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AlinaPokhilko

§¯§Ñ§Ó§é§Ñ§Ý§î§ß§Ú§Û §Ü§å§â§ã: ?§®§à§Ý§Ö§Ü§å§Ý§ñ§â§ß§Ñ §à§ß§Ü§à§Ý§à§Ô?§ñ, §á§Ñ§ä§à§Ý§à§Ô?§ñ §ä§Ñ §Ô§Ö§ß§Ö§ä§Ú§Ü§Ñ? §®§Ö§Õ§Ú§é§ß§Ñ §Ý§Ñ§Ò§à§â§Ñ§ä§à§â?§ñ CSD §ã§á?§Ý§î§ß§à §Ù Center for research and education of translational biology and medicine (www.tbm.center ) §Ù§Ñ§á§å§ã§Ü§Ñ? §Ò§Ö§Ù§Ü§à§ê§ä§à§Ó§ß§Ú§Û §Ü§å§â§ã §ß§Ñ§Ó§é§Ñ§Ý§î§ß§Ú§ç §Ý§Ö§Ü§è?§Û §Õ§Ý§ñ §å§ã?§ç §Ò§Ñ§Ø§Ñ§ð§é§Ú§ç. §¡§ß§Ñ§ã§ä§Ñ§ã?§ñ §¶?§ä§î§à - §Ò?§à§Ý§à§Ô §Ó?§Õ§Õ?§Ý§å ?§Þ§å§ß§à§Ô?§ã§ä§à§ç?§Þ??, §ß§Ñ§á§â§ñ§Þ§Ü§å FISH §Õ§à§ã§Ý?§Õ§Ø§Ö§ß§ß§ñ

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§°§ã§ß§à§Ó§Ú §Þ§Ö§ä§à§Õ§å FISH, §Õ§à§ã§Ó?§Õ §Ó§Ú§Ü§à§â§Ú§ã§ä§Ñ§ß§ß§ñ §Ó §Ý§Ñ§Ò§à§â§Ñ§ä§à§â?? CSD §¢?§à§Ý§à§Ô §¶?§ä§î§à §¡§ß§Ñ§ã§ä§Ñ§ã?§ñ §¬§Ú?§Ó ¨C 2021
FISH - Fluorescence in situ Hybridization §¤?§Ò§â§Ú§Õ§Ú§Ù§Ñ§è?§ñ in situ ¨C §Ò§Ñ§Ù§à§Ó§Ú§Û §Þ§Ö§ä§à§Õ §Þ§à§Ý§Ö§Ü§å§Ý§ñ§â§ß§à-§è§Ú§ä§à§Ô§Ö§ß§Ö§ä§Ú§é§ß§Ú§ç §Õ§à§ã§Ý?§Õ§Ø§Ö§ß§î. ¡ñ §Õ§à§Ù§Ó§à§Ý§ñ? §Ó§Ú§ñ§Ó§Ú§ä§Ú §ã§á§Ö§è§Ú§æ?§é§ß§å §á§à§ã§Ý?§Õ§à§Ó§ß?§ã§ä§î §ß§å§Ü§Ý§Ö?§ß§à§Ó§à? §Ü§Ú§ã§Ý§à§ä§Ú (§¥§¯§¬ §é§Ú §²§¯§¬) §Ó §Ü§Ý?§ä§Ú§ß?. ¡ñ §±§â§Ú§ß§è§Ú§á §Þ§Ö§ä§à§Õ§å §á§à§Ý§ñ§Ô§Ñ? §å §Ü§à§Þ§á§Ý§Ö§Þ§Ö§ß§ä§Ñ§â§ß§à§Þ§å §Ù§Ó¡¯§ñ§Ù§å§Ó§Ñ§ß§ß? §ß§å§Ü§Ý§Ö§à§ä§Ú§Õ§ß§à§Ô§à §Ù§à§ß§Õ§å (§á§â§à§Ò§Ú) ?§Ù §á§à§ã§Ý?§Õ§à§Ó§ß?§ã§ä§ð §¥§¯§¬ §é§Ú §²§¯§¬ §Õ§à§ã§Ý?§Õ§Ø§å§Ó§Ñ§ß§à§Ô§à §Ù§â§Ñ§Ù§Ü§Ñ. §¤?§Ò§â§Ú§Õ§Ú§Ù§Ñ§è?§ñ in situ §¶§à§â§Þ§å§Ó§Ñ§ß§ß§ñ §Ô?§Ò§â§Ú§Õ?§Ó (§¥§¯§¬-§¥§¯§¬, §¥§¯§¬-§²§¯§¬, §²§¯§¬-§²§¯§¬) §£ §å§Þ§à§Ó§Ñ§ç §æ?§Ü§ã§à§Ó§Ñ§ß§à§Ô§à §ã§å§Ò§ã§ä§â§Ñ§ä§å, §Ó §Ü§Ý?§ä§Ú§ß? §¥§¯§¬ §Ù§â§Ñ§Ù§Ü§Ñ §©§à§ß§Õ
§¬§Ý§Ñ§ã§Ú§æ?§Ü§Ñ§è?§ñ §Ù§à§ß§Õ?§Ó ¡ñ §©§à§ß§Õ (§á§â§à§Ò§Ñ) ¨C §è§Ö §Ü§à§â§à§ä§Ü§Ñ §ß§å§Ü§Ý§Ö§à§ä§Ú§Õ§ß§Ñ §á§à§ã§Ý?§Õ§à§Ó§ß?§ã§ä§î (§à§á§ä§Ú§Þ§Ñ§Ý§î§ß§à 50-300 §á§Ñ§â §à§ã§ß§à§Ó), §Ü§à§Þ§á§Ý§Ö§Þ§Ö§ß§ä§Ñ§â§ß§Ñ §Õ§à§ã§Ý?§Õ§Ø§å§Ó§Ñ§ß?§Û §á§à§ã§Ý?§Õ§à§Ó§ß§à§ã§ä? §ß§å§Ü§Ý§Ö?§ß§à§Ó§à? §Ü§Ú§ã§Ý§à§ä§Ú. §©§Ñ §Þ?§ã§è§Ö§Þ §á§â§Ú?§Õ§ß§Ñ§ß§ß§ñ: a) §Ý§à§Ü§å§ã§ã§á§Ö§è§Ú§æ?§é§ß? (LSI - Locus Specific Identifier) b) §è§Ö§ß§ä§â§à§Þ§Ö§â§ß? (CEP) c) §ä§Ö§Ý§à§Þ§Ö§â§ß? d) §Õ§à §Ó§ã??? §ç§â§à§Þ§à§ã§à§Þ§Ú (WCP - Whole chromosome painting) §©§Ñ §Ò§å§Õ§à§Ó§à§ð: a) §Õ§Ó§à§Ý§Ñ§ß§è§ð§Ô§à§Ó? §¥§¯§¬-§Ù§à§ß§Õ§Ú, b) §à§Õ§ß§à§Ý§Ñ§ß§è§ð§Ô§à§Ó? §Ñ§ß§ä§Ú§ã§Ö§ß§ã-§²§¯§¬-§Ù§à§ß§Õ§Ú (§â§Ú§Ò§à§á§â§à§Ò§Ú), c) §à§Õ§ß§à§Ý§Ñ§ß§è§ð§Ô§à§Ó? §¥§¯§¬-§Ù§à§ß§Õ§Ú, d) §ã§Ú§ß§ä§Ö§ä§Ú§é§ß? §à§Ý?§Ô§à§Õ§Ö§Ù§à§Ü§ã§Ú§ß§å§Ü§Ý§Ö§à§ä§Ú§Õ§ß? §Ù§à§ß§Õ§Ú §ä§Ñ §à§Ý?§Ô§à§â§Ú§Ò§à§Ù§à§ß§Õ§Ú
§©§Ñ §ã§á§à§ã§à§Ò§à§Þ §Þ?§é§Ö§ß§ß§ñ: §©§à§ß§Õ§Ú §© §â§Ñ§Õ?§à§Ñ§Ü§ä§Ú§Ó§ß§à§ð §Þ?§ä§Ü§à§ð §© §ß§Ö§â§Ñ§Õ?§à§Ñ§Ü§ä§Ú§Ó§ß§à§ð §Þ?§ä§Ü§à§ð P32, S35, H3 §¢?§à§ä§Ú§ß §¥§Ú§Ô§à§Ü§ã§Ú§Ô§Ö§ß?§ß (DIG) §¶§Ý§å§à§â§à§ç§â§à§Þ §Þ?§ä§Ü§Ú fluorescein rhodamine coumarin Texas Red SpectrumOrange SpectrumGreen
