富集H5N1亚型禽流感病毒生物磁珠的制备

Development of bio-magnetic beads to enrichment H5N1 influenza virus

制备了一种可以从大体积、低浓度病毒液体样品中富集H5N1亚型禽流感病毒的新型生物磁珠。比较了氨基磁珠与羧基磁珠作为生物磁珠载体的效果。结果表明,以pH 4.5MES(2-(N-吗啉)乙磺酸)为活化缓冲液活化羧基磁珠每毫克可以结合9.8μg的胎球蛋白,pH 4.5PBS(磷酸盐缓冲液)为活化缓冲液每毫克可以结合9.25μg的胎球蛋白;以5%戊二醛为活化剂活化氨基磁珠1h较以2.5%、7.5%的戊二醛为活化剂每毫克结合胎球蛋白量多,为8.12μg。制备完成的羧基型生物磁珠与氨基型生物磁珠均可富集H5N1亚型禽流感病毒,而羧基型生物磁珠可以富集检测出15倍病毒稀释液,敏感性较氨基磁珠富集检测出7倍病毒稀释液高。

This study was development a new bio-magnetic beads to enrichment H5N1 strain of avian influenza virus,which in a large volume or low-concentration liquid samples.Compared the amino group with the carboxyl magnetic beads.The results showed that carboxyl magnetic beads activated by pH 4.5MES（2-（N-morpholino）ethanesulfonic acid）can be combined with 9.8μg of fetuin（per mg）,pH 4.5PBS（phosphate buffered saline）as the activator buffer solution can be combined with 9.25μg of fetuin（per mg）;5%glutaraldehyde as amino magnetic beads activator which effected for 1hwas better than 2.5% and 7.5% glutaraldehyde,which can be combined with 8.12μg of fetuin（per mg）.Both the two type of bio-magnetic beads can enrich H5N1 strain of avian influenza virus.The carboxyl magnetic beads can enriched and detected 14-fold virus dilutions and the amino is 5-fold.