Establishment of an Experimental Procedure for Preparing Trial Serum Samples for the Specific Serodiagnosis of Toxocara canis for External Quality Assessment Schemes
| Autor: | Van Chuong Le, Quang Huy Vu, Thi Diem Phuc Huynh, Quang Sang Bui, Phu Manh Sieu Tran, Diep Tuan Tran |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Veterinary medicine biology Hbs antigen Article Subject 030231 tropical medicine Positive reaction Infectious and parasitic diseases RC109-216 030108 mycology & parasitology biology.organism_classification Serum samples Freezing methods 03 medical and health sciences 0302 clinical medicine Infectious Diseases Canis External quality assessment biology.protein Parasitology Antibody Toxocara canis |
| Zdroj: | Journal of Parasitology Research, Vol 2020 (2020) |
| ISSN: | 2090-0023 |
| DOI: | 10.1155/2020/6842975 |
| Popis: | Background. External quality assessment (EQA) provides evidence of reliable, accurate, and precise results for customers using the diagnostic test for Toxocara canis. Objective. To establish a procedure for producing standard Toxocara canis serum samples for serodiagnostic testing in EQA. Methods. The collected serum samples to contain anti-Toxocara canis antibodies were screened by ELISA and confirmed by Western blotting. These samples were found to be negative for other helminth antibodies, anti-HIV-1 and -2 antibodies, anti-HCV antibodies, and antibodies to HBs antigen. The sera were divided, processed by both freeze-drying and freezing methods, and then stored. The stability and homogeneity of the samples were evaluated after 7 days, 1 month, 3 months, and 6 months. An F-test and a T-test were applied to evaluate their homogeneity and stability. Results. Among eleven samples positive by ELISA, ten of them were confirmed via Western blotting by positive reaction with 5 specific Toxocara canis bands. Two lots of trial standard sera containing specific anti-Toxocara canis antibodies were successfully produced. Lot DK had a concentration of 31.01±1.1 NovaTec Units (NTU), and Lot DL had a concentration of 27.18±0.9 NTU. After storage at -80°C, the samples prepared by the freeze-drying method were stable for at least 3 months, and the samples prepared by the freezing method were stable for 6 months (p>0.05). Samples produced by both methods were stable for 7 days at 30°C (p>0.05). Conclusion. Specific serodiagnosis samples of anti-Toxocara canis antibodies for EQA could be produced that possessed homogeneity and stability lasting for 3 months and 6 months by the freeze-drying and freezing methods, respectively. At 30°C, the samples produced by both methods were stable for 7 days, suitable for delivery to remote laboratories. |
| Databáze: | OpenAIRE |
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