§£§Ú§Õ§Ú §Ô?§Ò§â§Ú§Õ§Ú§Ù§Ñ§è?? in situ ISH §²§Ñ§Õ?§à§Ñ§Ü§ä§Ú§Ó§ß§Ú§Û (§Ñ§Ó§ä§à§â§Ñ§Õ?§à§Ô§â§Ñ§æ?§ñ) BRISH Bright field in situ hybridization (§ã§Ó?§ä§Ý§à§Ó§Ñ §Þ?§Ü§â§à§ã§Ü§à§á?§ñ) §³ISH Chromogenic in situ hybridization SISH Silver in situ hybridization ?§ß§ê? FISH Fluorescent in situ hybridization (§æ§Ý§å§à§â§Ö§ã§è§Ö§ß§ä§ß§Ñ §Þ?§Ü§â§à§ã§Ü§à§á?§ñ) Rainbow-FISH Raman-FISH ReD-FISH Reverse-FISH RING-FISH RNA-FISH RxFISH Split-Signal FISH T-FISH 3-D FISH Zoo-FISH ACM-FISH armFISH CARD-FISH catFISH CB-FISH CO-FISH COBRA-FISH COD-FISH COMBO-FISH Comet-FISH Cryo-FISH D-FISH DBD-FISH e-FISH Fiber-FISH Flow-FISH Fusion-Signal FISH Halo-FISH Harlequin-FISH Immuno-FISH LNA-FISH M-FISH Multilocus or ML-FISH PCC-FISH PNA-FISH Q-FISH QD-FISH
§±§â§Ú§ß§è§Ú§á §ä§Ñ §à§ã§ß§à§Ó§ß? §Ö§ä§Ñ§á§Ú §Þ§Ö§ä§à§Õ§å FISH/CISH (§ß§Ñ §á§â§Ú§Ü§Ý§Ñ§Õ? §Ô?§ã§ä§à§Ý§à§Ô?§é§ß§à§Ô§à §Þ§Ñ§ä§Ö§â?§Ñ§Ý§å) §±§Ñ§â§Ñ§æ?§ß§à§Ó§Ú§Û §Ò§Ý§à§Ü §Ù §æ?§Ü§ã§à§Ó§Ñ§ß§à§ð §å §æ§à§â§Þ§Ñ§Ý?§ß? §ä§Ü§Ñ§ß§Ú§ß§à§ð §£§Ú§Ô§à§ä§à§Ó§Ý§Ö§ß§ß§ñ §Ù§â?§Ù?§Ó §ä§à§Ó§ë§Ú§ß§à§ð 2-3 ¦Ìm §¥§Ö§á§Ñ§â§Ñ§æ?§ß?§Ù§Ñ§è?§ñ §å §Ü§ã§Ú§Ý§à§Ý? §¥§Ö§Ô?§Õ§â§Ñ§ä§Ñ§è?§ñ §å §ã§Ö§â?? §ã§á§Ú§â§ä?§Ó §Ó §á§à§â§ñ§Õ§Ü§å §Ù§Þ§Ö§ß§ê§Ö§ß§ß§ñ ?§ç §Ü§à§ß§è§Ö§ß§ä§â§Ñ§è?? §¶§Ö§â§Þ§Ö§ß§ä§Ñ§ä§Ú§Ó§ß§Ú§Û §á§â§à§ä§Ö§à§Ý?§Ù (§ß§Ñ§Û§é§Ñ§ã§ä?§ê§Ö §Ù §Ó§Ú§Ü§à§â§Ú§ã§ä§Ñ§ß§ß§ñ§Þ §á§Ö§á§ã§Ú§ß§å) §¥§à§Õ§Ñ§Ó§Ñ§ß§ß§ñ §Þ?§é§Ö§ß§à? §á§â§à§Ò§Ú §¥§Ö§ß§Ñ§ä§å§â§Ñ§è?§ñ (82 ¡ã§³, 5 §ç§Ó) §¤?§Ò§â§Ú§Õ§Ú§Ù§Ñ§è?§ñ 37 ¡ã§³ (18 §Ô§à§Õ) §±§à§ã§ä§Ô?§Ò§â§Ú§Õ§Ú§Ù§Ñ§è?§Û§ß§Ñ §Ó?§Õ§Þ§Ú§Ó§Ü§Ñ §¥§Ö§ä§Ö§Ü§è?§ñ §±§â§ñ§Þ§Ñ (§á§â§à§Ò§Ñ, §Þ?§é§Ö§ß§Ñ §æ§Ý§å§à§â§à§ç§â§à§Þ§à§Þ, FISH §Þ§Ö§ä§à§Õ) §¯§Ö§á§â§ñ§Þ§Ñ (§á§â§à§Ò§Ñ, §Þ?§é§Ö§ß§Ñ §Ò?§à§ä§Ú§ß§à§Þ §Ñ§Ò§à §Õ§Ú§Ô§à§Ü§ã§Ú§Ô§Ö§ß?§ß§à§Þ, §³ISH §Þ§Ö§ä§à§Õ) §¬§à§ß§ä§â§æ§Ñ§â§Ò§å§Ó§Ñ§ß§ß§ñ §Ù 4¡¯,6-§Õ?§Ñ§Þ?§ß§à-2- §æ§Ö§ß?§Ý?§ß§Õ§à§Ý§à§Þ (DAPI)
§±§â§Ú§ß§è§Ú§á §Þ§Ö§ä§à§Õ§å CISH HRP ¨C Horseradish peroxidase (§á§Ö§â§à§Ü§ã§Ú§Õ§Ñ§Ù§Ñ §ç§â§à§ß§å) DAB ¨C Diaminobenzidine ¨C §ã§å§Ò§ã§ä§â§Ñ§ä §Õ§Ý§ñ HRP EBV (CISH)
¡ñ §±§â§à§Ó§à§Õ§Ú§ä§î§ã§ñ §Ù§Ñ §Õ§à§á§à§Þ§à§Ô§à§ð §æ§Ý§å§à§â§Ö§ã§è§Ö§ß§ä§ß§à§Ô§à §Þ?§Ü§â§à§ã§Ü§à§á§Ñ. ¡ñ §¡§ß§Ñ§Ý?§Ù ? §á?§Õ§â§Ñ§ç§å§ß§à§Ü §Ü§Ý?§ä§Ú§ß §Ó?§Õ§Ò§å§Ó§Ñ?§ä§ã§î§ñ §á§â§Ú §Ó§Ö§Ý§Ú§Ü§à§Þ§å §Ù§Ò?§Ý§î§ê§Ö§ß§ß? (10§ç100) §Ù §Ó§Ú§Ü§à§â§Ú§ã§ä§Ñ§ß§ß§ñ§Þ ?§Þ§Ö§â§ã?§Û§ß§à? §à§Ý?? §Õ§Ý§ñ §æ§Ý§å§à§â§Ö§ã§è§Ö§ß§ä§ß§à§Ô§à §Þ?§Ü§â§à§ã§Ü§à§á§Ñ (Type FF) ?§ß§ä§Ö§â§á§â§Ö§ä§Ñ§è?§ñ §â§Ö§Ù§å§Ý§î§ä§Ñ§ä?§Ó FISH DAPI Green/orange §æ?§Ý§î§ä§â
a) Single color probe b) Dual color probe c) Multi color probe d) Dual color Break apart probe e) Dual color Fusion probe Dual Color Probe Multi color probe Dual color Fusion probe Dual Color Break Apart Probe §°§á§Ú§ã (§Ü§Ñ§â§ä§Ñ) §á§â§à§Ò§Ú
