Evaluation of the reliability of chromosomal imbalances detected by combined use of universal DNA amplification and comparative genomic hybridization.
Comparative genomic hybridization (CGH) analysis of microscopic tumor samples is allowed by universal DNA amplification using degenerate oligonucleotide primed-PCR (DOP-PCR). To evaluate the reliablity of DOP-PCR CGH, we performed DOP-PCR CGH and standard CGH in parallel using DNAs extracted from 10 malignant tumors of the hepatobiliary tract and pancreas. Similar results were obtained by both methods with a few exceptions, indicating that DOP-PCR CGH provides cytogenetic information equivalent to that obtained from standard CGH. We also investigated the sensitivity of DOP-PCR CGH using sequential dilutions of DNA from microdissected tumor cells. DOP-PCR using 100 to 800 pg of template DNA yielded successful CGH results. However, less than 50 pg of template DNA was not suitable because of the small amount of generated DNA. These findings suggest that DOP-PCR CGH is applicable for CGH analysis of tiny specimens which are too small for standard CGH. Accordingly, DOP-PCR CGH analysis may become a useful method in clinical laboratory examination.
Harada
T
T
Department of Pathology, Yamaguchi University School of Medicine, Ube, Yamaguchi 755-8505, Japan. tharada@po.cc.yamaguchi-u.ac.jp
Shiraishi
K
K
Kusano
N
N
Umayahara
K
K
Kondoh
S
S
Okita
K
K
Sasaki
K
K
D000368:Aged; D001650:Bile Duct Neoplasms / Q000235:genetics; D006528:Carcinoma, Hepatocellular / Q000235:genetics; D002869:Chromosome Aberrations; D017931:DNA Primers; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics*; D004067:Digestive System Neoplasms / Q000235:genetics*; D005260:Female; D005706:Gallbladder Neoplasms / Q000235:genetics; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics; D008297:Male; D008875:Middle Aged; D021141:Nucleic Acid Amplification Techniques; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010190:Pancreatic Neoplasms / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004273:DNA, Neoplasm