scholarly journals A Novel Method of Safely Measuring Influenza Virus Aerosol Using Antigen-Capture Enzyme-Linked Immunosorbent Assay for the Performance Evaluation of Protective Clothing Materials

2016 ◽  
Vol 21 (2) ◽  
pp. 81-89 ◽  
Author(s):  
NORIKO SHIMASAKI ◽  
YASUHIRO NOJIMA ◽  
AKIRA OKAUE ◽  
HITOSHI TAKAHASHI ◽  
TSUTOMU KAGEYAMA ◽  
...  
2007 ◽  
Vol 14 (5) ◽  
pp. 617-623 ◽  
Author(s):  
Qigai He ◽  
Sumathy Velumani ◽  
Qingyun Du ◽  
Chee Wee Lim ◽  
Fook Kheong Ng ◽  
...  

ABSTRACT The unprecedented spread of highly pathogenic avian influenza virus subtype H5N1 in Asia and Europe is threatening animals and public health systems. Effective diagnosis and control management are needed to control the disease. To this end, we developed a panel of monoclonal antibodies (MAbs) against the H5N1 avian influenza virus (AIV) and implemented an antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) to detect the H5 viral antigen. Mice immunized with denatured hemagglutinin (HA) from A/goose/Guangdong/97 (H5N1) expressed in bacteria or immunized with concentrated H5N2 virus yielded a panel of hybridomas secreting MAbs specific for influenza virus HA. The reactivity of each MAb with several subtypes of influenza virus revealed that hybridomas 3D4 and 8B6 specifically recognized H5 HA. Therefore, purified antibodies from hybridomas 3D4 and 8B6, which secrete immunoglobulin G (IgG) and IgM, respectively, were used as the capture antibodies and pooled hyperimmune guinea pig serum IgG served as the detector antibody. The specificity of the optimized AC-ELISA was evaluated by using AIV subtypes H5 H3, H4, H7, H9, and H10. Specimens containing AIV subtype H5 subtype yielded a specific and strong signal above the background, whereas specimens containing all other subtypes yielded background signals. The detection limits of the AC-ELISA were 62.5 ng of bacterium-expressed H5N1 HA1 protein and 124, 62, and 31 50% tissue culture infective doses of influenza virus subtypes H5N1/PR8, H5N2, and H5N3, respectively. Reconstituted clinical samples consisting of H5 AIVs mixed with pharyngeal-tracheal mucus from healthy chickens also yielded positive signals in the AC-ELISA, and the results were confirmed by reverse transcription-PCR. The tracheal swab samples from H9N2-infected chickens did not give positive signals. Taken together, the newly developed MAb-based AC-ELISA offers an attractive alternative to other diagnostic approaches for the specific detection of H5 AIV.


2017 ◽  
Vol 48 (3) ◽  
pp. 882-885 ◽  
Author(s):  
Trine Hammer Jensen ◽  
Jannie Holmegaard Andersen ◽  
Charlotte Kristiane Hjulsager ◽  
Mariann Chriél ◽  
Mads Frost Bertelsen

2014 ◽  
Vol 21 (9) ◽  
pp. 1364-1368 ◽  
Author(s):  
Diego H. Caceres ◽  
Christina M. Scheel ◽  
Ángela M. Tobón ◽  
Angela Ahlquist Cleveland ◽  
Ángela Restrepo ◽  
...  

ABSTRACTWe validated an antigen capture enzyme-linked immunosorbent assay (ELISA) in Colombian persons with AIDS and proven histoplasmosis and evaluated the correlation between antigenuria and clinical improvement during follow-up. The sensitivity of theHistoplasma capsulatumELISA was 86%, and the overall specificity was 94%. The antigen test successfully monitored the response to therapy.


2012 ◽  
Vol 24 (5) ◽  
pp. 895-902 ◽  
Author(s):  
Jasmina Kircanski ◽  
Douglas Hodgins ◽  
Glenn Soltes ◽  
Yanlong Pei ◽  
Valeria R. Parreira ◽  
...  

An enzyme-linked immunosorbent assay (ELISA) was developed for detection and quantitation of beta2-toxin in neonatal piglet intestinal contents. Polystyrene plates were coated with polyclonal capture antibodies prepared against consensus recombinant beta2-toxin. The ELISA was developed using consensus recombinant beta2-toxin, atypical recombinant beta2-toxin, purified consensus native beta2-toxin, and field samples of neonatal porcine intestinal contents. Captured antigen was detected using a horseradish peroxidase–labeled monoclonal antibody against consensus recombinant beta2-toxin. The limit of detection of the ELISA for consensus beta2-toxin was between 2.0 and 3.5 ng/ml. The ELISA detected atypical recombinant beta2-toxin only weakly. Optical density was protein concentration dependent. The test confirmed differences between consensus and atypical recombinant beta2-toxin, but similar results obtained when testing pure consensus recombinant beta2-toxin and native beta2-toxin. Results obtained from intestinal content samples, particularly from the small intestine, were highly inconsistent and suggested variable protease activity. Addition of protease inhibitors partially prevented degradation of the toxin; however, sample processing at low temperature, at a lower pH (citrate buffer with 5% of bovine serum albumin, pH 6.1), and “cold incubation” of applied antigens abolished protease activity. The recombinant toxin was preserved in spiked intestinal samples by freezing at −70°C, suggesting that necropsy samples can be stored frozen for periodic testing. With appropriate sample preparation, antigen-capture ELISA can detect beta2-toxin in the intestinal content and feces of neonatal piglets.


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