§±§â§Ñ§Ü§ä§Ú§é§ß§Ö §Ù§Ñ§ã§ä§à§ã§å§Ó§Ñ§ß§ß§ñ FISH (CISH) §Ó §Þ§Ö§Õ§Ú§è§Ú§ß? ¡ñ §£§Ú§ñ§Ó§Ý§Ö§ß§ß§ñ §Ó?§â§å§ã§ß§Ú§ç §Ô§Ö§ß§à§Þ?§Ó (EBV, HPV) ¡ñ §£§Ú§ñ§Ó§Ý§Ö§ß§ß§ñ §Ô§Ö§ß§Ö§ä§Ú§é§ß§Ú§ç §á§à§â§å§ê§Ö§ß§î §ß§Ñ §Þ§Ö§ä§Ñ§æ§Ñ§Ù§ß§Ú§ç §ç§â§à§Þ§à§ã§à§Þ§Ñ§ç §Ñ§Ò§à §Ó ?§ß§ä§Ö§â§æ§Ñ§Ù§ß§Ú§ç §ñ§Õ§â§Ñ§ç §Ü§Ý?§ä§Ú§ß (§ç§â§à§Þ§à§ã§à§Þ§ß? §Ñ§Ò§Ö§â§Ñ§è??, §ä§â§Ñ§ß§ã§Ý§à§Ü§Ñ§è??, §Õ§Ö§Ý§Ö§è??, §Ñ§Þ§á§Ý?§æ?§Ü§Ñ§è?? §ä§à§ë§à) ¡ñ §¥§Ú§æ§Ö§â§Ö§ß§è?§Û§ß§Ñ §Õ?§Ñ§Ô§ß§à§ã§ä§Ú§Ü§Ñ §Ó §à§ß§Ü§à§Ý§à§Ô?? ¡ñ §£§Ú§Ù§ß§Ñ§é§Ö§ß§ß§ñ §é§å§ä§Ý§Ú§Ó§à§ã§ä? §Õ§à §ä§Ñ§â§Ô§Ö§ä§ß§Ú§ç §á§â§Ö§á§Ñ§â§Ñ§ä?§Ó §Ó §à§ß§Ü§à§Ý§à§Ô?? ¡ñ §£§Ú§Ù§ß§Ñ§é§Ö§ß§ß§ñ §Ô§â§å§á §â§Ú§Ù§Ú§Ü§å §Ó §à§ß§Ü§à§Ý§à§Ô?? ¡ñ §±§â§Ö§ß§Ñ§ä§Ñ§Ý§î§ß§Ñ §Õ?§Ñ§Ô§ß§à§ã§ä§Ú§Ü§Ñ
§±§â§Ú§Ü§Ý§Ñ§Õ§Ú §Ù§Ñ§ã§ä§à§ã§å§Ó§Ñ§ß§ß§ñ §Þ§Ö§ä§à§Õ§å FISH §Ó §Ý§Ñ§Ò§à§â§Ñ§ä§à§â?? CSD ? §£ 1960 §Ô Peter Nowell §ä§Ñ David Hungerford §à§á§Ú§ã§Ñ§Ý§Ú §ã§á§Ö§è§Ú§æ?§é§ß§å §ç§â§à§Þ§à§ã§à§Þ§å, §ñ§Ü§å §ß§Ñ§Ù§Ó§Ñ§Ý§Ú §æ?§Ý§Ñ§Õ§Ö§Ý§î§æ?§Û§ã§î§Ü§à§ð. ? §¶?§Ý§Ñ§Õ§Ö§Ý§î§æ?§Û§ã§î§Ü§Ñ §ç§â§à§Þ§à§ã§à§Þ§Ñ (Ph.) ¨C §è§Ö §â§Ö§Ù§å§Ý§î§ä§Ñ§ä §â§Ö§è§Ú§á§â§à§Ü§ß§à? §ä§â§Ñ§ß§ã§Ý§à§Ü§Ñ§è??* §Þ?§Ø §Õ?§Ý§ñ§ß§Ü§Ñ§Þ§Ú §Õ§à§Ó§Ô§à§Ô§à §á§Ý§Ö§é§Ñ 22 (§Õ?§Ý§ñ§ß§Ü§Ñ 22q11.23, §Ô§Ö§ß BCR) §ä§Ñ 9 (§Õ?§Ý§ñ§ß§Ü§Ñ 9q34.12, §Ô§Ö§ß ABL1) §ç§â§à§Þ§à§ã§à§Þ§Ú. ? §µ 90% §á§Ñ§è??§ß§ä?§Ó §Ù §ç§â§à§ß?§é§ß§Ú§Þ §Þ??§Ý§à§Ý§Ö§Û§Ü§à§Ù§à§Þ ? 25% §Ù §Ô§à§ã§ä§â§Ú§Þ §Ý?§Þ§æ§à§Ò§Ý§Ñ§ã§ä§ß§Ú§Þ §Ý§Ö§Û§Ü§à§Ù§à§Þ §ß§Ñ§ñ§Ó§ß§Ñ §ä§â§Ñ§ß§ã§Ý§à§Ü§Ñ§è?§ñ ? BCR/ABL t (9; 22) (q34.1; q11.2). *§²§Ö§è§Ú§á§â§à§Ü§ß§Ñ §ä§â§Ñ§ß§ã§Ý§à§Ü§Ñ§è?§ñ - §à§Ò§Þ?§ß §Õ?§Ý§ñ§ß§Ü§Ñ§Þ§Ú §Þ?§Ø §Õ§Ó§à§Þ§Ñ §ß§Ö§Ô§à§Þ§à§Ý§à§Ô?§é§ß§Ú§Þ§Ú §ç§â§à§Þ§à§ã§à§Þ§Ñ§Þ§Ú §£§Ú§Ù§ß§Ñ§é§Ö§ß§ß§ñ ?§¶?§Ý§Ñ§Õ§Ö§Ý§î§æ?§Û§ã§î§Ü§à? §ç§â§à§Þ§à§ã§à§Þ§Ú? (BCR/ABL, t (9; 22) (q34.1; q11.2))
BCR/ABL - (2R2G) BCR/ABL + (2F1R1G) BCR/ABL + (1F1R1G) §®§Ñ§ä§Ö§â?§Ñ§Ý: §Ü§â§à§Ó §Ñ§Ò§à §Ñ§ã§á?§â§Ñ§ä §Ü?§ã§ä§Ü§à§Ó§à§Ô§à §Þ§à§Ù§Ü§å §±?§Õ§â§Ñ§ç§å§ß§à§Ü §Þ?§ß?§Þ§å§Þ 100 §Ü§Ý?§ä§Ú§ß Cut off ¨C 2 % §±§â§à§Ò§Ñ: Dual color Fusion probe §£§Ú§Ù§ß§Ñ§é§Ö§ß§ß§ñ §æ?§Ý§Ñ§Õ§Ö§Ý§î§æ?§Û§ã§î§Ü§à? §ç§â§à§Þ§à§ã§à§Þ§Ú BCR/ABL
§£§Ú§Ù§ß§Ñ§é§Ö§ß§ß§ñ §é§å§ä§Ý§Ú§Ó§à§ã§ä? §Õ§à §ä§Ñ§â§Ô§Ö§ä§ß§Ú§ç §á§â§Ö§á§Ñ§â§Ñ§ä?§Ó §¥§à§ã§Ý?§Õ§Ø§Ö§ß§ß§ñ §Ñ§Þ§á§Ý?§æ?§Ü§Ñ§è?? §Ô§Ö§ß§Ñ Her2/neu (§â§Ñ§Ü §Þ§à§Ý§à§é§ß§à? §Ù§Ñ§Ý§à§Ù§Ú, §â§Ñ§Ü §ê§Ý§å§ß§Ü§Ñ, §Ü§à§Ý§à§â§Ö§Ü§ä§Ñ§Ý§î§ß§Ú§Û §â§Ñ§Ü) ? §¤§Ö§ß ERBB2 (Her2/neu) §â§à§Ù§ä§Ñ§ê§à§Ó§Ñ§ß§Ú§Û §ß§Ñ §Õ§à§Ó§Ô§à§Þ§å §á§Ý§Ö§é? 17 §ç§â§à§Þ§à§ã§à§Þ§Ú (17q12) ? §¤§Ö§ß ERBB2 §Ü§à§Õ§å? §Ò?§Ý§à§Ü 185-190 §Ü§¥§Ñ (Her2/neu), §ë§à ? §â§Ö§è§Ö§á§ä§à§â§à§Þ §Ù §ä§Ú§â§à§Ù§Ú§ß§Ü?§ß§Ñ§Ù§ß§à§ð §Ñ§Ü§ä§Ú§Ó§ß?§ã§ä§ð §ä§Ñ §Ò§Ö§â§Ö §å§é§Ñ§ã§ä§î §Ó §â§Ö§Ô§å§Ý§ñ§è?? §Ü§Ý?§ä§Ú§ß§ß§à§Ô§à §â§à§ã§ä§å. ? §¢?§Ý§à§Ü Her2/neu ? §Þ?§ê§Ö§ß§ß§ð §Õ§Ý§ñ §ä§Ñ§â§Ô§Ö§ä§ß§Ú§ç §á§â§Ö§á§Ñ§â§Ñ§ä?§Ó, §ë§à ?§ß§Ô?§Ò§å§ð§ä§î §Õ?§ð §è§î§à§Ô§à §Ò?§Ý§Ü§Ñ §ä§Ñ §Ò§Ý§à§Ü§å§ð§ä§î §á§â§à§Ý?§æ§Ö§â§Ñ§è?§ð (§â?§ã§ä) §á§å§ç§Ý§Ú§ß§ß§Ú§ç §Ü§Ý?§ä§Ú§ß. ? §¥§Ý§ñ §Ó§Ú§Ù§ß§Ñ§é§Ö§ß§ß§ñ §Ö§æ§Ö§Ü§ä§Ú§Ó§ß§à§ã§ä? §Õ?? §ä§Ñ§Ü§à§Ô§à §á§â§Ö§á§Ñ§â§Ñ§ä§å §ß§Ö§à§Ò§ç?§Õ§ß§à §á§â§à§Ó§Ö§ã§ä§Ú §Õ§à§ã§Ý?§Õ§Ø§Ö§ß§ß§ñ §Ñ§Þ§á§Ý?§æ?§Ü§Ñ§è?? (§Ù§Ò?§Ý§î§ê§Ö§ß§ß§ñ §Ü?§Ý§î§Ü§à§ã§ä? §Ü§à§á?§Û) §Ô§Ö§ß§Ñ Her2/neu.
