Saposin-like protein 2 has an immunodiagnostic potential for detecting Fasciolosis gigantica

Pornanan Kueakhai; Narin Changklungmoa; Kulathida Chaithirayanon; Manussabhorn Phatsara; Narin Preyavichyapugdee; Suda Riengrojpitak; Kant Sangpairoj; Phiraphol Chusongsang; Prasert Sobhon

doi:10.1016/j.exppara.2015.01.019

Saposin-like protein 2 has an immunodiagnostic potential for detecting Fasciolosis gigantica

Exp Parasitol. 2015 Apr-May:151-152:8-13. doi: 10.1016/j.exppara.2015.01.019. Epub 2015 Feb 3.

Authors

Pornanan Kueakhai¹, Narin Changklungmoa², Kulathida Chaithirayanon³, Manussabhorn Phatsara⁴, Narin Preyavichyapugdee⁵, Suda Riengrojpitak⁶, Kant Sangpairoj³, Phiraphol Chusongsang⁷, Prasert Sobhon⁸

Affiliations

¹ Department of Pathobiology, Faculty of Science, Mahidol University, Rama VI Rd, Ratchathewi, Bangkok 10400, Thailand; Faculty of Allied Health Sciences, Burapha University, Long-Hard Bangsaen Rd, Saen Sook Sub-district, Mueang District, Chonburi 20131, Thailand.
² Faculty of Allied Health Sciences, Burapha University, Long-Hard Bangsaen Rd, Saen Sook Sub-district, Mueang District, Chonburi 20131, Thailand.
³ Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd, Ratchathewi, Bangkok 10400, Thailand.
⁴ Department of Anatomy, Faculty of Medicine, Chiang Mai University, Chiang Mai 52000, Thailand.
⁵ Faculty of Animal Science, Silpakorn University, Phetchaburi Campus, Cha-am, Phetchaburi 76120, Thailand.
⁶ Department of Pathobiology, Faculty of Science, Mahidol University, Rama VI Rd, Ratchathewi, Bangkok 10400, Thailand.
⁷ Department of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, 10400, Thailand.
⁸ Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd, Ratchathewi, Bangkok 10400, Thailand. Electronic address: prasert.sob@mahidol.ac.th.

PMID: 25662434
DOI: 10.1016/j.exppara.2015.01.019

Abstract

Saposin-like protein 2 (SAP-2) plays an important role in the digestive process of Fasciola gigantica (Fg). It is one of the major proteins synthesized by the caecal epithelial cells and released into fluke's excretion-secretion. Therefore, FgSAP-2 is a plausible target for detecting fasciolosis. A polyclonal antibody (PoAb) against recombinant FgSAP-2 was produced by immunizing rabbits with the recombinant protein (rFgSAP-2), and used in sandwich ELISA assay to detect the circulating FgSAP-2 in sera of mice experimentally infected with F. gigantica metacercariae. The assay could detect rFgSAP-2 and the native FgSAP-2 in the excretory-secretory (ES) and whole body (WB) fractions of adult F. gigantica at the concentrations as low as 38 pg/ml, 24 ng/ml, and 102 ng/ml, respectively. As well, the sera from mice experimentally infected with F. gigantica were tested positive by this sandwich ELISA, which exhibited sensitivity, specificity, false positive rate, false negative rate and accuracy at 99.99, 98.67, 1.33, 0.01 and 99.32%, respectively. Therefore, this assay could be used for diagnosis of fasciolosis by F. gigantica.

Keywords: Fasciola gigantica; Immunodiagnosis; Sandwich ELISA; Saposin-like protein 2.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antibodies, Helminth / biosynthesis
Antibodies, Helminth / blood
Antibodies, Helminth / immunology
Antigens, Helminth / blood*
Enzyme-Linked Immunosorbent Assay / methods
Enzyme-Linked Immunosorbent Assay / standards*
False Negative Reactions
False Positive Reactions
Fasciola / immunology
Fasciola / isolation & purification*
Fasciola / metabolism
Fascioliasis / blood
Fascioliasis / diagnosis*
Immunoglobulin G / blood
Immunoglobulin G / isolation & purification
Male
Mice
Rabbits
Recombinant Proteins / immunology
Saposins* / immunology
Saposins* / metabolism
Schistosomiasis / blood
Schistosomiasis / diagnosis
Sensitivity and Specificity

Substances

Antibodies, Helminth
Antigens, Helminth
Immunoglobulin G
Recombinant Proteins
Saposins