Jiang-Baucom P, Girard J E, Butler J, Belgrader P
Department of Chemistry, American University, Washington, D.C. 20016, USA.
Anal Chem. 1997 Dec 1;69(23):4894-8. doi: 10.1021/ac970639u.
A novel analytical method using PNA probes detected by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOFMS) was applied to type sequence polymorphisms within the human leukocyte antigen (HLA), DQA locus. Streptavidin-coated magnetic beads were used to immobilize biotinylated DNA. PNA probes representing possible alleles were then prepared for the immobilized DNA hybridization. The nonspecific PNA probes were removed with stringent washes. The PNA/DNA/beads conjugate was analyzed by MALDI-TOFMS. The genotype of the DNA was determined by the detected molecular masses of the released PNA probes. Reproducible and accurate genotyping was achieved by this analytical method.
一种使用基质辅助激光解吸电离飞行时间质谱（MALDI-TOFMS）检测的肽核酸（PNA）探针的新型分析方法被应用于人类白细胞抗原（HLA）DQA基因座内的序列多态性分型。用链霉亲和素包被的磁珠固定生物素化的DNA。然后制备代表可能等位基因的PNA探针用于固定化DNA杂交。通过严格洗涤去除非特异性PNA探针。通过MALDI-TOFMS分析PNA/DNA/磁珠复合物。通过检测释放的PNA探针的分子量来确定DNA的基因型。通过这种分析方法实现了可重复且准确的基因分型。
