山西医科大学学报2026，Vol.57Issue(8)：938-945,8.DOI:10.13753/j.issn.1007-6611.2026.08.010
切向流超滤法辅助ELISA检测血清中A型肉毒毒素
Tangential flow filtration-assisted ELISA for detection of botulinum toxin type A in human serum
摘要
Abstract
Objective To establish tangential flow ultrafiltration(TFU)-assisted ELISA for enhancing the sensitivity of botulinum toxin type A(BoNT-A)detection in human serum.Methods TFU was employed to separate and purify BoNT-A from serum samples.The effects of various parameters on toxin extraction efficiency were systematically investigated to optimize the ultrafiltration condi-tions,including membrane material(including modified polyethersulfone,polyethersulfone,and mixed cellulose ester),hollow fiber inner diameter,molecular weight cut-off(MWCO),membrane surface area,feed flow rate,transmembrane pressure(TMP,including 24.13 kPa,134.45 kPa,and 189.61 kPa),and circulation rate.The optimized ultrafiltration pretreatment protocol was methodologi-cally validated and subsequently integrated with ELISA for the detection of low-concentration botulinum toxin type A in clinical serum samples.Results Modified polyethersulfone exhibited the best extraction efficiency for botulinum toxin,reaching 79.17%,which was significantly higher than that of the other two membrane materials(P<0.05).The highest recovery rate of botulinum toxin was achieved at a transmembrane pressure of 134.45 kPa.Influent flow rate,circulation rate,hollow fiber inner diameter,molecular weight cut-off,and membrane surface area had no significant effect on extraction efficiency(P>0.05).Methodological validation showed that the recovery rate of the established TFU-assisted ELISA for detecting BoNT-A in human serum ranged from 90.0%to 105.0%,with a coefficient of variation below 10%.When applied to clinical serum samples,TFU-assisted ELISA detected low-concentration BoNT-A that were undetectable by conventional ELISA alone.Conclusion This study successfully established a TFU-assisted ELISA method for the detection of BoNT-A in human serum.This approach significantly enhanced the detection sensitivity of conven-tional ELISA for low-concentration BoNT-A,offering a reliable and efficient technical platform for accurate quantification of trace BoNT-A in clinical serum samples.关键词
A型肉毒毒素/切向流超滤/中空纤维膜/ELISA/方法学验证/血清Key words
botulinum toxin type A/tangential flow filtration/hollow fiber membrane/ELISA/method validation/serum分类
医药卫生引用本文复制引用
王文秀,周伟,胡爽,邢荣荣,陈璇..切向流超滤法辅助ELISA检测血清中A型肉毒毒素[J].山西医科大学学报,2026,57(8):938-945,8.基金项目
国家自然科学基金项目(81973287) （81973287）
国家自然科学基金青年基金项目(22104076) （22104076）
