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作 者:谢立信[1] 李绍伟[1] 董晓光[1] 袁凤波[1]
机构地区:[1]山东省医学科学院眼科研究所
出 处:《眼科研究》1998年第3期215-216,共2页Chinese Ophthalmic Research
摘 要: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HLADRB1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HLADRB1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。ObjectiveTo establish a quick,accurate HLA genetic typing method in an eye bank.Methods10 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.ResultsWith 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.ConclusionHLA genetic typing is a rapid,accurate and highly specific method,and is suitable for clinical application.
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