大片形吸虫病早期诊断夹心ELISA方法的建立及应用

doi:10.16431/j.cnki.1671-7236.2016.04.007

Abstract

Abstract: To establish a method for the diagnosis of Fascioliasis gigantica in early stage, five hybridoma cell lines were recovered and used for preparation of monoclonal antibodies.7D2 was used as a capture antibody and 7D1 as detection antibody.Coating antibody dilution was 1:6 400 (0.208 μg/mL), 5% nonfat milk was used as blocking solution and detection antibody dilution was 1:10 000 (0.200 μg/mL).The detection limit of the sandwich ELISA was 1:3 200 (0.156 μg/mL), with good specificity and stability, and there was no cross reaction with other kinds of parasite antigen.The results showed that the method provided the important conditions and theoretical basis for the diagnosis of Fascioliasis gigantica in early stage. This would save unnecessary economic losses to the livestock, and it had clinical application value.

Key words: early diagnosis; monoclonal antibody; Fascioliasis gigantica; sandwich ELISA

CLC Number:

R446.61

LI Xue, WEI Zhi-peng, ZHAO Ming-long, XU Xin-ting, LIU Yu, WU Wen-de, JIANG Wei, HUANG Tian, LONG Fei-xiang, CHEN Han-zhong. Development and Application of Sandwich ELISA for Diagnosis of Fascioliasis gigantica in Early Stage[J]. , 2016, 43(4): 899-905.

References

[1] 陈凤,杨慧,刘榆华,等.云南省宾川县片形吸虫病流行病学调查及慢性病例分析[J].中国寄生虫学与寄生虫病杂志,2014,32(6):422-424.
[2] 殷铭阳,周东辉,朱兴全,等.中国牦牛主要寄生虫病流行现状及防控策略[J].中国畜牧兽医,2014,41(5):227-230.
[3] 陈凤,方文,王尚位,等.人体大片形吸虫虫卵发育观察[J].热带病与寄生虫学,2014,12(2):106-107.
[4] 方文,陈凤,刘宏坤.阿苯达唑和三氯苯哒唑对人体大片形吸虫的驱虫效果比较[J].中国血吸虫病防治杂志,2014,26(1):106-108.
[5] 李雪,黄甜,陈汉忠,等.大片形吸虫幼虫的培育及其感染小鼠的研究[J].中国畜牧兽医,2014,41(12):244-250.
[6] 徐怀英,朱瑞良,赵宏坤.肝片吸虫病诊断与防治研究进展[J].中国兽医寄生虫病,2000,8(4):49-51.
[7] 张克琴,陈宝杰,张克志,等.宾川县片形吸虫感染疫情的调查报告[J].卫生软科学,2013,27(3):164-166.
[8] 罗伟,刘建兵,罗智敏,等.血吸虫病门诊患者IHA与ELISA法抗体检测结果[J].热带医学杂志,2012,12(12):1502-1503.
[9] 林丹丹,刘跃民,胡飞,等.日本血吸虫病常用诊断方法应用价值的评估Ⅰ IHA筛查法对血吸虫病疫区人群感染率的评价[J].中国血吸虫病防治杂志,2008,20(3):179-180.
[10] 汪奇志,汪峰峰,尹晓梅,等.ELISA与IHA在血吸虫病不同流行程度地区的现场筛查效果评价[J].热带病与寄生虫学2006,3:135-136.
[11] 法爱玲,张悦,李爱霞.ELISA法与胶体金法检测健康人群抗-HCV的实验研究[J].中国卫生检验杂志,2004,14(1):21.
[12] 黄维义,张为宇,Ben B A,等.不同方法及材料检测大片吸虫感染的比较试验[J].中国兽医寄生虫病,1998,6(3):3-6.
[13] 陈汉忠,蒋金书,李义平.应用电化学免疫传感器检测大片形吸虫抗原抗体反应的研究[J].畜牧兽医学报,2004,35(3):339-342.
[14] 曾晓飞,陈汉忠,韦英益,等.抗水牛伊氏锥虫抗原单克隆抗体的制备及其初步应用[A].中国畜牧兽医学会家畜寄生虫学分会第六次代表大会暨第十次学术研讨会论文集[C].2009.
[15] Ai L,Chen M X,Alasaad S,et al.Genetic characterization,species differentiation and detection of Fasciola spp.by molecular approaches[J].Parasit Vectors,2011,4:101.
[16] Bernal D,de la Rubia J E,Carrasco-Abad A M,et al.Identification of enolase as a plasminogen-binding protein in excretory-secretory products of Fasciola hepatica[J]. FEBS Letters,2004,563(1-3):203-206.
[17] 李晓栩,刘明如,陈汉忠,等.双抗体夹心ELISA检测大片形吸虫分泌排泄抗原方法的建立[A].中国畜牧兽医学会家畜寄生虫学分会第六次代表大会暨第十一次学术研讨会论文集[C].2011.
[18] 薛松果.单抗体夹心ELISA检测鸡新城疫病毒抗原的研究[D].合肥:安徽农业大学,2004.
[19] 鲍玉芳.大片形吸虫硫氧还蛋白过氧化物酶基因的原核表达及早期检测方法的研究[D].南宁:广西大学,2012.

Development and Application of Sandwich ELISA for Diagnosis of Fascioliasis gigantica in Early Stage

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