§¢§Ñ§Ù§à§Ó§Ú§Û §Ñ§Ý§Ô§à§â§Ú§ä§Þ §á§â§à§Ó§Ö§Õ§Ö§ß§ß§ñ §Õ§à§ã§Ý?§Õ§Ø§Ö§ß§ß§ñ * §°§è?§ß§Ü§Ñ §ã§ä§å§á§Ö§ß§ñ §Ö§Ü§ã§á§â§Ö§ã?? Her2/neu (§Þ§Ö§ä§à§Õ ?§¤§·) Her2/neu (0/+1) §ß§Ö§Ô§Ñ§ä§Ú§Ó§ß§Ú§Û Her2/neu (3+) §á§à§Ù§Ú§ä§Ú§Ó§ß§Ú§Û Her2/neu (2+) §ã§å§Þ§ß?§Ó§ß§Ú§Û §¡§Þ§á§Ý?§æ?§Ü§Ñ§è?§ñ §Ô§Ö§ß§Ñ HER2/neu (FISH) *ASCO¨CCAP HER2 Test Guideline Recommendations (2018)
§®§Ñ§ä§Ö§â?§Ñ§Ý: §Ô?§ã§ä§à§Ý§à§Ô?§é§ß§Ú§Û (FFPE tissue), §á§Ñ§â§Ñ§æ?§ß§à§Ó? §Ù§â?§Ù§Ú §±?§Õ§â§Ñ§ç§å§ß§à§Ü §Þ?§ß?§Þ§å§Þ 20 §Ü§Ý?§ä§Ú§ß ?§ß§Ó§Ñ§Ù§Ú§Ó§ß§à§Ô§à §Ü§à§Þ§á§à§ß§Ö§ß§ä§å §á§å§ç§Ý§Ú§ß§Ú §±§â§à§Ò§Ñ: Dual color probe §¥§à§ã§Ý?§Õ§Ø§Ö§ß§ß§ñ §Ñ§Þ§á§Ý?§æ?§Ü§Ñ§è?? §Ô§Ö§ß§Ñ HER2/neu (FISH) §¢§Ñ§Ù§à§Ó§Ú§Û §Ñ§Ý§Ô§à§â§Ú§ä§Þ §á§â§à§Ó§Ö§Õ§Ö§ß§ß§ñ §Õ§à§ã§Ý?§Õ§Ø§Ö§ß§ß§ñ * HER2/CEN17 >2, HER2 >6 §á§à§Ù§Ú§ä§Ú§Ó§ß§Ú§Û HER2/CEP17 <2, HER2 = 4-6 §á§à§Ô§â§Ñ§ß§Ú§é§ß§Ú§Û HER2/CEN17 <2, HER2 <4 §ß§Ö§Ô§Ñ§ä§Ú§Ó§ß§Ú§Û *ASCO¨CCAP HER2 Test Guideline Recommendations (2018)
¡ñ §¤§Ö§ß ALK (Anaplastic lymphoma kinase) §â§à§Ù§ä§Ñ§ê§à§Ó§Ñ§ß§Ú§Û §ß§Ñ §Ü§à§â§à§ä§Ü§à§Þ§å §á§Ý§Ö§é? 2 §ç§â§à§Þ§à§ã§à§Þ§Ú ¡ñ §¤§Ö§ß ALK §Ü§à§Õ§å? §ä§â§Ñ§ß§ã§Þ§Ö§Þ§Ò§â§Ñ§ß§ß§Ú§Û §Ò?§Ý§à§Ü §Ù §ä§Ú§â§à§Ù§Ú§ß§Ü?§ß§Ñ§Ù§ß§à§ð §Ñ§Ü§ä§Ú§Ó§ß?§ã§ä§ð, §ñ§Ü§Ú§Û ? §Þ?§ê§Ö§ß§ß§ð §Õ§Ý§ñ §Õ?? §ä§Ñ§â§Ô§Ö§ä§ß§Ú§ç §á§â§Ö§á§Ñ§â§Ñ§ä?§Ó, ?§ß§Ô?§Ò?§ä§à§â?§Ó ALK. ¡ñ §´§â§Ñ§ß§ã§Ý§à§Ü§Ñ§è?§ñ §Ô§Ö§ß§å ALK §ã§á§à§ã§ä§Ö§â?§Ô§Ñ?§ä§î§ã§ñ §å 5-7% §á§Ñ§è??§ß§ä?§Ó §Ù §ß§Ö§Õ§â?§Ò§ß§à§Ü§Ý?§ä§Ú§ß§ß§Ú§Þ §â§Ñ§Ü§à§Þ §Ý§Ö§Ô§Ö§ß§î (§¯§¥§²§­) §£§Ú§Ù§ß§Ñ§é§Ö§ß§ß§ñ §é§å§ä§Ý§Ú§Ó§à§ã§ä? §Õ§à §ä§Ñ§â§Ô§Ö§ä§ß§Ú§ç §á§â§Ö§á§Ñ§â§Ñ§ä?§Ó §£§Ú§ñ§Ó§Ý§Ö§ß§ß§ñ §ä§â§Ñ§ß§ã§Ý§à§Ü§Ñ§è?? §Ô§Ö§ß§Ñ ALK (§¡§Õ§Ö§ß§à§Ü§Ñ§â§è§Ú§ß§à§Þ§Ñ §Ý§Ö§Ô§Ö§ß§î)
§®§Ñ§ä§Ö§â?§Ñ§Ý: §Ô?§ã§ä§à§Ý§à§Ô?§é§ß§Ú§Û §Þ§Ñ§ä§Ö§â?§Ñ§Ý (FFPE tissue), §è§Ú§ä§à§Ý§à§Ô?§é§ß§Ú§Û §Þ§Ñ§ä§Ö§â?§Ñ§Ý §±?§Õ§â§Ñ§ç§å§ß§à§Ü §Þ?§ß?§Þ§å§Þ 50 §á§å§ç§Ý§Ú§ß§ß§Ú§ç §Ü§Ý?§ä§Ú§ß §±§â§à§Ò§Ñ: Dual color probe (break apart) §²§Ö§Ü§à§Þ§Ö§ß§Õ§Ñ§è?? §ë§à§Õ§à ?§ß§ä§Ö§â§á§â§Ö§ä§Ñ§è?? * *The IASLC Atlas of ALK and ROS1 Testing in Lung Cancer
