title: Effect of specific or random c-DNA priming on sensitivity of tyrosinase nested RT-PCR: potential clinical relevance.
abstract: The reverse transcriptase polymerase chain reaction (RT-PCR) can be of clinical relevance in identifying malignant melanoma cells in blood or tissues of patients at risk for disseminated melanoma. The diagnostic value of this marker however, is still controversial. The objective of this study was to compare and quantify the difference in sensitivity of the nested RT-PCR for tyrosinase, with respect to the method utilized to produce the template c-DNA. We found a difference of a factor 10 in favor of a specific priming versus a random one. We concluded that this difference can be exploited in the analysis of blood samples. However, in the analysis of lymph node specimens, where the chance of positivity due to tyrosinase positive non-melanoma cells is much higher, the choice of a highly sensitive assay should be made with caution.
authors.affiliation: Department of Medical Oncology, University Hospital Groningen, The Netherlands.
authors.forename: A
authors.initials: A
authors.lastname: Calogero
authors.forename: G A
authors.initials: GA
authors.lastname: Hospers
authors.forename: H
authors.initials: H
authors.lastname: Timmer-Bosscha
authors.forename: H S
authors.initials: HS
authors.lastname: Koops
authors.forename: N H
authors.initials: NH
authors.lastname: Mulder
mesh_terms: D014408:Biomarkers, Tumor / Q000032:analysis* / Q000097:blood / Q000235:genetics; D017931:DNA Primers / Q000138:chemical synthesis* / Q000235:genetics; D018076:DNA, Complementary / Q000138:chemical synthesis* / Q000235:genetics; D006801:Humans; D008545:Melanoma / Q000097:blood / Q000201:enzymology; D014442:Monophenol Monooxygenase / Q000032:analysis* / Q000097:blood / Q000235:genetics; D009360:Neoplastic Cells, Circulating / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013698:Templates, Genetic; D014407:Tumor Cells, Cultured
chemical_list: D014408:Biomarkers, Tumor; D017931:DNA Primers; D018076:DNA, Complementary; D014442:Monophenol Monooxygenase