Tong J Y, Hammad A, Rudert W A, Trucco M, Hsia S
Department of Laboratory Medicine, Allegheny General Hospital, Pittsburgh, PA 15212.
Transplantation. 1994 Mar 15;57(5):741-5. doi: 10.1097/00007890-199403150-00018.
DNA heteroduplex (HD) electrophoretic patterns of DQB1 alleles from 124 individuals (38 members from 7 families and 43 kidney donor-recipient pairs) were analyzed in reference to each individual's DQB1 diallelic types determined by the polymerase chain reaction-RFLP method. The assignment of DQB1 homozygosity and heterozygosity, based solely on HD patterns, was accurate and correlated well with the typing results. DQB1 homozygotes invariably gave HD patterns of a single band while heterozygotes gave HD patterns of multiple bands. Distinct HD patterns of 2 heterozygotes predict the presence of at least 1 different DQB1 type between the pair. However, pairs with identical HD patterns may have different subtypes, because HDs with 1 or 2 nucleotide differences may sometimes give an identical HD pattern. Because of its simplicity and reproducibility, this HD analysis protocol serves as an excellent alternative to screen for DQB1 homozygotes and mismatched tissue donor-recipient pairs. This protocol is also useful for confirming the correctness of DQB1 allelic type assignments in a clinical setting.
参照通过聚合酶链反应-限制性片段长度多态性方法确定的每个个体的DQB1双等位基因类型,分析了124名个体(来自7个家庭的38名成员和43对肾脏供受者)的DQB1等位基因的DNA异源双链(HD)电泳图谱。仅基于HD图谱对DQB1纯合性和杂合性的判定是准确的,并且与分型结果高度相关。DQB1纯合子总是呈现单一条带的HD图谱,而杂合子则呈现多条带的HD图谱。两个杂合子不同的HD图谱预示这一对个体之间至少存在1种不同的DQB1类型。然而,具有相同HD图谱的配对可能具有不同的亚型,因为具有1个或2个核苷酸差异的异源双链有时可能给出相同的HD图谱。由于其简单性和可重复性,这种HD分析方法是筛选DQB1纯合子和不匹配的组织供受者配对的绝佳替代方法。该方法对于在临床环境中确认DQB1等位基因类型判定的正确性也很有用。