Evaluation of the Stability of Coated Plates with Antigen at Different Temperatures and Times by ELISA Test to Diagnose Fasciolosis
,
Abstract
Background: Considering that ELISA method presently is the test of choice for diagnosis of fasciolosis, the present study was undertaken to evaluate the maximum validity of coated plates at different temperatures and different times during one year of evaluation.
Methods: Serum samples of patients infected with fasciolosis (n=10), hydatidosis (n=5), toxocariasis (n=5), and negative control sera (n=5) were examined. Two series of plates were considered. The first series were coated with Fasciola homogenate Ag 12 ug/ml, and after some steps were blocked with gelatin and preserved at different temperatures as -80 °C , -20 °C, -4 °C and +4°C. The 2nd series were treated under the same criteria but were not blocked with gelatin. Each series were examined by ELISA test from 1st month to 12th month. Sera with 1:125 dilution, and peroxidase-conjugated goat anti-human IgG diluted 1:10000 were considered optimum.
Results: To ease reporting the results and due to many similarities only results related to 1st, 6th and 12th months were analyzed and sensitivity, specificity plus cut-off were determined for each series separately.
Conclusion: Preserving the coated plates, while unblocked at -80°C for 6-8 months is pertinent and functional and in that case, we can be sure the best out put would be applicable.
Mas-Coma S, Bargues MD, Valero MA. Fascioliasis and other plant-borne trematode zoonoses. Inter J Parasitolo. 2005;35:1255-1278.
Muller R. Worms and human diseases. 2nd ed. Wallingord: CABI International, Oxon, UK, 2002.
Hillyer GV. Immunodiagnosis of human and animal fasciolosis. In: Dalton JP, ed. Fasciolosis. Oxon: CABI Publishing, Wallingford, UK, 1998:435-48.
Rokni MB, Massoud J, O'Neill SM, Parkinson M, Dalton JP. Diagnosis of human fasciolosis in the Gilan province of Northern Iran: application of cathepsin L-ELISA. Diagn Microbiol Infect Dis. 2002;44:175-9.
Hillyer GV, Sanchez Z, DeLeon D. Immunodiagnosis of bovine fascioliasis by enzyme-linked immunosorbent assay and immunoprecipitation methods. J Parasitol. 1985;71:449-54.
O'Neill SM, Parkinson M, Dowd AJ, Strauss W, Angles R, Dalton JP. Short report: Immunodiagnosis of human fascioliasis using recombinant Fasciola hepatica cathepsin L1 cysteine proteinase. Am J Trop Med Hyg. 1999;60:749-751.
Rokni MB, Massoud J, Hanilo A. Comparison of adult somatic and cysteine proteinase antigens of Fasciola gigantica in enzyme linked immunosorbent assay for serodiagnosis of human fasciolosis. Acta Trop. 2003;88:69-75.
O’Neill SM, Parkinson M, Strauss W, Angles R, Dalton JP. Immu nodiagnosis of Fasciola hepatica infection (fasciolosis) in a human population in the Bolivian Altiplano using purified cathepsin L cysteine proteinase. Am J Trop Med Hyg. 1998;58:417-23.
Hillyer GV, Soler de Galanes M. Initial feasibility studies of the fast-ELISA for the immunodiagnosis of fascioliasis. J Parasitol.1991;77:362-5.
Galen RS. Predictive values & efficiency of laboratory testing. Pediater Clin North Am 1980;27:861-689.
Rokni MB, Lesan S, Massoud J, Kia EB, Molawi G. Comparative evaluation of Fast enzyme linked immunosorbent assay (Fast-ELISA) and standard-ELISA for the diagnosis of human hydatidosis. Iranian J Publ Health. 2006;35:29-32.
Sadjjadi SM, Abidi H, Sarkari B, Izadpanah A, Kazemian S. Evaluation of Enzyme Linked Immunosorbent Assay, Utilizing Native Antigen B for Serodiagnosis of Human Hydatidosis. Iran J Immunol. 2007;4 167-172.
Koosha S, Fesharaki M, Rokni MB. Comparison of enzyme-linked immunosorbent assay and indirect immunofluorescence assay in the diagnosis of human strongyloidiasis. Indian
| Files | ||
| Issue | Vol 5 No 1 (2010) | |
| Section | Articles | |
| Keywords | ||
| Fasciolosis ELISA Plate Validity Sensitivity Specificity | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
