HLA-DRB_1基因快速分型在眼库中的应用

The application of HLA DRB 1 genetic typing in an eye bank

目的建立一种快速准确的眼库ＨＬＡ基因分型方法。方法取眼库中新鲜或冷冻保存的供体眼球的球结膜、眼外肌、视神经、角巩膜缘、巩膜、虹膜、晶状体、玻璃体及视网膜等组织各约１０ｍｇ，快速提取ＤＮＡ，然后利用针对ＨＬＡＤＲＢ１区１３种抗原的组织特异性ＤＮＡ引物进行ＰＣＲ扩增，琼脂糖电泳基因分型。结果除晶状体、玻璃体、虹膜组织外，利用其它眼部组织都准确地进行了ＨＬＡＤＲＢ１区的基因分型，而且同一供体的不同组织分型结果完全一致，全部过程仅需５ｈ左右。结论ＨＬＡ基因分型方法快速、准确、特异性强，所需标本量少，眼库易得。

ObjectiveTo establish a quick,accurate HLA genetic typing method in an eye bank.Methods10 mg donor tissues of conjunctiva,rectus muscle,optic nerve,corneal rim,sclera,iris,lens and vitreous were obtained from the eye bank.The DNA samples were extracted from these tissues individually.PCR amplification was performed using 13 pairs of group specific HLA DRB 1 primers,and the reaction products were analysed on 2% agarose gels with ethidium bromide staining.ResultsWith the exception of iris,lens and vitreous,all samples were found to be suitable for HLA DRB 1 genetic typing.The typing results were consistent for tissues from the same donor.The whole procedure only needed about 5h.ConclusionHLA genetic typing is a rapid,accurate and highly specific method,and is suitable for clinical application.