Semiautomated quantitative detection of loss of heterozygosity in the tumor suppressor gene p53.
The most frequently altered gene in diverse tumor types is the tumor suppressor gene p53. Typically, normal function is inactivated by point mutation of one allele and deletion of the other. Therefore, loss of heterozygosity (LOH) of intragenic polymorphic markers is a strong indication for p53 involvement in a cancerous lesion. This study shows that a highly polymorphic short tandem repeat (STR) within intron 1 of p53 is an excellent marker for quantitative evaluation of LOH in tumor samples, whose multicolor, fluorescently tagged PCR products are analyzed and quantitated on an automated DNA sequencer. The range of error was analyzed in detail. Discrete allelic profiles were obtained following amplification of DNA from microdissected cell samples of patients with urogenital tumors. By calculating qLOH, the relative allele ratio of a tumor compared with healthy tissue, a quantitative expression for the LOH is obtained. PCR-based tumor DNA typing using fluorescent STR primers and automated analysis provides an enhanced level of accuracy and sensitivity required for routine analysis in clinical practice, where large numbers of tumor samples have to be processed.
Hahn
M
M
Justus-Liebig-Universität, Giessen, Germany.
Matzen
S E
SE
Serth
J
J
Pingoud
A
A
D000483:Alleles; D001483:Base Sequence; D004247:DNA / Q000032:analysis / Q000097:blood; D017931:DNA Primers; D005456:Fluorescent Dyes; D005782:Gels; D017353:Gene Deletion; D016158:Genes, p53; D006579:Heterozygote; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D017186:Titrimetry; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*; D014565:Urogenital Neoplasms / Q000097:blood / Q000235:genetics
D017931:DNA Primers; D005456:Fluorescent Dyes; D005782:Gels; D016159:Tumor Suppressor Protein p53; D004247:DNA