§£§Ú§Ù§ß§Ñ§é§Ö§ß§ß§ñ §Ô§â§å§á §â§Ú§Ù§Ú§Ü§å §®§Ñ§ä§Ö§â?§Ñ§Ý: §Ü§â§à§Ó §Ñ§Ò§à §Ñ§ã§á?§â§Ñ§ä §Ü?§ã§ä§Ü§à§Ó§à§Ô§à §Þ§à§Ù§Ü§å §ß§Ñ EDTA §±?§Õ§â§Ñ§ç§å§ß§à§Ü §Þ?§ß?§Þ§å§Þ 100 §Ü§Ý?§ä§Ú§ß Cut off ¨C 10 % §±§â§à§Ò§Ñ: Dual color probe §£§Ú§ñ§Ó§Ý§Ö§ß§ß§ñ §Õ§Ö§Ý§Ö§è?? 17p13.1 (TP53) ? §¤§Ö§ß TP53 §â§à§Ù§ä§Ñ§ê§à§Ó§Ñ§ß§Ú§Û §ß§Ñ §Ü§à§â§à§ä§Ü§à§Þ§å §á§Ý§Ö§é? 17 §ç§â§à§Þ§à§ã§à§Þ§Ú (17p13.1) ? §¤§Ö§ß TP53 §Ü§à§Õ§å? §Ò?§Ý§à§Ü 53 §Ü§¥§Ñ (§â53), §ë§à ? §ä§â§Ñ§ß§ã§Ü§â§Ú§á§è?§Û§ß§Ú§Þ §æ§Ñ§Ü§ä§à§â§à§Þ §ä§Ñ §Ò§Ö§â§Ö §å§é§Ñ§ã§ä§î §Ó §â§Ö§Ô§å§Ý§ñ§è?? §Ü§Ý?§ä§Ú§ß§ß§à? §á§â§à§Ý?§æ§Ö§â§Ñ§è??, §Õ§Ú§æ§Ö§â§Ö§ß§è?§Ñ§è?? §ä§Ñ §Ñ§á§à§á§ä§à§Ù§å. ? §ã§å§á§â§Ö§ã§à§â§à§Þ §á§å§ç§Ý§Ú§ß§ß§à§Ô§à §â§à§ã§ä§å. ? §¥§Ö§Ý§Ö§è?§ñ 17p13.1 (TP53) §Ù§å§ã§ä§â?§é§Ñ?§ä§î§ã§ñ §á§â§Ú §Ò§Ñ§Ô§Ñ§ä§î§à§ç §à§ß§Ü§à§Ô§Ö§Þ§Ñ§ä§à§Ý§à§Ô?§é§ß§Ú§ç §Ù§Ñ§ç§Ó§à§â§ð§Ó§Ñ§ß§ß§ñ§ç (§·§­§­, §®§¥§³, §Þ§ß§à§Ø§Ú§ß§ß§Ñ §Þ??§Ý§à§Þ§Ñ §ä§Ñ ?§ß.) ? §Ù§Ñ§Ù§Ó§Ú§é§Ñ§Û §Ñ§ã§à§è?§ð?§ä§î§ã§ñ §Ù §ß§Ö§ã§á§â§Ú§ñ§ä§Ý§Ú§Ó§Ú§Þ §á§â§à§Ô§ß§à§Ù§à§Þ (§Ô§â§å§á§Ñ §Ó§Ú§ã§à§Ü§à§Ô§à §â§Ú§Ù§Ú§Ü§å)
§°§ä§Ø§Ö, ¡ñ FISH ? §Ó§Ñ§Ø§Ý§Ú§Ó§Ú§Þ §Þ§Ö§ä§à§Õ§à§Þ §Þ§à§Ý§Ö§Ü§å§Ý§ñ§â§ß§à-§è§Ú§ä§à§Ô§Ö§ß§Ö§ä§Ú§é§ß§Ú§ç §Õ§à§ã§Ý?§Õ§Ø§Ö§ß§î, §ñ§Ü§Ú§Û §Õ§à§Ù§Ó§à§Ý§ñ? §Ó§Ú§ñ§Ó§Ú§ä§Ú §â?§Ù§ß? §Ô§Ö§ß§Ö§ä§Ú§é§ß? §á§à§â§å§ê§Ö§ß§ß§ñ (§ä§â§Ñ§ß§ã§Ý§à§Ü§Ñ§è??, §Õ§Ö§Ý§Ö§è??, §Ñ§Þ§á§Ý?§æ?§Ü§Ñ§è?? §ä§à§ë§à) ¡ñ ? §Ó§Ñ§Ø§Ý§Ú§Ó§Ú§Þ §Õ§à§á§à§Þ?§Ø§ß§Ú§Þ ?§ß§ã§ä§â§å§Þ§Ö§ß§ä§à§Þ §Ó 1. §Õ?§Ñ§Ô§ß§à§ã§ä§Ú§è? §à§ß§Ü§à§Ý§à§Ô?§é§ß§Ú§ç §Ù§Ñ§ç§Ó§à§â§ð§Ó§Ñ§ß§î §ä§Ñ §á?§Õ§ä§Ó§Ö§â§Õ§Ø§Ö§ß? §Õ?§Ñ§Ô§ß§à§Ù§å 2. §á?§Õ§Ò§à§â? §ä§Ñ§Ü§ä§Ú§Ü§Ú §Ý?§Ü§å§Ó§Ñ§ß§ß§ñ 3. §á§â§à§Ô§ß§à§Ù? §á§Ö§â§Ö§Ò?§Ô§å §Ù§Ñ§ç§Ó§à§â§ð§Ó§Ñ§ß§ß§ñ
§¥§ñ§Ü§å§ð §Ù§Ñ §å§Ó§Ñ§Ô§å!

