猪蛔虫幼虫琼脂胶移行法的建立

Establishment of a Agar-gel migration assay for Ascaris suum larvae

本实验首次在国内建立起了线虫的琼脂胶移行法 (Agar- gel Migration Assay,AMA)。通过对影响幼虫活力的某些理化因素 ,如 :琼脂胶的温度、每孔加入的琼脂胶和幼虫混合液的体积、琼脂胶的浓度及幼虫在琼脂胶内移行的时间等研究发现 :加胶的温度和加胶的量对幼虫的移出率有较大的影响。幼虫的移出率随加胶温度的升高而降低 ,当温度达到 70℃时 ,则几乎无虫体从琼脂胶内移出 (移出率仅为 0 .5 % ) ;幼虫的移出率随加入琼脂胶量的增多而减小 ,当每孔加入 10 0 0μl时 ,幼虫的移出率为 16 .6 8% ,当加入 4 0 0μl时幼虫的移出率则能达到2 5 .17%。琼脂胶的浓度对幼虫移出的影响并不明显 ,采用 SAS软件 (for Windows V6 .12 )分析发现 1.2 %、1.0 %、0 .8%、0 .4 %的结果间无显著差异 (P>0 .0 5 ) ,但琼脂胶的浓度能影响胶液凝固的时间和凝固后的韧性。实验表明 :最适宜的加胶量为 4 0 0μl /孔 ,最适宜的加胶温度为 5 3℃ (Agar- gel,sigm a) ,在琼脂胶中最佳培养移行时间为 2 4 h,琼脂胶液的最适实验浓度为 1.5

This is the first time to set up the Agar gel migration assay (AMA) of nematode in China. We studied some physical and chemical factors which may affect the motility of the third stage larvae (L 3) of Ascaris suum , such as the temperature of agar gel, the volume of mixture (agar gel with L 3 of Ascaris suum) added into every well, the density of agar gel and the time in which L 3 migrated in agar gel. The result indicated that the percentage of larvae migrated (PLM) onto the surface of agar gel decreased as temperature and thickness of agar gel increased and was not influenced by the concentration of agar gel. The optimum condition for AMA of Ascaris suum L 3 are listed below. The temperature of agar gel is 53℃,the volume of mixture is 400 μl, the concentration of agar gel is 1.5% and larvae migration time is over 24 h .