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Cell Biochemistry and Biophysics

, Volume 62, Issue 3, pp 415–420 | Cite as

Comparative Screening of K-ras Mutations in Colorectal Cancer and Lung Cancer Patients Using a Novel Real-Time PCR with ADx-K-ras Kit and Sanger DNA Sequencing

  • Haiping ZhangEmail author
  • Xiongwei Zheng
  • Tianhai Ji
  • Li Fu
  • Dongyu Bai
  • Yongqiang Liao
  • Haifang Zhang
  • Yi Ding
  • Limou Zheng
Original Paper

Abstract

We determined frequency/types of K-ras mutations in colorectal/lung cancer. ADx-K-ras kit (real-time/double-loop probe PCR) was used to detect somatic tumor gene mutations compared with Sanger DNA sequencing using 583 colorectal and 244 lung cancer paraffin-embedded clinical samples. Genomic DNA was used in both methods; mutation rates at codons 12/13 and frequency of each mutation were detected and compared. The data show that 91.4% colorectal and 59.0% lung carcinoma samples were detected conclusively by DNA sequencing, whereas 100% colorectal and lung samples were detected by ADx-K-ras kit. K-ras gene mutations were detected in 32.9–27.4% colorectal samples using kit and sequencing methods, respectively. Whereas 10.6–8.3% lung cancer samples were positively detected by kit and sequencing methods, respectively. Notably, 172/677 showed mutations and 467/677 showed wild type by both methods; 38 samples showed mutations with kit but wild type with sequencing. Mutations in colorectal samples were as follows: GGT → GAT/codon-12 (35.1%); GGC → GAC/codon-13 (26.6%); GGT → GTT/codon-12 (18.2%); and GGT → GCT/codon-12 (1.6%). Mutations in lung samples were as follows: GGT > GTT/codon-12 (40.9%) and GGT > GCT/codon-12 (4.5%). In conclusion, K-ras mutations involved 32.2% colorectal and 10.6% lung samples among this cohort. ADx-K-ras real-time PCR showed higher detection rates (P < 0.05). The kit method has good clinical applicability as it is simple, fast, less prone to contamination and hence can be used effectively and reliably for clinical screening of somatic tumor gene mutations.

Keywords

K-ras gene Somatic mutation Colorectal cancer Lung cancer Real-time PCR Double-loop probe DNA sequencing 

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Haiping Zhang
    • 1
    Email author
  • Xiongwei Zheng
    • 2
  • Tianhai Ji
    • 3
  • Li Fu
    • 1
  • Dongyu Bai
    • 1
  • Yongqiang Liao
    • 1
  • Haifang Zhang
    • 1
  • Yi Ding
    • 1
  • Limou Zheng
    • 4
  1. 1.Department of Pathology, First Hospital of XiamenXiamen UniversityXiamenChina
  2. 2.Department of PathologyFujian Provincial Tumor HospitalFuzhouChina
  3. 3.Department of Pathology174 Hospital of Chinese People’s Liberation ArmyXiamenChina
  4. 4.Institute for Biomedical ResearchXiamen UniversityXiamenChina

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