应用MALDI-TOF质谱技术检测多发性骨髓瘤Jak2基因单核苷酸多态性

Detection of Single Nucleotide Polymorphisms of Jak2 Gene in Multiple Myeloma by Matrix-assisted Laser Desorption/ionization Time-of-Flight Mass Spectrometry

本研究探讨基质辅助激光解吸电离飞行时间质谱技术(MALDI-TOFMS)检测多发性骨髓瘤(MM)jak2基因的单核苷酸多态性(SNP)的实用性。用PCR扩增出含jak2基因2个SNP(C428T和C643T)位点的DNA片段作为模板,产物纯化后利用MALDI-TOFMS检测5例MM和5例健康对照者样本的jak2基因多态性。结果表明:C428T和C643T位点基因型在多发性骨髓瘤病例与健康对照组的分布无差异,均为T/T纯合子。结论:本实验建立的基于MALDI-TOFMS与引物延伸技术检测jak2基因多态性的方法是一个快速、准确和可靠的检测方法。

This study was purposed to investigate the practicality of matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF MS ） for detection of single nucleotide polymorphisms（SNP） on jak2 gene in multiple myeloma （MM） cells. The DNA fragment containing 2 SNPs of jak2 （C428T and C643T） was amplified using PCR and was purified. The purified product was used as the template for primer extension （PEX）. The small products of allele-specific reaction were purified, the SNPs on jak2 gene of 5 patients with MM and 5 healthy persons were detected by MALDI-TOF MS. The results showed that the distribution of genotype C428T and C643T was not different between MM patients and healthy persons, both of which are hamozygous T/T. In conclusion, the method based on MALDI-TOF MS and PEX technique for detecting SNP in jak2 gene is rapid, accurate and reliable method, and can be used in clinical practise.