructural Protein ◽  
Emerging Infectious Diseases ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Emerging Viruses ◽  
Nonstructural Protein 1 ◽  
Deep Sequencing Technology

Identifying the specific epitopes targeted by antibodies elicited in response to infectious diseases is important for developing vaccines and diagnostics. However, techniques for broadly exploring the specificity of antibodies in a rapid manner are lacking, limiting our ability to quickly respond to emerging viruses. We previously reported a technology that couples deep sequencing technology with a bacteriophage MS2 virus-like particle (VLP) peptide display platform for identifying pathogen-specific antibody responses. Here, we describe refinements that expand the number of patient samples that can be processed at one time, increasing the utility of this technology for rapidly responding to emerging infectious diseases. We used dengue virus (DENV) as a model system since much is already known about the antibody response. Sera from primary DENV-infected patients (n = 28) were used to pan an MS2 bacteriophage VLP library displaying all possible 10-amino-acid peptides from the DENV polypeptide. Selected VLPs were identified by deep sequencing and further investigated by enzyme-linked immunosorbent assay. We identified previously described immunodominant regions of envelope and nonstructural protein-1, as well as a number of other epitopes. Our refinement of the deep sequence-coupled biopanning technology expands the utility of this approach for rapidly investigating the specificity of antibody responses to infectious diseases.

Nanovaccine based on self-assembling nonstructural protein 1 boosts antibody responses to Zika virus

2021 ◽  
Vol 32 ◽  
pp. 102334
Author(s):  
Marianna Teixeira Pinho Favaro ◽  
Monica Josiane Rodrigues-Jesus ◽  
Alexia Adrianne Venceslau-Carvalho ◽  
Rúbens Prince Dos Santos Alves ◽  
Lennon Ramos Pereira ◽  
...  
Keyword(s):  
Zika Virus ◽  
Nonstructural Protein ◽  
Antibody Responses ◽  
Self Assembling ◽  
Nonstructural Protein 1

scholarly journals Use of a Rapid Test for Diagnosis of Dengue during Suspected Dengue Outbreaks in Resource-Limited Regions

2016 ◽  
Vol 54 (8) ◽  
pp. 2090-2095 ◽  
Author(s):  
Elizabeth A. Hunsperger ◽  
Tyler M. Sharp ◽  
Paul Lalita ◽  
Kini Tikomaidraubuta ◽  
Yolanda Rebello Cardoso ◽  
...  
Keyword(s):  
Public Health ◽  
Sensitivity And Specificity ◽  
Nonstructural Protein ◽  
Clinical Care ◽  
Rapid Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Major Public Health Problem ◽  
Public Health Response ◽  
Nonstructural Protein 1 ◽  
Dengue Outbreaks

Dengue is major public health problem, globally. Timely verification of suspected dengue outbreaks allows for public health response, leading to the initiation of appropriate clinical care. Because the clinical presentation of dengue is nonspecific, dengue diagnosis would benefit from a sensitive rapid diagnostic test (RDT). We evaluated the diagnostic performance of an RDT that detects dengue virus (DENV) nonstructural protein 1 (NS1) and anti-DENV IgM during suspected acute febrile illness (AFI) outbreaks in four countries. Real-time reverse transcription-PCR and anti-DENV IgM enzyme-linked immunosorbent assay were used to verify RDT results. Anti-DENV IgM RDT sensitivity and specificity ranged from 55.3 to 91.7% and 85.3 to 98.5%, respectively, and NS1 sensitivity and specificity ranged from 49.7 to 92.9% and 22.2 to 89.0%, respectively. Sensitivity varied by timing of specimen collection and DENV serotype. Combined test results moderately improved the sensitivity. The use of RDTs identified dengue as the cause of AFI outbreaks where reference diagnostic testing was limited or unavailable.

scholarly journals Co-circulation of all the Four Serotype of Dengue Virus in Endemic Region of Saurashtra, Gujarat during 2019-2020 Season

2021 ◽  
Author(s):  
Binita Joseph Aring ◽  
Dipali Magan Bhai Gavali ◽  
Pushpa Ramjibhai Kateshiya ◽  
Hiral Modbhai Gadhvi ◽  
Summaiya Mullan ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Nonstructural Protein ◽  
Geographical Area ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Endemic Region ◽  
Male Population ◽  
Monsoon Period ◽  
Cross Sectional ◽  
Nonstructural Protein 1

