Takeshima S-N, Matsumoto Y, Miyasaka T, Arainga-Ramirez M, Saito H, Onuma M, Aida Y
Viral Infectious Diseases Unit, RIKEN, Wako, Saitama, Japan.
Tissue Antigens. 2011 Sep;78(3):208-13. doi: 10.1111/j.1399-0039.2011.01708.x. Epub 2011 May 29.
Recently, two polymerase chain reaction sequence-based typing (PCR-SBT) methods were reported for the genotyping of the bovine leukocyte antigen (BoLA)-DRB3. One technique is a single PCR-SBT (sPCR-SBT) method that generates heterozygous sequences that are subsequently analyzed by the haplofinder program, while the other technique is a nested PCR-SBT (nPCR-SBT) method that allows the analysis of heterozygous sequences using the assign 400ATF software. In this study, these techniques were compared and then integrated to produce an improved genotyping method. The primer set used for sPCR-SBT was more accurate than those used for nPCR-SBT. Combining sPCR-SBT with the assign 400ATF software previously reported for nPCR-SBT enables rapid and accurate genotyping of a large number of DNA samples.
最近，报道了两种基于聚合酶链反应序列分型（PCR-SBT）的方法用于牛白细胞抗原（BoLA）-DRB3的基因分型。一种技术是单重PCR-SBT（sPCR-SBT）方法，该方法产生杂合序列，随后通过单倍型查找程序进行分析，而另一种技术是巢式PCR-SBT（nPCR-SBT）方法，该方法允许使用assign 400ATF软件分析杂合序列。在本研究中，对这些技术进行了比较，然后将它们整合以产生一种改进的基因分型方法。用于sPCR-SBT的引物组比用于nPCR-SBT的引物组更准确。将sPCR-SBT与先前报道的用于nPCR-SBT的assign 400ATF软件相结合，能够对大量DNA样本进行快速准确的基因分型。
