摘要
目的:制备一种可无创、快速检测人体汗液中的白细胞介素-6(interleukin 6,IL-6)的免疫层析贴片,并对其性能进行评价。方法:采用柠檬酸三钠还原法制备胶体金,并对反应体系中的胶体金标记抗体的浓度以及pH值进行优化。采用免疫夹心法,以胶体金为显色媒介制备免疫层析贴片。制备的免疫层析贴片设计有亲水区域和疏水区域,其中,亲水区域包括亲水纸、样品垫、结合垫、硝酸纤维素(nitrocellulose,NC)膜和吸水纸;疏水区域包括疏水纸、疏水皮肤贴合胶、透明薄膜和聚氯乙烯(polyvinyl chloride,PVC)底板。采用制备的免疫层析贴片检测空白磷酸盐缓冲液(phosphate buffered saline,PBS)、人工汗液样品、真实汗液样品中的IL-6,以评价该免疫层析贴片的灵敏度和精准性。采用制备的免疫层析贴片对真实汗液样品中的IL-6、肿瘤坏死因子-α、新型冠状病毒和PBS进行检测,以评价该免疫层析贴片的特异性。结果:制备的免疫层析贴片可快速检出PBS、人工汗液样品、真实汗液样品中的IL-6,且对IL-6的最低检测限为20 pg/mL,灵敏度较高;该免疫层析贴片与肿瘤坏死因子-α、新型冠状病毒无交叉反应,具有良好的特异性。结论:制备的免疫层析贴片可实现无创、快速检测人体汗液中的IL-6,适用于临床检测。
Objective To prepare an immunochromatographic patch for non-invasive and rapid detection of interleukin 6(IL-6)in human sweat and evaluate its performance.Methods Colloidal gold was prepared by the trisodium citrate reduction method,and optimization was carried out for the concentration of colloidal gold-labeled antibody in the reaction system and the pH value.Then an immunochromatographic patch was manufactured by immunosandwich method using colloidal gold as the chromogenic medium,which was divided into a hydrophilic region and a hydrophobic region.There were hydrophilic paper,sample pads,binding pads,nitrocellulose(NC)membranes and absorbent paper involved in the hydrophilic region,and hydrophobic paper,hydrophobic skin-fit adhesive,transparent film and polyvinyl chloride(PVC)backing plate in the hydrophobic region.The prepared immunochromatographic patch had its sensitivity and precision evaluated by detecting the IL-6 in phosphate buffered saline(PBS),artificial sweat samples and real sweat samples,and its specificity by testing IL-6,TNF-α,SARS-CoV-2 and blank PBS in real sweat samples.Results The prepared immunochromatographic patch proved its high sensitivity by rapidly detecting IL-6 in PBS,artificial sweat and real sweat samples,with the lowest detection limit being 20 pg/mL for IL-6;the immunochromatographic patch had no cross-reactivity with TNF-αand SARS-CoV-2,and showed high specificity.Conclusion The prepared immunochromatography patch realizes non-invasive and rapid detection of IL-6 in human sweat,and can be used for clinical laboratory examinations.
作者
任梦
贺强
刘英帅
马萍
杨旭
REN Meng;HE Qiang;LIU Ying-shuai;MA Ping;YANG Xu(School of Pharmacy,Faculty of Medicine of Hubei University of Science and Technology,Xianning 437100,Hubei Province,China;Hubei Industrial Technology Research Institute of Intelligent Health,Xianning 437100,Hubei Province,China;School of Materials and Energy,Southwest University,Chongqing 400715,China;Xianning Public Inspection and Testing Center,Xianning 437100,Hubei Province,China)
出处
《医疗卫生装备》
CAS
2024年第12期32-38,共7页
Chinese Medical Equipment Journal
基金
咸宁市科技计划社发类研发重点专项(2023SF YF095,2021SFYF007)
湖北科技学院科研创新团队项目(2023T08)。
关键词
免疫层析技术
免疫夹心法
人体汗液
IL-6
免疫层析贴片
胶体金
immunochromatography
immunosandwich method
human sweat
IL-6
immunochromatographic patch
colloidal gold
