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人类白细胞抗原(HLA)及其检测方法简介。

The short story of HLA and its methods.

作者信息

Doxiadis Ilias I N, Claas Frans H J

机构信息

Department of Immunohaematology and Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands.

出版信息

Dev Ophthalmol. 2003;36:5-11. doi: 10.1159/000067651.

Abstract

BACKGROUND

During the past 40 years, typing for the human leukocyte antigens (HLA) was done using serological techniques. Several improvements were achieved as to the efficiency and reliability of these techniques. One of the major drawbacks of serology, however, is the need of viable cells. Especially blood cells drawn from deceased donors, as are the usual source for typing of corneal donors, may have a grossly impaired viability and a reduced expression of antigens on their cell surface. This makes serological typing difficult and liable to errors. In the mid-1980s, molecular typing was first introduced in many laboratories. This first period dealt with the so-called restriction fragment length polymorphism method, a tedious method not suited for prospective typing. Only with the introduction of the polymerase chain reaction was the suitability of molecular techniques with respect to perspective typing achieved.

METHODS

International workshops and the effort of many laboratories led to a standardization of the methods. External proficiency testing exercises on transplantation relevant procedures in the laboratories affiliated to transplantation centers and the introduction of an accreditation system in the USA and in Europe increased significantly the reliability of all relevant immunogenetical testing.

RESULTS

To date, patients and prospective organ and tissue donors are typed in addition to serology also with molecular methods. Using these techniques, the reliability and reproducibility of HLA typing have reached levels of more than 98%. Even 8-day-old peripheral blood samples can now be typed routinely with these methods, formerly impossible by serology.

CONCLUSION

The laboratories affiliated to the transplantation centers are ready for high reliability testing of all HLA markers required by the clinic according to the results of controlled clinical long-term follow-up studies.

摘要

背景

在过去40年中,人类白细胞抗原(HLA)分型采用血清学技术。这些技术在效率和可靠性方面取得了一些改进。然而,血清学的一个主要缺点是需要活细胞。特别是从已故供体采集的血细胞,通常是角膜供体分型的来源,其活力可能严重受损,细胞表面抗原表达降低。这使得血清学分型困难且容易出错。20世纪80年代中期,分子分型首次在许多实验室引入。第一阶段采用所谓的限制性片段长度多态性方法,这是一种繁琐的方法,不适合前瞻性分型。只有引入聚合酶链反应,分子技术在前瞻性分型方面的适用性才得以实现。

方法

国际研讨会以及许多实验室的努力促成了方法的标准化。移植中心附属实验室对与移植相关程序的外部能力验证活动,以及美国和欧洲引入的认证系统,显著提高了所有相关免疫遗传学检测的可靠性。

结果

迄今为止,除血清学外,患者以及潜在的器官和组织供体也采用分子方法进行分型。使用这些技术,HLA分型的可靠性和可重复性已达到98%以上。现在,即使是8天龄的外周血样本也可以用这些方法常规分型,而这在以前通过血清学是不可能的。

结论

根据长期对照临床随访研究结果,移植中心附属实验室已准备好对临床所需的所有HLA标记物进行高可靠性检测。

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