Rapid and Sensitive Detection of KRAS Mutation After fast-COLD-PCR Enrichment and High-resolution Melting Analysis
文献类型：期刊论文
| 作者 | Song, Chen1,2,3; Milbury, Coren A.1,2; Li, Jin1,2; Liu, Pingfang1,2; Zhao, Meiping3; Makrigiorgos, Gerasimos Mike1,2 |
| 刊名 | DIAGNOSTIC MOLECULAR PATHOLOGY
 |
| 出版日期 | 2011-06-01 |
| 卷号 | 20期号:2页码:81-89 |
| 关键词 | Cold-pcr Hrm Kras Lung Adenocarcinoma Mutation Detection |
| ISSN号 | 1052-9551 |
| DOI | 10.1097/PDM.0b013e3181fde92f |
| 英文摘要 | KRAS mutations exhibit significant predictive and prognostic value in cancer. Efficient, sensitive, and accurate molecular approaches are required to evaluate KRAS mutation status, even when mutant alleles are restricted to a small portion of a clinical sample, which otherwise contains wild-type alleles. We describe a highly sensitive method to detect KRAS mutations by high-resolution melting (HRM) analysis after mutation enrichment by fast-COLDpolymerase chain reaction (PCR). Using 10 ng of starting DNA and after fast-COLD-PCR of a 76-bp region containing KRAS codons 12 and 13; the amplicons undergo a nested conventional PCR reaction followed by HRM analysis. Samples exhibiting aberrant melting profiles are sequenced to identify mutation type and position. Serial dilution experiments indicate a sensitivity of approximately 0.3% mutant-to-wild type for HRM-based mutation detection and the ability to directly sequence mutation-containing samples. A number of lung adenocarcinoma specimens earlier characterized were screened. Fast-COLD-PCR-HRM analysis correctly identified KRAS mutations and also showed a previously undetected, low-level missense GGT > TTT complex mutation. On account of the short target regions and low requirement of starting DNA, this rapid, cost-effective, and sensitive fast-COLD-PCR-HRM approach is expected to find broad application for detecting low-abundance KRAS mutations in a wide range of clinical specimens. |
| 语种 | 英语 |
| WOS记录号 | WOS:000290662100003 |
| 出版者 | LIPPINCOTT WILLIAMS & WILKINS |
| 源URL | [http://ir.iccas.ac.cn/handle/121111/72479]  |
| 专题 | 中国科学院化学研究所 |
| 通讯作者 | Makrigiorgos, Gerasimos Mike |
| 作者单位 | 1.Harvard Univ, Brigham & Womens Hosp, WDivis Genome Stabil & DNA Repair, Dana Farber Brigham & Womens Canc Ctr,Sch Med, Boston, MA 02115 USA 2.Harvard Univ, Sch Med, Dept Radiat Oncol, Div Med Phys & Biophys, Boston, MA 02115 USA 3.Peking Univ, Coll Chem & Mol Engn, Minist Educ, BNLMS,Key Lab Bioorgan Chem & Mol Engn, Beijing 100871, Peoples R China |
| 推荐引用方式 GB/T 7714 | Song, Chen,Milbury, Coren A.,Li, Jin,et al. Rapid and Sensitive Detection of KRAS Mutation After fast-COLD-PCR Enrichment and High-resolution Melting Analysis[J]. DIAGNOSTIC MOLECULAR PATHOLOGY,2011,20(2):81-89. |
| APA | Song, Chen,Milbury, Coren A.,Li, Jin,Liu, Pingfang,Zhao, Meiping,&Makrigiorgos, Gerasimos Mike.(2011).Rapid and Sensitive Detection of KRAS Mutation After fast-COLD-PCR Enrichment and High-resolution Melting Analysis.DIAGNOSTIC MOLECULAR PATHOLOGY,20(2),81-89. |
| MLA | Song, Chen,et al."Rapid and Sensitive Detection of KRAS Mutation After fast-COLD-PCR Enrichment and High-resolution Melting Analysis".DIAGNOSTIC MOLECULAR PATHOLOGY 20.2(2011):81-89. |
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来源：化学研究所
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