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Immunodiagnosis of paramphistomosis using monoclonal antibody-based sandwich ELISA for detection of Paramphistomum gracile circulating 16 kDa antigen

Published online by Cambridge University Press:  21 February 2017

PANAT ANURACPREEDA ,
KULLANID TEPSUPORNKUL  and
RUNGLAWAN CHAWENGKIRTTIKUL
PANAT ANURACPREEDA*
Affiliation:
Institute of Molecular Biosciences, Mahidol University, Salaya, Nakorn Pathom 73170, Thailand Division of Agricultural Science, Mahidol University, Kanchanaburi Campus, Saiyok, Kanchanaburi, Thailand
KULLANID TEPSUPORNKUL
Affiliation:
Division of Agricultural Science, Mahidol University, Kanchanaburi Campus, Saiyok, Kanchanaburi, Thailand
RUNGLAWAN CHAWENGKIRTTIKUL
Affiliation:
Department of Microbiology, Faculty of Science, Mahidol University, Rama VI Road, Bangkok, Thailand
*
*Corresponding author. Division of Agricultural Science, Mahidol University, Kanchanaburi Campus, Saiyok, Kanchanaburi 71150, Thailand. E-mail: Panat1@yahoo.com, panat.anu@mahidol.ac.th
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Summary

In this study, we have produced a monoclonal antibody (MoAb) against 16 kDa antigen of Paramphistomum gracile (16 kDaAgPg), and developed an accurate sandwich enzyme-linked immunosorbent assay (sandwich ELISA) for the detection of circulating 16 kDaAg in the serum and fecal samples from cattle naturally infected with P. gracile. MoAb 1D10 was immobilized on a microtitre plate, and the antigen in the samples was captured and detected with biotinylated rabbit anti-16 kDaAgPg antibody. The lower detection limit of sandwich ELISA was 3·5 pg mL−1, and no cross-reaction with other parasite antigens was evaluated. The reliability of the assay was examined using the serum and fecal samples from cattle naturally infected with P. gracile, Fasciola gigantica, Moniezia benedeni, Trichuris sp., Strongyloides sp., strongylids and non-infected animals. The sandwich ELISA showed the sensitivity, specificity and accuracy at 98·33, 100 and 99·55% (serum samples), and 96·67, 100 and 99·09% (fecal samples). Therefore, this detection method is a rapid and excellent potential assay for the accurate diagnosis of paramphistomosis.

Keywords

Paramphisomum gracileparamphistomosis16 kDa antigenmonoclonal antibodysandwich ELISAimmunodiagnosis
Type
Research Article
Information
Parasitology , Volume 144 , Issue 7 , June 2017 , pp. 899 - 903
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Copyright
Copyright © Cambridge University Press 2017 

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