基因Ⅱ型非洲猪瘟病毒(HLJ/18株)阳性血清定性标准样品研制

Preparation of qualitative reference material for positive serum specific for genotype Ⅱ African swine fever virus strain HLJ/18

摘要:

目的 研制基因Ⅱ型非洲猪瘟病毒(African swine fever virus, ASFV)阳性血清定性标准样品，为该病的血清学检测、质控及实验室检测能力验证等工作提供实物标准。方法 以灭活的基因Ⅱ型ASFV (HLJ/18株)免疫无特定病原体(specific pathogen-free, SPF)猪所获得的阳性血清为原料制备标准样品，采用间接酶联免疫吸附试验(indirect enzyme-linked immunosorbent assay, iELISA)等方法对该标准样品的纯净性、特异性、均匀性和稳定性等进行检验与评估，并委托9家实验室进行协作定值，3家实验室进行试用。结果 成功研制出500瓶基因Ⅱ型ASFV (HLJ/18株)阳性血清定性标准样品，该标准样品纯净、特异、均匀，在-20 ℃条件下可稳定保存至少18个月，在4、25和37 ℃均可至少稳定保存7 d。9家实验室协作定值结果一致，均判定为ASFV抗体阳性。结论 本研究成功研制了基因Ⅱ型ASFV (HLJ/18株)阳性血清定性标准样品，为该病检测相关工作提供了关键材料。

Abstract:

Objective To prepare reference material (RM) for positive serum specific for genotype Ⅱ African swine fever virus (ASFV) for serological detection, quality control, and proficiency testing (PT).Methods The anti-serum collected from specific pathogen-free (SPF) swine immunized with inactivated genotype Ⅱ ASFV was used as raw material for the preparation of RM. Indirect enzyme-linked immunosorbent assay (iELISA) was employed to evaluate the purity, specificity, homogeneity, and stability of RM. In addition, RM was characterized by nine laboratories and applied in clinical trials by three laboratories.Results A total of five hundred bottles of RM for positive serum specific for genotype Ⅱ ASFV strain HLJ/18 were successfully prepared. The results indicated that the RM we prepared was pure, homogenous, and free of exogenous virus contamination, showing good specificity, homogeneity, and stability. The RM was stable for at least 18 months when it was stored at -20 ℃ and for at least 7 days at 4 ℃, 25 ℃, and 37 ℃. The characterization by the nine laboratories showed that the RM was positive for antibodies against genotype Ⅱ ASFV.Conclusion The positive serum specific for genotype Ⅱ ASFV strain HLJ/18 has successfully been prepared, providing critical material for ASF detection and diagnosis.