scholarly journals Development of a Colloidal Gold Immunochromatographic Assay for Duck Enteritis Virus Detection Using Monoclonal Antibodies

Pathogens ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 365
Author(s):  
Fengli Liu ◽  
Yanxin Cao ◽  
Maokai Yan ◽  
Mengxu Sun ◽  
Qingshui Zhang ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Colloidal Gold ◽  
Preventive Intervention ◽  
Virus Detection ◽  
Cross Reactivity ◽  
Duck Enteritis Virus ◽  
Detection Sensitivity ◽  
Immunochromatographic Assay ◽  
Efficient Detection ◽  
Enteritis Virus

Duck viral enteritis is a highly contagious and fatal disease of commercial waterfowl flocks. The disease occurs sporadically or epizootically in mainland China due to insufficient vaccinations. Early and rapid diagnosis is important for preventive intervention and the control of epizootic events in clinical settings. In this study, we generated two monoclonal antibodies (MAbs) that specifically recognized the duck enteritis virus (DEV) envelope glycoprotein B and tegument protein UL47, respectively. Using these MAbs, a colloidal gold-based immunochromatographic assay (ICA) was developed for the efficient detection of DEV antigens within 15 min. Our results showed that the detection limit of the developed ICA strip was 2.52 × 103 TCID50/mL for the virus infected cell culture suspension with no cross-reactivity with other pathogenic viruses commonly encountered in commercially raised waterfowl. Using samples from experimentally infected ducks, we demonstrated that the ICA detected the virus in cloacal swab samples on day three post-infection, demonstrating an 80% concordance with the PCR. For tissue homogenates from ducks succumbing to infection, the detection sensitivity was 100%. The efficient and specific detection by this ICA test provides a valuable, convenient, easy to use and rapid diagnostic tool for DVE under both laboratory and field conditions.

scholarly journals Development of Monoclonal Antibodies against HIV-1 p24 Protein and Its Application in Colloidal Gold Immunochromatographic Assay for HIV-1 Detection

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
Yi Ma ◽  
Chao Ni ◽  
Emmanuel E. Dzakah ◽  
Haiying Wang ◽  
Keren Kang ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Early Diagnosis ◽  
Colloidal Gold ◽  
Limit Of Detection ◽  
Sandwich Elisa ◽  
Immunochromatographic Assay ◽  
Suitable Alternative ◽  
P24 Protein ◽  
Hybridoma Cell Lines ◽  
Hiv 1

Human immunodeficiency virus type 1 (HIV-1) p24 protein is the most abundant viral protein of HIV-1. This protein is secreted in blood serum at high levels during the early stages of HIV-1 infection, making it a biomarker for early diagnosis. In this study, a colloidal gold immunochromatographic assay (GICA) was established for detecting p24 protein using mouse monoclonal antibodies (mAbs). The HIV-1 p24 protein was expressed inE. colistrain BL21 and the purified protein was used to immunize mice. Stable hybridoma cell lines secreting anti-p24 monoclonal antibodies were obtained after ELISA screening and subcloning by limiting dilution. 34 different capture and labeling mAb pairs were selected by a novel antibody-capture indirect sandwich ELISA and then applied in GICA to detect p24 protein. The GICA method has a limit of detection (LOD) of 25 pg/mL and could detect p24 protein in all 10 positive samples obtained from the National Reference of HIV-1 p24 antigen. Out of 153 negative samples tested, 3 false positives results were obtained. The overall specificity of this test was 98.03%. The good sensitivity and specificity of this method make it a suitable alternative to provide a more convenient and efficient tool for early diagnosis of HIV infection.

