A polymerase chain reaction-based microsatellite typing assay used for tumor cell line identification.

摘要: In this report we describe the application of a polymerase chain reaction (PCR)-based DNA typing assay to analysis tumor cell line identity. We have applied technique analyze four lines purchased from American Type Culture Collection (SK-OV-3, SK-BR-3, OVCAR, HeLa) and isolated ascites fluids ovarian cancer patients (YAOVBIX1, YAOVBIX3, OC194, OC346). assay, three polymorphic tetranucleotide microsatellite loci (GABARB1, TH01, HPRTB) were amplified DNAs in radioactive PCR-reactions. The products resolved polyacrylamide gels exposed film produce individual-specific patterns for five (HeLa, OC194). However, lines, SK-OV-3, YAOVBIX1, OC436 had identical "fingerprints" at all loci. probability that observed profile match could occur between randomly selected heterologous was calculated be 1.32 x 10(-13). On basis analysis, identified two independent cross-contamination events involving SK-OV-3 adenocarcinoma line. PCR-based is technically straightforward produces discrete allelic bands associated with known population frequencies, allowing unequivocal interpretation patterns.