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§¶?§ä§î§à §¡.?. - §°§ã§ß§à§Ó§Ú §Þ§Ö§ä§à§Õ§å Fish

  • 1. §°§ã§ß§à§Ó§Ú §Þ§Ö§ä§à§Õ§å FISH, §Õ§à§ã§Ó?§Õ §Ó§Ú§Ü§à§â§Ú§ã§ä§Ñ§ß§ß§ñ §Ó §Ý§Ñ§Ò§à§â§Ñ§ä§à§â?? CSD §¢?§à§Ý§à§Ô §¶?§ä§î§à §¡§ß§Ñ§ã§ä§Ñ§ã?§ñ §¬§Ú?§Ó ¨C 2021
  • 2. FISH - Fluorescence in situ Hybridization §¤?§Ò§â§Ú§Õ§Ú§Ù§Ñ§è?§ñ in situ ¨C §Ò§Ñ§Ù§à§Ó§Ú§Û §Þ§Ö§ä§à§Õ §Þ§à§Ý§Ö§Ü§å§Ý§ñ§â§ß§à-§è§Ú§ä§à§Ô§Ö§ß§Ö§ä§Ú§é§ß§Ú§ç §Õ§à§ã§Ý?§Õ§Ø§Ö§ß§î. ¡ñ §Õ§à§Ù§Ó§à§Ý§ñ? §Ó§Ú§ñ§Ó§Ú§ä§Ú §ã§á§Ö§è§Ú§æ?§é§ß§å §á§à§ã§Ý?§Õ§à§Ó§ß?§ã§ä§î §ß§å§Ü§Ý§Ö?§ß§à§Ó§à? §Ü§Ú§ã§Ý§à§ä§Ú (§¥§¯§¬ §é§Ú §²§¯§¬) §Ó §Ü§Ý?§ä§Ú§ß?. ¡ñ §±§â§Ú§ß§è§Ú§á §Þ§Ö§ä§à§Õ§å §á§à§Ý§ñ§Ô§Ñ? §å §Ü§à§Þ§á§Ý§Ö§Þ§Ö§ß§ä§Ñ§â§ß§à§Þ§å §Ù§Ó¡¯§ñ§Ù§å§Ó§Ñ§ß§ß? §ß§å§Ü§Ý§Ö§à§ä§Ú§Õ§ß§à§Ô§à §Ù§à§ß§Õ§å (§á§â§à§Ò§Ú) ?§Ù §á§à§ã§Ý?§Õ§à§Ó§ß?§ã§ä§ð §¥§¯§¬ §é§Ú §²§¯§¬ §Õ§à§ã§Ý?§Õ§Ø§å§Ó§Ñ§ß§à§Ô§à §Ù§â§Ñ§Ù§Ü§Ñ. §¤?§Ò§â§Ú§Õ§Ú§Ù§Ñ§è?§ñ in situ §¶§à§â§Þ§å§Ó§Ñ§ß§ß§ñ §Ô?§Ò§â§Ú§Õ?§Ó (§¥§¯§¬-§¥§¯§¬, §¥§¯§¬-§²§¯§¬, §²§¯§¬-§²§¯§¬) §£ §å§Þ§à§Ó§Ñ§ç §æ?§Ü§ã§à§Ó§Ñ§ß§à§Ô§à §ã§å§Ò§ã§ä§â§Ñ§ä§å, §Ó §Ü§Ý?§ä§Ú§ß? §¥§¯§¬ §Ù§â§Ñ§Ù§Ü§Ñ §©§à§ß§Õ
  • 3. §¬§Ý§Ñ§ã§Ú§æ?§Ü§Ñ§è?§ñ §Ù§à§ß§Õ?§Ó ¡ñ §©§à§ß§Õ (§á§â§à§Ò§Ñ) ¨C §è§Ö §Ü§à§â§à§ä§Ü§Ñ §ß§å§Ü§Ý§Ö§à§ä§Ú§Õ§ß§Ñ §á§à§ã§Ý?§Õ§à§Ó§ß?§ã§ä§î (§à§á§ä§Ú§Þ§Ñ§Ý§î§ß§à 50-300 §á§Ñ§â §à§ã§ß§à§Ó), §Ü§à§Þ§á§Ý§Ö§Þ§Ö§ß§ä§Ñ§â§ß§Ñ §Õ§à§ã§Ý?§Õ§Ø§å§Ó§Ñ§ß?§Û §á§à§ã§Ý?§Õ§à§Ó§ß§à§ã§ä? §ß§å§Ü§Ý§Ö?§ß§à§Ó§à? §Ü§Ú§ã§Ý§à§ä§Ú. §©§Ñ §Þ?§ã§è§Ö§Þ §á§â§Ú?§Õ§ß§Ñ§ß§ß§ñ: a) §Ý§à§Ü§å§ã§ã§á§Ö§è§Ú§æ?§é§ß? (LSI - Locus Specific Identifier) b) §è§Ö§ß§ä§â§à§Þ§Ö§â§ß? (CEP) c) §ä§Ö§Ý§à§Þ§Ö§â§ß? d) §Õ§à §Ó§ã??? §ç§â§à§Þ§à§ã§à§Þ§Ú (WCP - Whole chromosome painting) §©§Ñ §Ò§å§Õ§à§Ó§à§ð: a) §Õ§Ó§à§Ý§Ñ§ß§è§ð§Ô§à§Ó? §¥§¯§¬-§Ù§à§ß§Õ§Ú, b) §à§Õ§ß§à§Ý§Ñ§ß§è§ð§Ô§à§Ó? §Ñ§ß§ä§Ú§ã§Ö§ß§ã-§²§¯§¬-§Ù§à§ß§Õ§Ú (§â§Ú§Ò§à§á§â§à§Ò§Ú), c) §à§Õ§ß§à§Ý§Ñ§ß§è§ð§Ô§à§Ó? §¥§¯§¬-§Ù§à§ß§Õ§Ú, d) §ã§Ú§ß§ä§Ö§ä§Ú§é§ß? §à§Ý?§Ô§à§Õ§Ö§Ù§à§Ü§ã§Ú§ß§å§Ü§Ý§Ö§à§ä§Ú§Õ§ß? §Ù§à§ß§Õ§Ú §ä§Ñ §à§Ý?§Ô§à§â§Ú§Ò§à§Ù§à§ß§Õ§Ú
  • 4. §©§Ñ §ã§á§à§ã§à§Ò§à§Þ §Þ?§é§Ö§ß§ß§ñ: §©§à§ß§Õ§Ú §© §â§Ñ§Õ?§à§Ñ§Ü§ä§Ú§Ó§ß§à§ð §Þ?§ä§Ü§à§ð §© §ß§Ö§â§Ñ§Õ?§à§Ñ§Ü§ä§Ú§Ó§ß§à§ð §Þ?§ä§Ü§à§ð P32, S35, H3 §¢?§à§ä§Ú§ß §¥§Ú§Ô§à§Ü§ã§Ú§Ô§Ö§ß?§ß (DIG) §¶§Ý§å§à§â§à§ç§â§à§Þ §Þ?§ä§Ü§Ú fluorescein rhodamine coumarin Texas Red SpectrumOrange SpectrumGreen
  • 5. §£§Ú§Õ§Ú §Ô?§Ò§â§Ú§Õ§Ú§Ù§Ñ§è?? in situ ISH §²§Ñ§Õ?§à§Ñ§Ü§ä§Ú§Ó§ß§Ú§Û (§Ñ§Ó§ä§à§â§Ñ§Õ?§à§Ô§â§Ñ§æ?§ñ) BRISH Bright field in situ hybridization (§ã§Ó?§ä§Ý§à§Ó§Ñ §Þ?§Ü§â§à§ã§Ü§à§á?§ñ) §³ISH Chromogenic in situ hybridization SISH Silver in situ hybridization ?§ß§ê? FISH Fluorescent in situ hybridization (§æ§Ý§å§à§â§Ö§ã§è§Ö§ß§ä§ß§Ñ §Þ?§Ü§â§à§ã§Ü§à§á?§ñ) Rainbow-FISH Raman-FISH ReD-FISH Reverse-FISH RING-FISH RNA-FISH RxFISH Split-Signal FISH T-FISH 3-D FISH Zoo-FISH ACM-FISH armFISH CARD-FISH catFISH CB-FISH CO-FISH COBRA-FISH COD-FISH COMBO-FISH Comet-FISH Cryo-FISH D-FISH DBD-FISH e-FISH Fiber-FISH Flow-FISH Fusion-Signal FISH Halo-FISH Harlequin-FISH Immuno-FISH LNA-FISH M-FISH Multilocus or ML-FISH PCC-FISH PNA-FISH Q-FISH QD-FISH