Development of a Colloidal Gold-Based Immunochromatographic Assay for the Rapid Detection of Aflatoxin B1 in Food Samples

2013 ◽  
Vol 726-731 ◽  
pp. 1279-1282 ◽  
Author(s):  
Xue Qing Chen ◽  
Sha Sha Lu ◽  
Qing Sun ◽  
Jing Chen Yang ◽  
Guo Qing Shi
Keyword(s):  
Colloidal Gold ◽  
Rapid Detection ◽  
Limit Of Detection ◽  
Food Samples ◽  
Cross Reactivity ◽  
Immunochromatographic Assay ◽  
Rapid Testing ◽  
Elisa Method ◽  
Standard Solution ◽  
Aflatoxin B

A rapid and sensitive competitive immunochromatographic assay for aflatoxin B1 (AFB1) based on colloidal gold-antibody probe was developed. The limit of detection for AFB1 standard solution was 0.5ng/ml and for foodstuff sample was 10μg/Kg. Other aflatoxins including AFM1, AFB2, AFG1, and AFG2 have less cross-reactivity to the strip. The storge life of the strip was at least 12 monthes. The method was well agreement with the ELISA method by testing food samples. This method is suitable for rapid testing of AFB1 on site.

scholarly journals A New Workflow to Generate Monoclonal Antibodies against Microorganisms

2021 ◽  
Vol 11 (20) ◽  
pp. 9359
Author(s):  
Markus Göthel ◽  
Martin Listek ◽  
Katrin Messerschmidt ◽  
Anja Schlör ◽  
Anja Hönow ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Legionella Pneumophila ◽  
Initial Study ◽  
Cross Reactivity ◽  
Cell Surface Receptor ◽  
Peptide Epitopes ◽  
E Coli ◽  
Efficient Detection ◽  
Surface Receptor ◽  
Fusion Cell

Monoclonal antibodies are used worldwide as highly potent and efficient detection reagents for research and diagnostic applications. Nevertheless, the specific targeting of complex antigens such as whole microorganisms remains a challenge. To provide a comprehensive workflow, we combined bioinformatic analyses with novel immunization and selection tools to design monoclonal antibodies for the detection of whole microorganisms. In our initial study, we used the human pathogenic strain E. coli O157:H7 as a model target and identified 53 potential protein candidates by using reverse vaccinology methodology. Five different peptide epitopes were selected for immunization using epitope-engineered viral proteins. The identification of antibody-producing hybridomas was performed by using a novel screening technology based on transgenic fusion cell lines. Using an artificial cell surface receptor expressed by all hybridomas, the desired antigen-specific cells can be sorted fast and efficiently out of the fusion cell pool. Selected antibody candidates were characterized and showed strong binding to the target strain E. coli O157:H7 with minor or no cross-reactivity to other relevant microorganisms such as Legionella pneumophila and Bacillus ssp. This approach could be useful as a highly efficient workflow for the generation of antibodies against microorganisms.

Proteomic analysis of host cellular proteins co-immunoprecipitated with duck enteritis virus gC

2021 ◽  
Vol 245 ◽  
pp. 104281
Author(s):  
Liu Chen ◽  
Zheng Ni ◽  
Jionggang Hua ◽  
Weicheng Ye ◽  
Keshu Liu ◽  
...  
Keyword(s):  
Proteomic Analysis ◽  
Duck Enteritis Virus ◽  
Cellular Proteins ◽  
Enteritis Virus

Development of an antigen-ELISA and a colloidal gold–based immunochromatographic strip based on monoclonal antibodies for detection of avian influenza A(H5) viruses

2021 ◽  
pp. 104063872110275
Author(s):  
Yixin Xiao ◽  
Fan Yang ◽  
Fumin Liu ◽  
Linfang Cheng ◽  
Hangping Yao ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Disease Virus ◽  
Colloidal Gold ◽  
Rapid Detection ◽  
Influenza A ◽  
Field Investigation ◽  
Cross Reactivity ◽  
Detection Methods ◽  
Immunochromatographic Strip ◽  
Antigen Elisa

Avian influenza A(H5) viruses (avian IAVs) pose a major threat to the economy and public health. We developed an antigen-ELISA (ag-ELISA) and a colloidal gold–based immunochromatographic strip for the rapid detection of avian A(H5) viruses. Both detection methods displayed no cross-reactivity with other viruses (e.g., other avian IAVs, infectious bursal disease virus, Newcastle disease virus, infectious bronchitis virus, avian paramyxovirus). The ag-ELISA was sensitive down to 0.5 hemagglutinin (HA) units/100 µL of avian A(H5) viruses and 7.5 ng/mL of purified H5 HA proteins. The immunochromatographic strip was sensitive down to 1 HA unit/100 µL of avian A(H5) viruses. Both detection methods exhibited good reproducibility with CVs < 10%. For 200 random poultry samples, the sensitivity and specificity of the ag-ELISA were 92.6% and 98.8%, respectively, and for test strips were 88.9% and 98.3%, respectively. Both detection methods displayed high specificity, sensitivity, and stability, making them suitable for rapid detection and field investigation of avian A(H5) viruses.

scholarly journals Monoclonal antibodies to bovine UDP-galactosyltransferase. Characterization, cross-reactivity, and utilization as structural probes.

