Detection of Ki-ras mutations in tissue and plasma samples of patients with pancreatic cancer using PNA-mediated PCR clamping and hybridisation probes.
Detection of Ki-ras mutations in tissue and plasma samples of patients with pancreatic cancer using PNA-mediated PCR clamping and hybridisation probes.
复制标题
DOI：
10.1038/sj.bjc.6602319
复制
发表时间：
2005-01-31
影响因子：
8.8
通讯作者：
中科院分区：
文献类型：
--
作者：
在本研究中，我们将pcr -箝位法与使用突变特异性杂交探针和野生型特异性肽核酸（PNAs）的熔化曲线分析相结合，以确定胰腺癌患者组织和血浆样本中原癌基因Ki-ras密码子12最常见的点突变的基因型。灵敏度为1-5 × 10−5。4例患者组织样本的熔化曲线分析显示2个缬氨酸突变，1个无缬氨酸突变和1个野生型序列。在10例胰腺导管腺癌患者的非相关血浆样本中，28%的dna（64例中的18例）发现Ki-ras改变。缬氨酸突变是主要检测到的基因改变（83%）。在调查的10例患者中，4例患者（40%）在临床观察中出现Ki-ras突变阳性。所有4例患者均表现出疾病进展和高水平的肿瘤标志物CA 19-9。总之，所描述的一步程序可能是分析组织和血浆样本中Ki-ras点突变的有用临床工具。此外，该方法可适用于多个突变的同时检测和定量。
In the present study, we combined the PCR-clamping approach with melting curve analysis using mutant specific hybridisation probes and wild-type specific peptide nucleic acids (PNAs) to determine the genotypes of the most frequent point mutation in codon 12 of the proto-oncogene Ki-ras in tissue and plasma samples of patients with pancreatic cancer. The sensitivity of our assay was 1–5 × 10−5. The melting curve analysis of tissue samples of four patients revealed two valine mutations, one none-valine mutation and one wild-type sequence. Ki-ras alterations were found in 28% of DNAs (18 out of 64) of nonrelated plasma samples of 10 patients with ductal adenocarcinoma of the pancreas. The valine mutation was the predominantly detected gene alteration (83%). Out of ten patients investigated, four patients (40%) became positive during clinical observation with respect to Ki-ras mutation. All four patients exhibited progressive disease and high levels of tumour marker CA 19-9. In conclusion, the one-step procedure discribed may be a useful clinical tool for analysing Ki-ras point mutations in tissue and plasmas samples. In addition, this method can be adapted for simultanous detection of multiple mutations and quantitation.
登录
查看更多内容
影响因子：
--
作者：
Rhodes, CH;Honsinger, C;Sorenson, GD
通讯作者：
Sorenson, GD
影响因子：
50.5
作者：
Lamerz, R
通讯作者：
Lamerz, R
影响因子：
50.5
作者：
Goggins, M;Kern, SE;Hruban, RH
通讯作者：
Hruban, RH
影响因子：
3.6
作者：
Shi, X;Friess, H;Büchler, MW
通讯作者：
Büchler, MW
DOI：
10.1016/s1385-299x(01)00123-4
发表时间：
2002-02-01
期刊：
BRAIN RESEARCH PROTOCOLS
影响因子：
--
作者：
Ghanima, A;Bennis, M;Rampin, O
通讯作者：
Rampin, O