摘要： 利用纯化的猪囊尾蚴重组蛋白M13h作为抗原包被酶标板,通过棋盘滴定方法筛选最佳反应条件,初步建立了猪囊尾蚴病间接ELISA检测方法。确定最佳包被量为1μg/mL,待检血清最佳稀释倍数为1∶100,酶标二抗的稀释倍数为1∶1 000,二者的作用时间均为45 min;与猪弓形虫病、猪旋毛虫病、细颈囊尾蚴病、猪蛔虫病、包虫病阳性血清均不发生反应,且具有较好的重复性。为囊虫病的流行病学调查提供了一种简便的血清学诊断方法。
关键词:
猪囊尾蚴,
M13h蛋白,
间接ELISA
Abstract: An indirect-ELISA was developed based on a recombinant protein M13h of Cysticercus cellulosae.The condition was selected successfully by phalanx titration.The ELISA used an optimal amount of the M13h protein at 1 μg/mL and a serum dilution of 1∶ 100,followed by adding rabbit anti-pig IgG HRP conjugate at 1∶ 1 000 and all incubated for 45 min.The recombinant antigen was highly specific and had no cross reactions with positive serum of toxoplasmosis,trichinelliasis,echinococciasis,ascariasis,cysticercus tenuicollis.Then it had a good repeatability.This indirect ELISA assay would provide a simple and rapid serodiagnosis means for epidemiological survey of cysticercosis.
Key words:
Cysticercus cellulosae,
M13h protein,
Indirect-ELISA
中图分类号:
王秋霞, 马景周, 宁长申, 张龙现, 菅复春. 基于猪囊尾蚴M13h蛋白的ELISA方法的优化[J]. 华北农学报, 2011, 26(4): 72-76. doi: 10.7668/hbnxb.2011.04.012.
WANG Qiu-xia, MA Jing-zhou, NING Chang-shen, ZHANG Long-xian, JIAN Fu-chun. Expression of Cysticercus cellulosae M13h Protein and Optimization of ELISA[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(4): 72-76. doi: 10.7668/hbnxb.2011.04.012.