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Immunodiagnosis of opisthorchiasis using parasite cathepsin F

Abstract

Opisthorchis viverrini, a food-borne trematode parasite endemic in the lower Mekong countries, is conventionally diagnosed by stool examination. However, parasitological stool-based diagnosis can be unreliable in light infections. The goal of this study was to develop the immunodiagnosis of opisthorchiasis using cathepsin F cysteine protease of O. viverrini in both indirect and sandwich ELISA assays. A recombinant O. viverrini cathepsin F (rOv-CF) of 40 kDa was expressed in E. coli strain BL21 (DE3), affinity purified, and deployed in ELISA assays. Human sera from 272 cases were investigated by indirect rOv-CF-based ELISA. Positive antibody response to rOv-CF was found in 137 out of 272 cases (50.37 %) using a cutoff OD (0.400) determined by ROC analysis. In comparison to parasitological stool examined for fluke eggs, the gold standard, the rOv-CF indirect ELISA showed a sensitivity and specificity of 62.1 and 84.05 %, respectively. Serum antibody levels correlated well with egg counts per gram feces (EPG) (P < 0.001). In addition, chicken IgY antibody raised against rOv-CF was tested in a sandwich ELISA for detection of coproantigen in the feces of experimentally infected hamsters. The sandwich ELISA using this chicken IgY in combination with rabbit antibody to O. viverrini somatic antigens showed sensitivity and specificity of 93.3 and 78.57 %, respectively. Together, these findings indicated the potential of rOv-CF for diagnosis of opisthorchiasis, including for uses with chicken IgY for detection of coproantigens of O. viverrini.

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Acknowledgments

We gratefully acknowledge support from the Higher Education Research Promotion and National Research University Project of Thailand, Office of the Higher Education Commission, through the Health Cluster (SHeP-GMS), Khon Kaen University, from the Thailand Research Fund (TRF) grant number RTA 5680006, from the National Institute of Allergy and Infectious Diseases (NIAID), NIH, grant number P50AI098639 and from the United States Army Medical Research and Materiel Command (USAMRMC), contract number W81XWH-12-C-0267. BS is a TRF Senior Scholar. The content is solely the responsibility of the authors and does not necessarily represent the official views of the USAMRMC, NIAID or the NIH or the funders.

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Correspondence to Banchob Sripa.

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Teimoori, S., Arimatsu, Y., Laha, T. et al. Immunodiagnosis of opisthorchiasis using parasite cathepsin F. Parasitol Res 114, 4571–4578 (2015). https://doi.org/10.1007/s00436-015-4703-9

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Keywords

  • Opisthorchis viverrini
  • Cathepsin F
  • Immunodiagnosis
  • Coproantigen detection
  • Opisthorchiasis
  • Food-borne trematodiasis