Application of multiplex FISH, CGH and MSSCP techniques for cytogenetic

Application of multiplex FISH, CGH and MSSCP techniques for cytogenetic

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Bladder cancer is one of the most common malig- nant cancers in Poland. Ninety percent of all blad- der carcinomas are transitional cell carcinomas (TCCs). TCC, with invasion limited to the lamina propria (pT1), causes serious clinical problems be- cause of 20–30% progression into invasive cancer, occurring in these tumours after transurethral re- section (TUR). Although cystoscopy is considered an invasive method, it belongs to a set of standard diagnostic procedures routinely applied in patients with clinical symptoms. Currently this method is also necessary to monitor patients for bladder can- cer recurrence and/or progression after treatment (Boman et al. 2002). We examined 25 patients with cystoscopically positive TCC, including 5 cases with pT1G1, 10 cases with pT2G3, and 10 cases with pT3G3 tu- mours, by using the multiplex FISH (UroVysion), Comparative Genomic Hybridization (CGH) and Multitemperature Single-Strand Conforma- tion Polymorphism (MSSCP) techniques. The UroVysion was performed according to the manufacturer’s instructions (VYSIS). The interphase nuclei were scored on the cell-by- cell basis in 2 steps. In the first step, 20 target cells were selected on the basis of large nuclear size or irregular nuclear shape. In a subsequent step, the FISH pattern was recorded in selected target cells. The result of UroVysion was identified as positive if 4 or more than 4 of 20 target cells showed polysomies of chromosomes 3 and 7 or 3 and 17 and/or 50% of the cells showed a loss of 9p21 (Figure 1A–C). The CGH and MSSCP techniques were per- formed according to the procedures described in detail elsewhere (Ka³u¿ewski et al. 2003; Phillips et al. 2000). J Appl Genet 47(3), 2006, pp. 273–275
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This note was uploaded on 02/22/2011 for the course HTEC 50 taught by Professor Hassel,patricia during the Spring '11 term at DeAnza College.

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Application of multiplex FISH, CGH and MSSCP techniques for cytogenetic

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