  • 6. §±§â§Ú§ß§è§Ú§á §ä§Ñ §à§ã§ß§à§Ó§ß? §Ö§ä§Ñ§á§Ú §Þ§Ö§ä§à§Õ§å FISH/CISH (§ß§Ñ §á§â§Ú§Ü§Ý§Ñ§Õ? §Ô?§ã§ä§à§Ý§à§Ô?§é§ß§à§Ô§à §Þ§Ñ§ä§Ö§â?§Ñ§Ý§å) §±§Ñ§â§Ñ§æ?§ß§à§Ó§Ú§Û §Ò§Ý§à§Ü §Ù §æ?§Ü§ã§à§Ó§Ñ§ß§à§ð §å §æ§à§â§Þ§Ñ§Ý?§ß? §ä§Ü§Ñ§ß§Ú§ß§à§ð §£§Ú§Ô§à§ä§à§Ó§Ý§Ö§ß§ß§ñ §Ù§â?§Ù?§Ó §ä§à§Ó§ë§Ú§ß§à§ð 2-3 ¦Ìm §¥§Ö§á§Ñ§â§Ñ§æ?§ß?§Ù§Ñ§è?§ñ §å §Ü§ã§Ú§Ý§à§Ý? §¥§Ö§Ô?§Õ§â§Ñ§ä§Ñ§è?§ñ §å §ã§Ö§â?? §ã§á§Ú§â§ä?§Ó §Ó §á§à§â§ñ§Õ§Ü§å §Ù§Þ§Ö§ß§ê§Ö§ß§ß§ñ ?§ç §Ü§à§ß§è§Ö§ß§ä§â§Ñ§è?? §¶§Ö§â§Þ§Ö§ß§ä§Ñ§ä§Ú§Ó§ß§Ú§Û §á§â§à§ä§Ö§à§Ý?§Ù (§ß§Ñ§Û§é§Ñ§ã§ä?§ê§Ö §Ù §Ó§Ú§Ü§à§â§Ú§ã§ä§Ñ§ß§ß§ñ§Þ §á§Ö§á§ã§Ú§ß§å) §¥§à§Õ§Ñ§Ó§Ñ§ß§ß§ñ §Þ?§é§Ö§ß§à? §á§â§à§Ò§Ú §¥§Ö§ß§Ñ§ä§å§â§Ñ§è?§ñ (82 ¡ã§³, 5 §ç§Ó) §¤?§Ò§â§Ú§Õ§Ú§Ù§Ñ§è?§ñ 37 ¡ã§³ (18 §Ô§à§Õ) §±§à§ã§ä§Ô?§Ò§â§Ú§Õ§Ú§Ù§Ñ§è?§Û§ß§Ñ §Ó?§Õ§Þ§Ú§Ó§Ü§Ñ §¥§Ö§ä§Ö§Ü§è?§ñ §±§â§ñ§Þ§Ñ (§á§â§à§Ò§Ñ, §Þ?§é§Ö§ß§Ñ §æ§Ý§å§à§â§à§ç§â§à§Þ§à§Þ, FISH §Þ§Ö§ä§à§Õ) §¯§Ö§á§â§ñ§Þ§Ñ (§á§â§à§Ò§Ñ, §Þ?§é§Ö§ß§Ñ §Ò?§à§ä§Ú§ß§à§Þ §Ñ§Ò§à §Õ§Ú§Ô§à§Ü§ã§Ú§Ô§Ö§ß?§ß§à§Þ, §³ISH §Þ§Ö§ä§à§Õ) §¬§à§ß§ä§â§æ§Ñ§â§Ò§å§Ó§Ñ§ß§ß§ñ §Ù 4¡¯,6-§Õ?§Ñ§Þ?§ß§à-2- §æ§Ö§ß?§Ý?§ß§Õ§à§Ý§à§Þ (DAPI)
  • 7. §±§â§Ú§ß§è§Ú§á §Þ§Ö§ä§à§Õ§å CISH HRP ¨C Horseradish peroxidase (§á§Ö§â§à§Ü§ã§Ú§Õ§Ñ§Ù§Ñ §ç§â§à§ß§å) DAB ¨C Diaminobenzidine ¨C §ã§å§Ò§ã§ä§â§Ñ§ä §Õ§Ý§ñ HRP EBV (CISH)
  • 8. ¡ñ §±§â§à§Ó§à§Õ§Ú§ä§î§ã§ñ §Ù§Ñ §Õ§à§á§à§Þ§à§Ô§à§ð §æ§Ý§å§à§â§Ö§ã§è§Ö§ß§ä§ß§à§Ô§à §Þ?§Ü§â§à§ã§Ü§à§á§Ñ. ¡ñ §¡§ß§Ñ§Ý?§Ù ? §á?§Õ§â§Ñ§ç§å§ß§à§Ü §Ü§Ý?§ä§Ú§ß §Ó?§Õ§Ò§å§Ó§Ñ?§ä§ã§î§ñ §á§â§Ú §Ó§Ö§Ý§Ú§Ü§à§Þ§å §Ù§Ò?§Ý§î§ê§Ö§ß§ß? (10§ç100) §Ù §Ó§Ú§Ü§à§â§Ú§ã§ä§Ñ§ß§ß§ñ§Þ ?§Þ§Ö§â§ã?§Û§ß§à? §à§Ý?? §Õ§Ý§ñ §æ§Ý§å§à§â§Ö§ã§è§Ö§ß§ä§ß§à§Ô§à §Þ?§Ü§â§à§ã§Ü§à§á§Ñ (Type FF) ?§ß§ä§Ö§â§á§â§Ö§ä§Ñ§è?§ñ §â§Ö§Ù§å§Ý§î§ä§Ñ§ä?§Ó FISH DAPI Green/orange §æ?§Ý§î§ä§â
  • 9. a) Single color probe b) Dual color probe c) Multi color probe d) Dual color Break apart probe e) Dual color Fusion probe Dual Color Probe Multi color probe Dual color Fusion probe Dual Color Break Apart Probe §°§á§Ú§ã (§Ü§Ñ§â§ä§Ñ) §á§â§à§Ò§Ú
  • 10. §±§â§Ñ§Ü§ä§Ú§é§ß§Ö §Ù§Ñ§ã§ä§à§ã§å§Ó§Ñ§ß§ß§ñ FISH (CISH) §Ó §Þ§Ö§Õ§Ú§è§Ú§ß? ¡ñ §£§Ú§ñ§Ó§Ý§Ö§ß§ß§ñ §Ó?§â§å§ã§ß§Ú§ç §Ô§Ö§ß§à§Þ?§Ó (EBV, HPV) ¡ñ §£§Ú§ñ§Ó§Ý§Ö§ß§ß§ñ §Ô§Ö§ß§Ö§ä§Ú§é§ß§Ú§ç §á§à§â§å§ê§Ö§ß§î §ß§Ñ §Þ§Ö§ä§Ñ§æ§Ñ§Ù§ß§Ú§ç §ç§â§à§Þ§à§ã§à§Þ§Ñ§ç §Ñ§Ò§à §Ó ?§ß§ä§Ö§â§æ§Ñ§Ù§ß§Ú§ç §ñ§Õ§â§Ñ§ç §Ü§Ý?§ä§Ú§ß (§ç§â§à§Þ§à§ã§à§Þ§ß? §Ñ§Ò§Ö§â§Ñ§è??, §ä§â§Ñ§ß§ã§Ý§à§Ü§Ñ§è??, §Õ§Ö§Ý§Ö§è??, §Ñ§Þ§á§Ý?§æ?§Ü§Ñ§è?? §ä§à§ë§à) ¡ñ §¥§Ú§æ§Ö§â§Ö§ß§è?§Û§ß§Ñ §Õ?§Ñ§Ô§ß§à§ã§ä§Ú§Ü§Ñ §Ó §à§ß§Ü§à§Ý§à§Ô?? ¡ñ §£§Ú§Ù§ß§Ñ§é§Ö§ß§ß§ñ §é§å§ä§Ý§Ú§Ó§à§ã§ä? §Õ§à §ä§Ñ§â§Ô§Ö§ä§ß§Ú§ç §á§â§Ö§á§Ñ§â§Ñ§ä?§Ó §Ó §à§ß§Ü§à§Ý§à§Ô?? ¡ñ §£§Ú§Ù§ß§Ñ§é§Ö§ß§ß§ñ §Ô§â§å§á §â§Ú§Ù§Ú§Ü§å §Ó §à§ß§Ü§à§Ý§à§Ô?? ¡ñ §±§â§Ö§ß§Ñ§ä§Ñ§Ý§î§ß§Ñ §Õ?§Ñ§Ô§ß§à§ã§ä§Ú§Ü§Ñ
