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

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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.

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Pythium insidiosum: an overview.

Wim Gaastra, Len Lipman, Arthur de Cock … (2010)

Pythium insidiosum is an oomycete pathogenic in mammals. The infection occurs mainly in tropical and subtropical areas, particularly in horses, dogs and humans. Infection is acquired through small wounds via contact with water that contains motile zoospores or other propagules (zoospores or hyphae). The disease, though described as emerging has in fact already been described since 1884. Depending on the site of entry, infection can lead to different forms of pythiosis i.e. a cutaneous, vascular, ocular, gastrointestinal and a systemic form, which is rarely seen. The infection is not contagious; no animal-animal or animal-human transmission has been reported so far. Therapy includes radical surgery, antifungal drugs, immunotherapy or a combination of these therapies. The prevention to contract the disease in endemic areas is difficult. Avoiding stagnant waters could be of help, although the presence of P. insidiosum on grass and soil in enzootic areas renders this practice useless.

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Clinical and epidemiological analyses of human pythiosis in Thailand.

Theerapong Krajaejun, Boonmee Sathapatayavongs, Roongnapa Pracharktam … (2006)

Pythiosis is an emerging and life-threatening infectious disease in humans and animals that is caused by the pathogenic oomycete Pythium insidiosum. Human pythiosis is found mostly in Thailand, although disease in animals has been increasingly reported worldwide. Clinical information on human pythiosis is limited, and health care professionals are unfamiliar with the disease, leading to underdiagnosis, delayed treatment, and poor prognosis. To retrospectively study the clinical and epidemiological features of human pythiosis, we analyzed clinical data from patients with pythiosis diagnosed during the period of January 1985 through June 2003 at 9 tertiary care hospitals throughout Thailand. A total of 102 cases of human pythiosis were documented nationwide. A substantial proportion (40%) of cases occurred in the last 4 years of the 18-year study interval. Clinical presentations fell into 4 groups: cutaneous/subcutaneous cases (5% of cases), vascular cases (59%), ocular cases (33%), and disseminated cases (3%). Almost all patients with cutaneous/subcutaneous, vascular, and disseminated pythiosis (85%) had underlying thalassemia-hemoglobinopathy syndrome. Most ocular cases (84%) were associated with no underlying disease. A majority of the patients were male (71%), were aged 20-60 years (86%), and reported an agricultural occupation (75%). Regarding treatment outcomes, all patients with disseminated infection died; 78% of patients with vascular disease required limb amputation, and 40% of these patients died; and 79% of patients with ocular pythiosis required enucleation/evisceration. Here, we report, to our knowledge, the largest case study of human pythiosis. The disease has high rates of morbidity and mortality. Early diagnosis and effective treatment are urgently needed to improve clinical outcomes. Because P. insidiosum is distributed worldwide and can infect healthy individuals, an awareness of human pythiosis should be promoted in Thailand and in other countries.

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Draft Genome Sequence of the Pathogenic Oomycete Pythium insidiosum Strain Pi-S, Isolated from a Patient with Pythiosis

Thidarat Rujirawat, Preecha Patumcharoenpol, Tassanee Lohnoo … (2015)

Pythium insidiosum is an oomycete that causes a life-threatening infectious disease called pythiosis in humans and animals living in tropical and subtropical countries. Here, we report the first draft genome sequence of P. insidiosum. The genome of P. insidiosum is 53.2 Mb and contains 14,962 open reading frames.

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Author and article information

Contributors

Theerapong Krajaejun:

ORCID: http://orcid.org/0000-0003-0545-3765

mr_en@hotmail.com

Journal

Journal ID (nlm-ta): BMC Res Notes

Journal ID (iso-abbrev): BMC Res Notes

Title: BMC Research Notes

Publisher: BioMed Central (London )

ISSN (Electronic): 1756-0500

Publication date (Electronic): 6 March 2020

Publication date PMC-release: 6 March 2020

Publication date Collection: 2020

Volume: 13

Electronic Location Identifier: 135

Affiliations

[1 ]GRID grid.10223.32, ISNI 0000 0004 1937 0490, Research Center, Faculty of Medicine, Ramathibodi Hospital, , Mahidol University, ; Bangkok, Thailand

[2 ]GRID grid.10223.32, ISNI 0000 0004 1937 0490, Molecular Medicine Program, Multidisciplinary Unit, Faculty of Science, , Mahidol University, ; Bangkok, Thailand

[3 ]GRID grid.9723.f, ISNI 0000 0001 0944 049X, Department of Large Animal and Wildlife Clinical Sciences, Faculty of Veterinary Medicine, , Kasetsart University, ; Nakhon Pathom, Thailand

[4 ]GRID grid.9723.f, ISNI 0000 0001 0944 049X, Equine Clinic, , Kasetsart University Veterinary Teaching Hospital, ; Nakhon Pathom, Thailand

[5 ]GRID grid.9723.f, ISNI 0000 0001 0944 049X, Department of Microbiology and Immunology, Faculty of Veterinary Medicine, , Kasetsart University, ; Bangkok, Thailand

[6 ]GRID grid.443723.5, Department of Microbiology, Faculty of Veterinary Medicine, , Mahanakorn University of Technology, ; Bangkok, Thailand

[7 ]Department of Pathology, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand

Author information

Theerapong Krajaejun http://orcid.org/0000-0003-0545-3765

Article

Publisher ID: 4981

DOI: 10.1186/s13104-020-04981-y

PMC ID: 7059324

PubMed ID: 32143691

SO-VID: caa380c4-c59a-4b5b-8a5d-1f015d0a0b24

License:

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

History

Date received : 24 January 2020

Date accepted : 26 February 2020

Funding

Funded by: FundRef http://dx.doi.org/10.13039/501100004396, Thailand Research Fund;

Award ID: RSA6280092

Award Recipient : Theerapong Krajaejun

Custom metadata

ScienceOpen disciplines: Medicine

Keywords: pythium insidiosum,pythiosis,immunodiagnosis,elisa,protein a/g

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ScienceOpen disciplines: Medicine

Keywords: pythium insidiosum, pythiosis, immunodiagnosis, elisa, protein a/g

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

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cGMP: Generators, Effectors and Therapeutic Implications Conference

Most cited references 26

Pythium insidiosum: an overview.

Clinical and epidemiological analyses of human pythiosis in Thailand.

Draft Genome Sequence of the Pathogenic Oomycete Pythium insidiosum Strain Pi-S, Isolated from a Patient with Pythiosis

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