二氧化锰纳米片介导的旋毛虫病检测方法的建立OA北大核心CSTPCD
Establishment of a detection method for trichinellosis mediated by MnO2 nanosheets
以半胱氨酸蛋白酶抑制剂样蛋白(Cystatin-like protein,CLP)为研究对象,以实验室前期制备的重组蛋白rCLP为检测抗原进行包被,抗CLP单克隆抗体(McAb)1H9作为竞争抗体,利用二氧化锰纳米片独特的光学特性,建立一种准确、快速、高效的旋毛虫病比色免疫测定方法.采用旋毛虫人工感染猪血清、其他寄生虫感染猪血清及常规病毒疫苗免疫猪血清对该方法的特异性、敏感性进行验证.结果显示,该方法检测刚地弓形虫、微小隐孢子虫、猪囊尾蚴、猪蛔虫、猪鞭虫感染猪的血清和常规病毒疫苗(猪伪狂犬病病毒、猪繁殖与呼吸综合征病毒、猪圆环病毒、猪瘟病毒和口蹄疫病毒疫苗)免疫猪血清均为阴性,无交叉反应.对高(10 000条/头)、中(1 000条/头)、低(100条/头)3组感染剂量感染后不同时间的人工感染猪血清进行检测,该方法在高剂量组中检测的血清抗体阳性转化时间与商品化试剂盒的检测结果一致,在中、低剂量组中血清抗体阳性转化时间晚于商品化试剂盒.同时,该方法在低剂量组中检测的旋毛虫感染阳性检出率高于人工消化法的检测结果,说明该方法敏感性较高.对所建立的方法进行加标回收验证,回收率为98.8%~102.7%,表明该方法稳定性高.以上结果表明,本研究建立的二氧化锰纳米片介导的旋毛虫病检测方法特异性好、敏感性高、操作简单、无须大型仪器即可出结果,可用于旋毛虫病的快速筛检.
This study focused on cystatin-like protein(CLP)as the research object,using the recom-binant protein rCLP prepared in the laboratory as the detection antigen for coating,and the anti-CLP monoclonal antibody(McAb)1H9 as the competitive antibody.Utilizing the unique optical properties of manganese dioxide nanosheets,an accurate,fast,and efficient colorimetric immunoassay for trichinello-sis was established.The specificity and sensitivity of this method were verified by using the serum of swine artificially infected with Trichinella spiralis,the serum of swine infected with other parasites,and the serum of swine immunized with conventional virus vaccines.The results showed that the serum of swine infected with Toxoplasma gondii,Cryptosporidium parvum,Cysticercus cellulosae,Ascaris suum,and Trichuris suis,and the sera of swine immunized with conventional virus vaccines(Vaccines for porcine pseudorabies virus,porcine reproductive and respiratory syndrome virus,porcine circovirus,swine fever virus,and foot-and-mouth disease virus)were all negative and no cross-reaction was observed.The serum samples of artificially infected pigs with three infection doses of different infection days were detected,including high(10 000 larvae/head),medium(1 000 larvae/head)and low(100 larvae/head)dose groups.The positive transformation time of serum antibody in the high-dose group was consistent with that of the commercial kit,and the positive transformation time of serum antibody in the medium and low dose groups was later than that of the commercial kit.At the same time,the positive detection rate of T.spiralis infection in the low dose group was higher than that of the artificial digestion method,indicating that the method was more sensitive.The established method was used for spiked recovery,the recovery rate was 98.8%—102.7%and the method was highly stable.The above results indicated that the proposed method for trichinellosis based on Mn02 nanosheets had high specificity,high sensitivity,simple operation,and can produce results without the large instruments.It can be used for on-site rapid screening of trichinellosis.
冯馨锐;邹敏鸿;徐凝;赵阳;李明娜;郑策;刘琰;赵臣
延边大学医学院,吉林延吉 133002||吉林医药学院公共卫生学院,吉林吉林 132013吉林医药学院公共卫生学院,吉林吉林 132013吉林大学人畜共患传染病重症诊治全国重点实验室人兽共患病研究教育部重点实验室人兽共患病研究所,吉林长春 130062中国人民解放军联勤保障部队第九六五医院急诊与重症医学科,吉林吉林 132011
畜牧业
旋毛虫半胱氨酸蛋白酶抑制剂样蛋白比色免疫测定二氧化锰纳米片
Trichinella spiral iscystat in-like proteincolorimetric immunoassayMn02 nanosheets
《中国兽医科学》 2024 (007)
921-929 / 9
国家自然科学基金项目(82302563);省级大学生创新创业项目(S202313706015)
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