Quality Control Study on Melanoma Cell Detection in Blood
Author Information
Author(s): de Vries T J, Fourkour A, Punt C J A, van de Locht L T F, Wobbes T, Bosch S van den, Rooij M J M de, Mensink E J B M, Ruiter D J, Muijen G N P van
Primary Institution: University Hospital, Nijmegen, The Netherlands
Hypothesis
The study aimed to improve insight into the reproducibility of RT-PCR for detecting circulating melanoma cells.
Conclusion
The low reproducibility of the assay for detecting circulating melanoma cells is due to low numbers of amplifiable target mRNA molecules, not differences in mRNA quality.
Supporting Evidence
- 80% of blood samples were negative for tyrosinase.
- 66% of blood samples were negative for MART-1.
- Only four samples were positive in all four determinations for tyrosinase.
- Seven samples were positive in all four determinations for MART-1.
- MART-1 PCR had a better performance than tyrosinase PCR.
- Sensitivity increased when both markers were used.
Takeaway
Researchers checked blood samples from melanoma patients to see if they could find cancer cells. They found that it was hard to detect these cells because there weren't many of them in the blood.
Methodology
The study involved two reverse transcriptions on each mRNA sample and performed tyrosinase and MART-1 nested PCRs in duplicate per cDNA sample.
Limitations
The study's findings may be limited by the low number of positive results for melanoma markers.
Participant Demographics
68 melanoma patients, mainly stages III and IV.
Digital Object Identifier (DOI)
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