Davidson J A, Poulton K V
Transplantation Laboratory, Manchester Institute of Nephrology and Transplantation, Manchester Royal Infirmary, UK.
Eur J Immunogenet. 2001 Oct;28(5):545-51. doi: 10.1046/j.0960-7420.2001.00251.x.
We have developed a short-form SSP-based HLA-DP typing system for routine use adapted from a comprehensive HLA-DP typing method described by Gilchrist et at. (1998). Our short-form system detects 93 alleles, including the 18 most common HLA-DPB1 alleles and eight HLA-DPA1 alleles. The primer mixes described were tested using the PCR-SSP Manager (Bunce et al., 1998) database to confirm the specificity of selected primers, and to detect potentially ambiguous amplifications. This short-form HLA-DP typing system was validated using 50 fully typed samples obtained through the UCLA International DNA Exchange. All samples gave 100% concordance with the consensus type. Our laboratory routinely uses a PCR-SSP based system of 48 primer mixes for HLA-DRB and HLA-DQB typing. The advantage of the short-form HLA-DP typing system described here is that the additional 48 HLA-DP primer mixes required can be included on the second half of a 96-well format tray. This method now enables a full HLA class II typing at the level of allele group resolution in 2 1/2 h.
我们基于吉尔克里斯特等人(1998年)描述的全面HLA-DP分型方法,开发了一种用于常规用途的基于序列特异性引物(SSP)的短形式HLA-DP分型系统。我们的短形式系统可检测93个等位基因,包括18个最常见的HLA-DPB1等位基因和8个HLA-DPA1等位基因。使用PCR-SSP管理器(邦斯等人,1998年)数据库对所述引物混合物进行测试,以确认所选引物的特异性,并检测潜在的模糊扩增。使用通过加州大学洛杉矶分校国际DNA交换获得的50个完全分型的样本对这种短形式HLA-DP分型系统进行了验证。所有样本与一致类型的一致性均为100%。我们实验室常规使用基于PCR-SSP的系统,该系统有48种引物混合物用于HLA-DRB和HLA-DQB分型。此处描述的短形式HLA-DP分型系统的优点是,所需的额外48种HLA-DP引物混合物可以包含在96孔板托盘的后半部分。这种方法现在能够在2个半小时内实现等位基因组分辨率水平的完整HLA II类分型。