Development of fluorescent quantitation PCR method for detection of urinary CK20 / 重庆医科大学学报
Journal of Chongqing Medical University
;
(12)2003.
Artículo
en Chino
| WPRIM
| ID: wpr-579765
ABSTRACT
Objective:
To develop a novel real-time fluorescence PCR method using Taqman probe for convenient,fast,sensitive and specific detection of urinary CK20 and use in early diagnosis and follow-up of transitional cell carcinoma of the bladder(TCCB).Methods:
CK20 gene from cultured T24 cells was amplified by conventional RT-PCR,the standard quantitative plasmid was constructed by T-Aclone method.Taqman probe and primers was designed according to the sequence of CK20 cloned gene,the real-time PCR method for determination of CK20 mRNA was established and evaluated.Results:
The developed real-time PCR method showed high sensitivity(10 2copies /?l) and good specificity;the linear range was 102 ~109 copies /?l,the coefficient variation(CV) was 0.89 % in intra-assay and 2.45% in day to day.Conclusion:
A novel real-time fluorescence quantitative PCR method for detection of CK20 was established.It provided a basis for developing new diagnostic kits using the Taqman probe for clinical application.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Tipo de estudio:
Estudio diagnóstico
/
Estudio de tamizaje
Idioma:
Chino
Revista:
Journal of Chongqing Medical University
Año:
2003
Tipo del documento:
Artículo
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