胡佳莉,邓鹏,姜勇
HU Jiali,DENG Peng,JIANG Yong(Department of Pathophysiology of Southern Medical University and Key Laboratory of Proteomics of Guangdong Province

• 1:南方医科大学病理生理学教研室
• 2:广东省蛋白质组学重点实验室

摘要(Abstract)：

目的:在传统多重连接探针扩增(MLPA)技术基础上建立一种更为特异的检测单核苷酸多态性(SNP)位点的方法。方法:设计针对Kras基因216 G-C SNP位点的MLPA探针,使用不同方法优化MLPA实验,观察其对SNP位点检测时的影响。结果:直接用锁核酸(LNA)修饰MLPA探针的SNP位点或添加阻滞探针均可显著提高SNP检测时的特异性,3个LNA修饰的阻滞探针能最好地保证检测的灵敏度和特异性。结论:成功地建立了一种增强MLPA检测SNP位点的特异性方法。
Objective: To establish a more speicific method for detecting SNP site based on tranditional MLPA technique.Methods: MLPA probes for Kras gene 216 G-C SNP site were designed.Different methods were employed to optimize the MLPA experiment,and the influence of them on detecting SNP site was observed.Result: Either direct modification of SNP site on MLPA probe with LNA or adding blocking probes could significantly enhance the specificity in SNP detection.Three LNAs modified blocking probe could ensure the best sensitivity and specificity of the detection.Conclusions: A method for enhancing the specificity of MLPA in detecting SNP site is successfully established.

关键词(KeyWords)： 核酸探针;多态性,单核苷酸;敏感性与特异性
nucleic acid probes;polymorphism,single nucleotide;sensitivity and specificity

Abstract:

Keywords:

基金项目(Foundation): 国家973计划项目(2002CB513005);; 国家自然科学基金委员会-广东省人民政府自然科学联合基金重点项目(U0632004);; 教育部长江学者和创新团队发展计划项目(IRT0731)资助

作者(Author): 胡佳莉,邓鹏,姜勇
HU Jiali,DENG Peng,JIANG Yong(Department of Pathophysiology of Southern Medical University and Key Laboratory of Proteomics of Guangdong Province

DOI: 10.19367/j.cnki.1000-2707.2012.03.005

参考文献(References)：

• [1]汤桂丽,赵映兰.单核苷酸多态性检测方法的研究进展[J].中国药业,2010(7):1-2.
• [2]Schouten JP,McElgunn CJ,Paris G,et al.Relativequantification of 40 nucleic acid sequences by multiplexligation-dependent probe amplification[J].Nucleic AcidsRes,2002(12):e57.
• [3]Nygren AH,Ameziane A,Errami A,et al.Methylation-specific MLPA(MS-MLPA):simultaneous detection ofCpG methylation and copy number changes of up to 40 se-quences[J].Nucleic Acids Res,2005(14):e128.
• [4]Kozlowski P,Jasinska AJ,Kwiatkowski DJ.New applica-tions and development in the use of multiplex ligation-de-pendent probe amplification[J].Elecrophoresis,2008(23):4627-4636.
• [5]Morandi L,de Biase D,Visani M,et al.Allele specificlocked nucleic acid quantitative PCR(ASLNAqPCR):anaccurate and cost-effective assay to diagnose and quantifyKRAS and BRAF mutation[J].Plos One,2012(4):36084.
• [6]Didelot A,Le Corre D,Luscan A,et al.Competitive al-lele specific TaqMan PCR for KRAS,BRAF and EGFRmutation detection in clinical formalin fixed paraffin em-bedded samples[J].2012(3):275-280.

文章评论(Comment)：