Establishment of an Experimental Procedure for Preparing Trial Serum Samples for the Specific Serodiagnosis of Toxocara canis for External Quality Assessment Schemes.

Autor: Vu, Quang Huy, Tran, Diep Tuan, Tran, Phu Manh Sieu, Le, Van Chuong, Huynh, Thi Diem Phuc, Bui, Quang Sang
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Zdroj: Journal of Parasitology Research; 2/10/2020, p1-7, 7p
Abstrakt: 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. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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