Real-time quantitative PCR for the measurement of MYCN amplification in human neuroblastoma with the TaqMan detection system.
Real-time quantitative PCR for the measurement of MYCN amplification in human neuroblastoma with the TaqMan detection system.
复制标题
使用 TaqMan 检测系统实时定量 PCR 测量人神经母细胞瘤中的 MYCN 扩增。
DOI：
10.1093/clinchem/45.11.1918
复制
发表时间：
1999
影响因子：
9.3
通讯作者：
C. Orlando
中科院分区：
文献类型：
--
作者：
C. C. Raggi;Maria Letizia Bagnoni;G. Tonini;M. Maggi;G. Vona;P. Pinzani;K. Mazzocco;B. Bernardi;M. Pazzagli;C. Orlando
BACKGROUND
Neuroblastoma is the most common extracranial malignant solid tumor in children under 5 years and is characterized by a wide clinical and biological heterogeneity, from spontaneously regressive forms to cancers with a rapid and fatal progression. MYCN oncogene amplification is considered the most important prognostic factor to evaluate survival and therapeutic choices in these patients.
METHODS
Here we present a new assay for rapid and accurate measurement of MYCN amplification, based on real-time quantitative PCR with the TaqMan(TM) reaction. The degree of MYCN amplification was derived from the ratio of the MYCN oncogene and the single-copy reference gene, beta-actin. The absolute abundance of these two genes in tumor sample DNA was obtained by extrapolation on external calibration curves generated with reference DNA.
RESULTS
We found a variable degree of MYCN amplification, from 2 to 29, in 26 of 49 (53%) neuroblastomas. These results were well correlated to those obtained with a competitive PCR assay in the same samples (r = 0. 987). MYCN amplification was associated mainly with advanced cancer stages, and the analysis of overall survival confirmed that the measurement of MYCN amplification is a predictor of patient outcome in neuroblastoma. Patients without MYCN amplification had a cumulative survival significantly higher than patients with low (<9; P = 0.02) and high (>/=9; P = 0.03) oncogene amplification.
CONCLUSION
The assay is rapid and reproducible and does not require any post-PCR analytical procedure.
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DOI：
--
发表时间：
1993-05
期刊：
The American journal of pathology
影响因子：
--
作者：
D. Shapiro;tt Marcus B. Valentine;S. Rowe;A. E. Sinclair;J. Sublett;W. Roberts;A. Look
通讯作者：
D. Shapiro;tt Marcus B. Valentine;S. Rowe;A. E. Sinclair;J. Sublett;W. Roberts;A. Look
影响因子：
158.5
作者：
CHAN, HSL;HADDAD, G;LING, V
通讯作者：
LING, V
影响因子：
158.5
作者：
SEEGER, RC;BRODEUR, GM;HAMMOND, D
通讯作者：
HAMMOND, D
影响因子：
56.9
作者：
BRODEUR, GM;SEEGER, RC;BISHOP, JM
通讯作者：
BISHOP, JM