1986 ◽  
Vol 261 (17) ◽  
pp. 7975-7981
Author(s):  
J T Ulrich ◽  
J R Schenck ◽  
H G Rittenhouse ◽  
N L Shaper ◽  
J H Shaper
Keyword(s):  
Monoclonal Antibodies ◽  
Cross Reactivity ◽  
Structural Probes

scholarly journals Recombinase-Aided Amplification Coupled with Lateral Flow Dipstick for Efficient and Accurate Detection of Porcine Parvovirus

Life ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 762
Author(s):  
Yihong He ◽  
Wenxian Chen ◽  
Jindai Fan ◽  
Shuangqi Fan ◽  
Hongxing Ding ◽  
...  
Keyword(s):  
Detection Method ◽  
Cross Reactivity ◽  
Lateral Flow ◽  
Porcine Parvovirus ◽  
Swine Industry ◽  
Resource Limited ◽  
Efficient Detection ◽  
Lateral Flow Dipstick ◽  
Polymerase Chain ◽  
Embryonic Death

Porcine parvovirus (PPV) infection is the primary cause of SMEDI (stillbirth; mummification; embryonic death; infertility) syndrome, which is a global burden for the swine industry. Thus, it is crucial to establish a rapid and efficient detection method against PPV infection. In the present work, we developed a recombinase-aided amplification (RAA) assay, coupled with a lateral flow dipstick (LFD), to achieve an amplification of PPV DNA at 37 °C within 15 min. The detection limits of PPV RAA-LFD assay were 102 copies/μL recombinant plasmid pMD19-T-VP1, 6.38 × 10−7 ng/μL PPV DNA, and 10−1 TCID50/mL virus, respectively. This method was highly specific for PPV detection with no cross-reactivity for other swine pathogens. In contrast to polymerase chain reaction (PCR), the PPV RAA-LFD assay is more sensitive and cost-saving. Hence, the established PPV RAA-LFD assay provided an alternative for PPV detection, especially in resource-limited regions.

scholarly journals Screening and development of monoclonal antibodies for identification of ferret T follicular helper cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Wenbo Jiang ◽  
Julius Wong ◽  
Hyon-Xhi Tan ◽  
Hannah G. Kelly ◽  
Paul G. Whitney ◽  
...  
Keyword(s):  
Animal Model ◽  
Monoclonal Antibodies ◽  
Lymph Node ◽  
Cross Reactivity ◽  
Tfh Cells ◽  
Follicular Helper ◽  
Lymph Node Cells ◽  
Human Viruses ◽  
Humoral Responses ◽  
Murine Monoclonal Antibodies

AbstractThe ferret is a key animal model for investigating the pathogenicity and transmissibility of important human viruses, and for the pre‐clinical assessment of vaccines. However, relatively little is known about the ferret immune system, due in part to a paucity of ferret‐reactive reagents. In particular, T follicular helper (Tfh) cells are critical in the generation of effective humoral responses in humans, mice and other animal models but to date it has not been possible to identify Tfh in ferrets. Here, we describe the screening and development of ferret-reactive BCL6, CXCR5 and PD-1 monoclonal antibodies. We found two commercial anti-BCL6 antibodies (clone K112-91 and clone IG191E/A8) had cross-reactivity with lymph node cells from influenza-infected ferrets. We next developed two murine monoclonal antibodies against ferret CXCR5 (clone feX5-C05) and PD-1 (clone fePD-CL1) using a single B cell PCR-based method. We were able to clearly identify Tfh cells in lymph nodes from influenza infected ferrets using these antibodies. The development of ferret Tfh marker antibodies and the identification of ferret Tfh cells will assist the evaluation of vaccine-induced Tfh responses in the ferret model and the design of novel vaccines against the infection of influenza and other viruses, including SARS-CoV2.

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