Van Hoeyveld Erna, Houtmeyers Frans, Massonet Caroline, Moens Leen, Van Ranst Marc, Blanckaert N, Bossuyt Xavier
Department of Laboratory Medicine, Immunology, Gasthuisberg University Hospital, Herestraat 49, B-3000 Leuven, Belgium.
J Immunol Methods. 2004 Apr;287(1-2):227-30. doi: 10.1016/j.jim.2004.01.025.
Structural point mutations in exon 1 at codons 52, 54 and 57 and a promotor polymorphism at -221 bp of the mannose-binding lectin (MBL) gene are associated with increased susceptibility to various infectious diseases. We developed a genotyping method based on the 5' nuclease (TaqMan) assay in combination with the use of minor-groove-binder (MGB) probes in screening for these mutations/polymorphisms. In contrast to conventional probes, MGB probes have a short length and can be used for detection of mutations that are in close proximity to each other, as is the case for the structural mutations in exon 1 of the MBL gene. Results obtained with the 5' nuclease assay using MGB probes were identical with results obtained with classical techniques such as restriction fragment length polymorphism, allele-specific PCR, and sequencing. In conclusion, the 5' nuclease assay using MGB probes is useful for large-scale screening of point mutations/polymorphisms, even when they are in close proximity.
甘露糖结合凝集素（MBL）基因第1外显子52、54和57密码子处的结构点突变以及-221 bp处的启动子多态性与多种传染病易感性增加相关。我们开发了一种基于5'核酸酶（TaqMan）分析并结合使用小沟结合剂（MGB）探针的基因分型方法，用于筛选这些突变/多态性。与传统探针不同，MGB探针长度较短，可用于检测彼此紧邻的突变，MBL基因第1外显子的结构突变就是这种情况。使用MGB探针的5'核酸酶分析获得的结果与使用经典技术（如限制性片段长度多态性、等位基因特异性PCR和测序）获得的结果一致。总之，使用MGB探针的5'核酸酶分析可用于大规模筛选点突变/多态性，即使它们彼此紧邻。
