摘要
为制备抗阪崎肠杆菌(ES)脂多糖(LPS)抗原单克隆抗体(MAb)及建立双抗夹心ELISA检测方法,本研究以热灭活的ES(ATCC51329)菌体抗原作为免疫原,ES菌体结合LPS作为筛选抗原,采用杂交瘤细胞技术筛选获得9株稳定分泌抗ES LPS特异性MAb的杂交瘤细胞株。采用改良的过碘酸钠法制备辣根过氧化物酶(HRP)酶标抗体,并建立检测ES的双抗体夹心ELISA方法,该方法对ES ATCC51329具有良好的特异性,最低检测限可达103cfu/mL,为食品中ES的检测提供特异性MAb及ELISA检测方法。
To primarily establish the sandwich ELISA for Enterobacter sakazakii detection, Nine monoclonal antibodies (MAbs) against lipopolysaccharide (LPS) of E.sakazakii were prepare by fusion of SP2/0 myeloma cells with spleen cells fi'om BALB/c mice immunized with the E.sakazakii and scanned by the indirect ELISA coating with purified LPS from E.sakazakii. Meanwhile, the double sandwich ELISA was primarily developed for detecting E.sakazakii using a MAb conjugated with horseradish peroxidase (HRP) by using an improved sodium periodate oxidation method as the detection antibody, and other MAbs as capture antibody. The assay established in the present study was specific for E.sakazakii with the detection limit of 103 cfu/mL in pure culture of E.sakazakii ATCC51329. The results show that the specific MAbs and the sandwich-ELISA could be useful in the detection of E.sakazakii.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2013年第2期155-159,共5页
Chinese Journal of Preventive Veterinary Medicine