Identifying Patterns of Dna for Tumor Diagnosis Using Capillary Electrophoresis-amplified Fragment Length Polymorphism (ce-aflp) Screening

Amplified Fragment Length Polymorphism (AFLP) screening is a genome-wide genotyping strategy that has been widely used in plants and bacteria, but little has been reported concerning its use in humans. We investigated if the AFLP procedure could be coupled with high-throughput capillary electrophoresis (CE) for use in tumor diagnosis and classification. Using CE-AFLP, a series of molecular 'fingerprints' were generated for a set of gastric tumor and normal genomic DNA samples. The CE-AFLP procedure was qualitatively and quantitatively robust, and a variety of clustering tools were used to identify a specific DNA marker 'pattern' of 20 features that classified the tumor and normal samples to reasonable degrees of accuracy (Sensitivity 95%, Specificity 80%). The CE-AFLP-based approach also correctly classified 16 tumor samples, which in a previous study had exhibited no detectable genomic aberrations by comparative genome hybridization (CGH). This is the first reported application of CE-AFLP screening in tumor diagnosis. As the procedure is relatively inexpensive and requires minimal prior sequence knowledge and biological material, we suggest that CE-AFLP-based protocols may represent a promising new approach for DNA-based cancer screening and diagnosis.

[1]  P. Vos,et al.  AFLP: a new technique for DNA fingerprinting. , 1995, Nucleic acids research.

[2]  K. Singh,et al.  Identification of novel breast tumor-specific mutation(s) in the q11.2 region of chromosome 17 by RAPD/AP-PCR fingerprinting. , 2001, Gene.

[3]  Anne Kallioniemi,et al.  Targets of gene amplification and overexpression at 17q in gastric cancer. , 2002, Cancer research.

[4]  S. Lo,et al.  Prognostic Indicators for Survival After Curative Resection for Patients with Carcinoma of the Stomach , 1997, Digestive Diseases and Sciences.

[5]  B. Jahn,et al.  Comparative Evaluation of Three Different Genotyping Methods for Investigation of Nosocomial Outbreaks of Legionnaires' Disease in Hospitals , 2000, Journal of Clinical Microbiology.

[6]  F. Mitelman,et al.  Catalog of Chromosome Aberrations in Cancer , 1996, British Journal of Cancer.

[7]  K. Walder,et al.  AFLP fingerprinting of the human genome , 2000, Human Genetics.

[8]  J. Mesirov,et al.  Molecular classification of cancer: class discovery and class prediction by gene expression monitoring. , 1999, Science.

[9]  B Johansson,et al.  Chromosomal imbalance maps of malignant solid tumors: a cytogenetic survey of 3185 neoplasms. , 1997, Cancer research.

[10]  E. Petricoin,et al.  Use of proteomic patterns in serum to identify ovarian cancer , 2002, The Lancet.

[11]  D. Parkin,et al.  Case-control study of gastric cancer screening in Venezuela. , 1994, British Journal of Cancer.

[12]  N. Sampas,et al.  Molecular classification of cutaneous malignant melanoma by gene expression profiling , 2000, Nature.

[13]  D. Spandidos,et al.  p53 codon 72 polymorphism as a risk factor in the development of breast cancer. , 2000, Molecular cell biology research communications : MCBRC.

[14]  I. Tsuji,et al.  The evaluation of screening for gastric cancer in miyagi prefecture, Japan: A population‐based case‐control study , 1995, International journal of cancer.

[15]  Ash A. Alizadeh,et al.  Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling , 2000, Nature.