11/8/2022 0 Comments Semi quantitative pcrCategorical data were compared by the two-tailed Fisher exact test and the two-tailed Mann-Whitney U-test was used to compare median results from different assays. Duplicate blind assessments of MYCN copy number in the 101 tumor samples were performed by two independent observers.ĭata were analyzed using the SPSS software, version 16.0 (SPSS Inc., USA). After definition of the relative density, the copy number of the MYCN gene was calculated using the following equation: MYCN copy number = 1.76 x (relative intensity of the MYCN band) - 1.54 (7). Similar results were obtained for 100 (99%) samples, no significant difference was detected between the median log10 MYCN copy number (1.53 by SQ-PCR versus 1.55 by absolute real-time PCR), and the results of the two assays correlated closely (r = 0.8, Pearson correlation P 0.72 AU) were scored as amplified MYCN while products with intensity significantly lower (<0.36) than the references were scored as unamplified MYCN. To determine the usefulness of semi-quantitative differential PCR (SQ-PCR) for accurate quantification of MYCN gene copy number, we evaluated the analytical performance of this method by comparing the results obtained with it for 101 tumor samples of neuroblastoma to that obtained by absolute and relative real-time PCR. Bendit 1ġDisciplina de Hematologia e Hemoterapia, Hospital das Clínicas, 2Departamento de Endocrinologia e Metabologia, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, BrasilģFundação Pro-Sangue, Hemocentro, São Paulo, SP, BrasiĪmplification of the MYCN gene in neuroblastomas is a potent biological marker of highly aggressive tumors, which are invariably fatal unless sound clinical management is applied. The performance of semi-quantitative differential PCR is similar to that of real-time PCR for the detection of the MYCN gene in neuroblastomasĪ.C.M.F. Neuroblastoma Semi-quantitative PCR Real-time PCR MYCN Amplificationīraz J Med Biol Res, September 2009, Volume 42(9) 791-795 (Short Communication) Thus, SQ-PCR can be reliably used as an alternative assay in laboratories without facilities for real-time PCR. These findings indicate that the performance of SQ-PCR was comparable to that of real-time PCR for the amplification and quantification of MYCN copy number. In the comparison of SQ-PCR and relative real-time PCR, SQ-PCR versus relative real-time PCR concordant results were found in 100 (99%) samples, no significant difference was found in median log10 MYCN copy number (1.53 by SQ-PCR versus 1.27 by relative real-time PCR), and the results of the two assays correlated closely (r = 0.8, Pearson correlation P < 0.001). Similar results were obtained for 100 (99%) samples, no significant difference was detected between the median log10 MYCN copy number (1.53 by SQ-PCR versus 1.55 by absolute real-time PCR), and the results of the two assays correlated closely (r = 0.8, Pearson correlation P < 0.001). Amplification of the MYCN gene in neuroblastomas is a potent biological marker of highly aggressive tumors, which are invariably fatal unless sound clinical management is applied.
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