Salazar M, Yunis J J, Delgado M B, Bing D, Yunis E J
Department of Immunogenetics, Dana Farber Cancer Institute, Boston, MA 02115.
Tissue Antigens. 1992 Sep;40(3):116-23. doi: 10.1111/j.1399-0039.1992.tb02102.x.
We have developed a new PCR-RFLP method for HLA-DQB1 typing. This method was easy to follow, requiring only one DQB1 generic amplification and 5 endonucleases to assign 14 out of 15 HLA-DQB1 alleles. In addition, we determined that by using one generic amplification and two enzymes (Sau96 I and Hae III) it was possible to type the generic specificities: DQw2, DQw4, DQw5, DQw6, and DQw7, DQw8-9, providing a practical alternative for serological HLA-DQ generic typing. We also performed a side-by-side correlation with a PCR-SSO typing method and found an almost 100% concordance between the methods. The limitations of these methods were: 1) the PCR-RFLP method did not allow the differentiation between the HLA-DQB1*0602 and *0603 alleles; 2) the PCR-SSO method gave crosshybridization signals in the detection of *0302 or *0303 alleles. Our results suggested that both methods, PCR-RFLP and PCR-SSO, are useful alternatives for HLA-DQB1 typing.
我们开发了一种用于HLA - DQB1分型的新型PCR - RFLP方法。该方法易于操作，仅需一次DQB1通用扩增和5种内切酶即可鉴定15种HLA - DQB1等位基因中的14种。此外，我们确定通过一次通用扩增和两种酶（Sau96 I和Hae III）可以对通用特异性进行分型：DQw2、DQw4、DQw5、DQw6以及DQw7、DQw8 - 9，为血清学HLA - DQ通用分型提供了一种实用的替代方法。我们还与PCR - SSO分型方法进行了并行相关性分析，发现两种方法之间的一致性几乎达到100%。这些方法的局限性在于：1）PCR - RFLP方法无法区分HLA - DQB10602和0603等位基因；2）PCR - SSO方法在检测0302或0303等位基因时出现交叉杂交信号。我们的结果表明，PCR - RFLP和PCR - SSO这两种方法都是HLA - DQB1分型的有用替代方法。