Introduction: Dengue has rapidly emerged as a vector - borne viral disease in recent years and also endemic in all continents. The agent of dengue, i.e., dengue viruses, are categorised under the genus Flavivirus, with the four dengue virus serotypes: designated as DENV-1, DENV-2, DENV-3 and DENV-4. These all four serotypes are in circulation either singly, or more than one at the same time. Aim: To study the epidemiological update of dengue with circulating serotype and co-infection in Saurashtra region, Gujarat, India. Materials and Methods: A cross-sectional study was conducted during January 2019 to December 2020 and total samples received were 12,563 which were clinically suspected dengue samples case. After receiving blood samples, serum was separated and proceeded for Dengue NS1Ag (nonstructural protein 1 antigen), and Dengue IgM Ab (Immunoglobulin M antibody). After serological confirmation, 151 samples from different geographical area were selected for Dengue specific Reverse Transcriptase- Polymerase Chain Reaction (RT-PCR) for serotyping. results: Total 4069 (32%) had confirmed dengue positive by Enzyme Linked Immunosorbent Assay (ELISA). The ratio of male cases was higher than female, and in age group 21-35 year (47%). Seasonal trend showed a gradual increase in positivity from June with high peak in October. Circulation of all the four serotypes in area, higher monotypic infection by DENV-1 serotype (41%), followed by DENV-4, DENV-2 and DENV-3. Co-infection of different serotypes were also found. conclusion: The present study concluded that all four serotypes circulate with predominant being DENV-1 type and co-infection of different serotypes in the saurashtra region. Dengue mainly affected adult male population, and seasonal peak during monsoon and post-monsoon period.

scholarly journals Limitations of Pneumovax® as a detection antigen in the measurement of serotype-specific antibodies by enzyme-linked immunosorbent assay

2002 ◽  
Vol 39 (4) ◽  
pp. 398-403 ◽  
Author(s):  
ZM Huo ◽  
J Miles ◽  
PG Riches ◽  
T Harris
Keyword(s):  
Immunosorbent Assay ◽  
Specific Antibody ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Carbohydrate Antigens ◽  
Specific Antibodies ◽  
Elisa System ◽  
Background Measurement ◽  
Polysaccharide Antigens ◽  
The Individual

Background: Measurement of antibody responses to polysaccharide antigens is regarded as an important assessment of an individual's ability to respond to carbohydrate antigens. The currently used assays for the measurement of pneumococcal-specific antibody use the multi-serotype vaccine Pneumovax® as the detection antigen. Methods: An equal potency enzyme-linked immunosorbent assay (ELISA) system was used to compare the measurement of serotype-specific antibody with the multi-serotype assay. Results: Our results show that the concentration of specific antibody to Pneumovax is not related to the concentration of antibody to the individual serotypes. Neither is any correlation found between the antibody concentrations to any of the three single serotypes investigated, to the mixture of the three serotypes or to Pneumovax. Conclusion: We conclude that the measurement of the concentration of the specific antibody to the mixed serotypes present in Pneumovax has serious limitations when used to evaluate the protection acquired from Pneumovax immunization against any specific serotype.

scholarly journals Detection of Pseudorabies Virus Infection in Subunit-Vaccinated and Nonvaccinated Pigs using a Nucleocapsid-Based Enzyme-Linked Immunosorbent Assay

1992 ◽  
Vol 4 (2) ◽  
pp. 164-169 ◽  
Author(s):  
M. J. McGinley ◽  
D. L. Todd ◽  
H. T. Hill ◽  
K. B. Platt
Keyword(s):  
Immunosorbent Assay ◽  
Specific Antibody ◽  
Nucleocapsid Protein ◽  
Pseudorabies Virus ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Screening Assay ◽  
Virus Specific Antibody ◽  
Positive Threshold ◽  
Virus Exposure

The potential of a pseudorabies virus (PRV) nucleocapsid protein (NC)-based enzyme-linked immunosorbent assay (ELISA) as a screening assay for PRV infection in subunit-vaccinated and nonvaccinated pigs was studied. The NC-ELISA compared favorably to a commercial ELISA for detecting PRV infection in nonvaccinated pigs. Virus-specific antibody was first detected by the NC-ELISA between days 14 and 21 in 5 pigs challenged intranasally with 104PFU of virus. Antibody continued to be detected in these pigs through day 42, when the experiment was terminated. The NC-ELISA also detected antibody in 23 of 24 pigs from PRV-infected herds. In contrast, the commercial ELISA detected antibody 1 week earlier than the NC-ELISA in experimentally infected pigs but failed to detect antibody in 3 naturally exposed pigs that were identified by the NC-ELISA. Infection in these animals was confirmed by radioimmunoprecipitation analysis. The potential usefulness of the NC-ELISA for detecting infection in vaccinated pigs was also evaluated. The nucleocapsid-specific antibody responses of 10 PRV envelope glycoprotein subunit-vaccinated pigs were monitored prior to and following nasal exposure to a low dose (1023PFU) of PRV. Sera were collected periodically for 113 days after infection. Nucleocapsid-specific antibody responses measured by the NC-ELISA remained below the positive threshold before challenge but increased dramatically following virus exposure. Maximum ELISA responses were obtained on day 32 postchallenge (p.c.). Mean ELISA responses decreased thereafter but remained well above the positive threshold on day 113 p.c. PRV nucleocapsid protein can be used effectively as antigen in the ELISA for detecting PRV infection in both nonvaccinated and subunit-vaccinated pigs.

