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. 2020 Mar 6;13(1):135.
doi: 10.1186/s13104-020-04981-y.

Protein A/G-based enzyme-linked immunosorbent assay for detection of anti-Pythium insidiosum antibodies in human and animal subjects

Chalisa Jaturapaktrarak  1   2 , Penpan Payattikul  1 , Tassanee Lohnoo  1 , Yothin Kumsang  1 , Aree Laikul  3 , Watcharapol Pathomsakulwong  4 , Chompoonek Yurayart  5 , Walaiporn Tonpitak  6 , Theerapong Krajaejun  7
Affiliations

Protein A/G-based enzyme-linked immunosorbent assay for detection of anti-Pythium insidiosum antibodies in human and animal subjects

Chalisa Jaturapaktrarak et al. BMC Res Notes. .

Abstract

Objectives: Pythiosis is a deadly infectious disease caused by Pythium insidiosum. Reports of both human and animal pythiosis are on the rise worldwide. Prognosis of the pythiosis patients relies on early diagnosis and prompt treatment. There are needs for an immunodiagnostic test that can detect the disease in both humans and animals. This study aims at reporting an optimized protocol for the development of a protein A/G-based enzyme-linked immunosorbent assay (ELISA) for the detection of anti-P. insidiosum antibody in multiple host species.

Results: A total of 25 pythiosis and 50 control sera, obtained from humans, horses, dogs, cats, and cows, were recruited for the assay development. With a proper ELISA cutoff point, all pythiosis sera can ultimately be distinguished from the control sera. The successfully-developed protein A/G-based ELISA can detect the anti-P. insidiosum antibodies in serum samples of both humans and animals. It is a versatile, feasible-to-develop, and functional immunodiagnostic assay for pythiosis.

Keywords: ELISA; Immunodiagnosis; Protein A/G; Pythiosis; Pythium insidiosum.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Dot plot of the ELISA values (EV) of pythiosis (n = 25) and control (n = 50) serum samples examined by a the protein A/G-based and b the anti-human IgG antibody-based ELISAs. Different colors indicate the EVs of sera from different animal species. Dashed lines represent the optimal ELISA cutoff points (see Table 1)
Fig. 2
Fig. 2
ELISA values (Y-axis) of serial twofold dilutions (from 1:1600 to 1:102,400; X-axis) of a serum sample from a human (blue), a horse (orange), and a dog (gray) with pythiosis, and a healthy individual (served as a control; yellow). The working serum dilution for protein A/G-based ELISA testing is 1:1600 (see text for details)
See this image and copyright information in PMC

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