TY - JOUR
T1 - Comparison between three molecular methods for detection of blood melanoma tyrosinase mRNA. Correlation with melanoma stages and S100B, LDH, NSE biochemical markers.
AU - Santocito, Concetta
AU - Concolino, Paola
AU - Lavieri, Maria Michela
AU - Ameglio, Franco
AU - Gentileschi, Stefano
AU - Capizzi, Rodolfo
AU - Rocchetti, Sandro
AU - Amerio, Pierluigi
AU - Castagnola, Massimo
AU - Zuppi, Cecilia
AU - Capoluongo, Ettore Domenico
PY - 2005
Y1 - 2005
N2 - Abstract
BACKGROUND:
The molecular monitoring of circulating tumor cells by reverse transcriptase-PCR (RT-PCR) for patients with melanoma, is still under debate. It may be affected by: a) pre-analytical variability, b) frequency of melanoma-associated gene transcripts and c) the reliability of the methods employed. Few commercial methods are available for the detection of tyrosinase mRNA in blood.
OBJECTIVE:
Comparison between two RT-PCR-nested methods with a third one based on real-time methodology, for detection and quantitation of tyrosinase transcripts, respectively.
METHODS:
Sixty-two melanoma patients with different AJCC stages and 20 healthy subjects were enrolled. All blood samples were extracted in duplicate with two different methods. Two nested-PCR methods (one commercial and one in house) plus a real time commercial kit were employed.
RESULTS:
The two nested PCR methods employed were overimposable, specific and sensitive, at least in the stage III, where there was a concordance between sentinel lymph nodes detection and blood tyrosinase positivity. The different extraction methods did not affect the quality of results, while the commercial real-time kit cannot be used.
CONCLUSION:
Tyrosinase mRNA detection may be therefore employed to monitor the melanoma patients over time in function of response to therapy.
AB - Abstract
BACKGROUND:
The molecular monitoring of circulating tumor cells by reverse transcriptase-PCR (RT-PCR) for patients with melanoma, is still under debate. It may be affected by: a) pre-analytical variability, b) frequency of melanoma-associated gene transcripts and c) the reliability of the methods employed. Few commercial methods are available for the detection of tyrosinase mRNA in blood.
OBJECTIVE:
Comparison between two RT-PCR-nested methods with a third one based on real-time methodology, for detection and quantitation of tyrosinase transcripts, respectively.
METHODS:
Sixty-two melanoma patients with different AJCC stages and 20 healthy subjects were enrolled. All blood samples were extracted in duplicate with two different methods. Two nested-PCR methods (one commercial and one in house) plus a real time commercial kit were employed.
RESULTS:
The two nested PCR methods employed were overimposable, specific and sensitive, at least in the stage III, where there was a concordance between sentinel lymph nodes detection and blood tyrosinase positivity. The different extraction methods did not affect the quality of results, while the commercial real-time kit cannot be used.
CONCLUSION:
Tyrosinase mRNA detection may be therefore employed to monitor the melanoma patients over time in function of response to therapy.
KW - melanoma
KW - polymerase chain reaction
KW - s100
KW - tyrosinase
KW - melanoma
KW - polymerase chain reaction
KW - s100
KW - tyrosinase
UR - http://hdl.handle.net/10807/12574
M3 - Article
SN - 0009-8981
SP - 85
EP - 93
JO - Clinica Chimica Acta
JF - Clinica Chimica Acta
ER -