scholarly journals Antibody Responses in Reindeer (Rangifer tarandus) Infected with Mycobacterium bovis

2005 ◽  
Vol 12 (6) ◽  
pp. 727-735 ◽  
Author(s):  
W. R. Waters ◽  
M. V. Palmer ◽  
J. P. Bannantine ◽  
R. Greenwald ◽  
J. Esfandiari ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Specific Antibody ◽  
Rangifer Tarandus ◽  
Skin Testing ◽  
Serum Antibody ◽  
The United States ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cell Mediated Immunity ◽  
Testing Procedures

ABSTRACT Despite having a very low incidence of disease, reindeer (Rangifer tarandus) are subject to tuberculosis (TB) testing requirements for interstate shipment and herd accreditation in the United States. Improved TB tests are desperately needed, as many reindeer are falsely classified as reactors by current testing procedures. Sera collected sequentially from 11 (experimentally) Mycobacterium bovis-infected reindeer and 4 noninfected reindeer were evaluated by enzyme-linked immunosorbent assay (ELISA), immunoblotting, and multiantigen print immunoassay (MAPIA) for antibody specific to M. bovis antigens. Specific antibody was detected as early as 4 weeks after challenge with M. bovis. By MAPIA, sera were tested with 12 native and recombinant antigens, which were used to coat nitrocellulose. All M. bovis-infected reindeer developed responses to MPB83 and a fusion protein, Acr1/MPB83, and 9/11 had responses to MPB70. Other antigens less commonly recognized included MPB59, ESAT-6, and CFP10. Administration of purified protein derivatives for skin testing boosted serum antibody responses, as detected by each of the assays. Of the noninfected reindeer, 2/4 had responses that were detectable immediately following skin testing, which correlated with pathological findings (i.e., presence of granulomatous lesions yet the absence of acid-fast bacteria). The levels of specific antibody produced by infected reindeer appeared to be associated with disease progression but not with cell-mediated immunity. These findings indicate that M. bovis infection of reindeer elicits an antibody response to multiple antigens that can be boosted by skin testing. Serological tests using carefully selected specific antigens have potential for early detection of infections in reindeer.

scholarly journals Circulation of DENV-3 Genotype 3 during 2017 to 2018 in Delhi: A Single-Center Hospital-Based Study

2021 ◽  
Author(s):  
Abhishek Padhi ◽  
Ekta Gupta ◽  
Gaurav Singh ◽  
Shama Parveen ◽  
Arshi Islam ◽  
...  
Keyword(s):  
Acute Phase ◽  
Nonstructural Protein ◽  
Indian Subcontinent ◽  
Enzyme Linked Immunosorbent Assay ◽  
Care Hospital ◽  
Rt Pcr ◽  
Female Ratio ◽  
Genotype Iii ◽  
Nonstructural Protein 1 ◽  
New Variant

Abstract Introduction Delhi is hyperendemic for dengue virus (DENV) where all the four DENV have previously been reported. A constant vigilance of circulating DENV serotypes is important in surveillance, since the introduction of a new variant to areas affected by preexisting serotypes constitutes a risk factor for dengue hemorrhagic fever and dengue shock syndrome. Objectives This retrospective study was performed with an objective to determine the circulating serotype and genotype of DENV in acute phase blood samples of patients who have reported to a tertiary liver care hospital in New Delhi during the last 2 years (2017–2018). Methods The data of clinician-initiated testing for dengue nonstructural protein 1 (NS1) antigen (Ag) was searched in the institutional hospital information system. The serum sample of dengue NS1 Ag-positive cases confirmed by enzyme-linked immunosorbent assay (ELISA; PANBIO, Gyeonggi-do, ROK) and a fever duration of less than 5 days were retrieved from the laboratory archive. The DENV serotyping on these sample was performed by reverse transcriptase polymerase chain reaction (RT-PCR). Sequencing and phylogenetic analysis was done for the capsid premembrane (CprM) region to determine the genotype. Results A total of 440 acute-phase samples were received. Twenty one (4.77%) were positive for dengue NS1 Ag with a mean age of 35.1 years and male-to-female ratio of 1.1:1. Eight cases (38.09%) were positive by dengue RT-PCR and all belonged to DENV-3 serotypes. Phylogenetic tree analysis revealed DENV-3 clustered to genotype III with 100% homology with 2008 Indian subcontinent strain. Conclusion This study revealed circulation of DENV-3, genotype III in Delhi from 2017 to 2018, similar to the 2008 viral type. Virological surveillance is an important exercise to be done for viral infections with public threat and outbreak potential.

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