  • 11. §±§â§Ú§Ü§Ý§Ñ§Õ§Ú §Ù§Ñ§ã§ä§à§ã§å§Ó§Ñ§ß§ß§ñ §Þ§Ö§ä§à§Õ§å FISH §Ó §Ý§Ñ§Ò§à§â§Ñ§ä§à§â?? CSD ? §£ 1960 §Ô Peter Nowell §ä§Ñ David Hungerford §à§á§Ú§ã§Ñ§Ý§Ú §ã§á§Ö§è§Ú§æ?§é§ß§å §ç§â§à§Þ§à§ã§à§Þ§å, §ñ§Ü§å §ß§Ñ§Ù§Ó§Ñ§Ý§Ú §æ?§Ý§Ñ§Õ§Ö§Ý§î§æ?§Û§ã§î§Ü§à§ð. ? §¶?§Ý§Ñ§Õ§Ö§Ý§î§æ?§Û§ã§î§Ü§Ñ §ç§â§à§Þ§à§ã§à§Þ§Ñ (Ph.) ¨C §è§Ö §â§Ö§Ù§å§Ý§î§ä§Ñ§ä §â§Ö§è§Ú§á§â§à§Ü§ß§à? §ä§â§Ñ§ß§ã§Ý§à§Ü§Ñ§è??* §Þ?§Ø §Õ?§Ý§ñ§ß§Ü§Ñ§Þ§Ú §Õ§à§Ó§Ô§à§Ô§à §á§Ý§Ö§é§Ñ 22 (§Õ?§Ý§ñ§ß§Ü§Ñ 22q11.23, §Ô§Ö§ß BCR) §ä§Ñ 9 (§Õ?§Ý§ñ§ß§Ü§Ñ 9q34.12, §Ô§Ö§ß ABL1) §ç§â§à§Þ§à§ã§à§Þ§Ú. ? §µ 90% §á§Ñ§è??§ß§ä?§Ó §Ù §ç§â§à§ß?§é§ß§Ú§Þ §Þ??§Ý§à§Ý§Ö§Û§Ü§à§Ù§à§Þ ? 25% §Ù §Ô§à§ã§ä§â§Ú§Þ §Ý?§Þ§æ§à§Ò§Ý§Ñ§ã§ä§ß§Ú§Þ §Ý§Ö§Û§Ü§à§Ù§à§Þ §ß§Ñ§ñ§Ó§ß§Ñ §ä§â§Ñ§ß§ã§Ý§à§Ü§Ñ§è?§ñ ? BCR/ABL t (9; 22) (q34.1; q11.2). *§²§Ö§è§Ú§á§â§à§Ü§ß§Ñ §ä§â§Ñ§ß§ã§Ý§à§Ü§Ñ§è?§ñ - §à§Ò§Þ?§ß §Õ?§Ý§ñ§ß§Ü§Ñ§Þ§Ú §Þ?§Ø §Õ§Ó§à§Þ§Ñ §ß§Ö§Ô§à§Þ§à§Ý§à§Ô?§é§ß§Ú§Þ§Ú §ç§â§à§Þ§à§ã§à§Þ§Ñ§Þ§Ú §£§Ú§Ù§ß§Ñ§é§Ö§ß§ß§ñ ?§¶?§Ý§Ñ§Õ§Ö§Ý§î§æ?§Û§ã§î§Ü§à? §ç§â§à§Þ§à§ã§à§Þ§Ú? (BCR/ABL, t (9; 22) (q34.1; q11.2))
  • 12. BCR/ABL - (2R2G) BCR/ABL + (2F1R1G) BCR/ABL + (1F1R1G) §®§Ñ§ä§Ö§â?§Ñ§Ý: §Ü§â§à§Ó §Ñ§Ò§à §Ñ§ã§á?§â§Ñ§ä §Ü?§ã§ä§Ü§à§Ó§à§Ô§à §Þ§à§Ù§Ü§å §±?§Õ§â§Ñ§ç§å§ß§à§Ü §Þ?§ß?§Þ§å§Þ 100 §Ü§Ý?§ä§Ú§ß Cut off ¨C 2 % §±§â§à§Ò§Ñ: Dual color Fusion probe §£§Ú§Ù§ß§Ñ§é§Ö§ß§ß§ñ §æ?§Ý§Ñ§Õ§Ö§Ý§î§æ?§Û§ã§î§Ü§à? §ç§â§à§Þ§à§ã§à§Þ§Ú BCR/ABL
  • 13. §£§Ú§Ù§ß§Ñ§é§Ö§ß§ß§ñ §é§å§ä§Ý§Ú§Ó§à§ã§ä? §Õ§à §ä§Ñ§â§Ô§Ö§ä§ß§Ú§ç §á§â§Ö§á§Ñ§â§Ñ§ä?§Ó §¥§à§ã§Ý?§Õ§Ø§Ö§ß§ß§ñ §Ñ§Þ§á§Ý?§æ?§Ü§Ñ§è?? §Ô§Ö§ß§Ñ Her2/neu (§â§Ñ§Ü §Þ§à§Ý§à§é§ß§à? §Ù§Ñ§Ý§à§Ù§Ú, §â§Ñ§Ü §ê§Ý§å§ß§Ü§Ñ, §Ü§à§Ý§à§â§Ö§Ü§ä§Ñ§Ý§î§ß§Ú§Û §â§Ñ§Ü) ? §¤§Ö§ß ERBB2 (Her2/neu) §â§à§Ù§ä§Ñ§ê§à§Ó§Ñ§ß§Ú§Û §ß§Ñ §Õ§à§Ó§Ô§à§Þ§å §á§Ý§Ö§é? 17 §ç§â§à§Þ§à§ã§à§Þ§Ú (17q12) ? §¤§Ö§ß ERBB2 §Ü§à§Õ§å? §Ò?§Ý§à§Ü 185-190 §Ü§¥§Ñ (Her2/neu), §ë§à ? §â§Ö§è§Ö§á§ä§à§â§à§Þ §Ù §ä§Ú§â§à§Ù§Ú§ß§Ü?§ß§Ñ§Ù§ß§à§ð §Ñ§Ü§ä§Ú§Ó§ß?§ã§ä§ð §ä§Ñ §Ò§Ö§â§Ö §å§é§Ñ§ã§ä§î §Ó §â§Ö§Ô§å§Ý§ñ§è?? §Ü§Ý?§ä§Ú§ß§ß§à§Ô§à §â§à§ã§ä§å. ? §¢?§Ý§à§Ü Her2/neu ? §Þ?§ê§Ö§ß§ß§ð §Õ§Ý§ñ §ä§Ñ§â§Ô§Ö§ä§ß§Ú§ç §á§â§Ö§á§Ñ§â§Ñ§ä?§Ó, §ë§à ?§ß§Ô?§Ò§å§ð§ä§î §Õ?§ð §è§î§à§Ô§à §Ò?§Ý§Ü§Ñ §ä§Ñ §Ò§Ý§à§Ü§å§ð§ä§î §á§â§à§Ý?§æ§Ö§â§Ñ§è?§ð (§â?§ã§ä) §á§å§ç§Ý§Ú§ß§ß§Ú§ç §Ü§Ý?§ä§Ú§ß. ? §¥§Ý§ñ §Ó§Ú§Ù§ß§Ñ§é§Ö§ß§ß§ñ §Ö§æ§Ö§Ü§ä§Ú§Ó§ß§à§ã§ä? §Õ?? §ä§Ñ§Ü§à§Ô§à §á§â§Ö§á§Ñ§â§Ñ§ä§å §ß§Ö§à§Ò§ç?§Õ§ß§à §á§â§à§Ó§Ö§ã§ä§Ú §Õ§à§ã§Ý?§Õ§Ø§Ö§ß§ß§ñ §Ñ§Þ§á§Ý?§æ?§Ü§Ñ§è?? (§Ù§Ò?§Ý§î§ê§Ö§ß§ß§ñ §Ü?§Ý§î§Ü§à§ã§ä? §Ü§à§á?§Û) §Ô§Ö§ß§Ñ Her2/neu.
  • 14. §¢§Ñ§Ù§à§Ó§Ú§Û §Ñ§Ý§Ô§à§â§Ú§ä§Þ §á§â§à§Ó§Ö§Õ§Ö§ß§ß§ñ §Õ§à§ã§Ý?§Õ§Ø§Ö§ß§ß§ñ * §°§è?§ß§Ü§Ñ §ã§ä§å§á§Ö§ß§ñ §Ö§Ü§ã§á§â§Ö§ã?? Her2/neu (§Þ§Ö§ä§à§Õ ?§¤§·) Her2/neu (0/+1) §ß§Ö§Ô§Ñ§ä§Ú§Ó§ß§Ú§Û Her2/neu (3+) §á§à§Ù§Ú§ä§Ú§Ó§ß§Ú§Û Her2/neu (2+) §ã§å§Þ§ß?§Ó§ß§Ú§Û §¡§Þ§á§Ý?§æ?§Ü§Ñ§è?§ñ §Ô§Ö§ß§Ñ HER2/neu (FISH) *ASCO¨CCAP HER2 Test Guideline Recommendations (2018)
  • 15. §®§Ñ§ä§Ö§â?§Ñ§Ý: §Ô?§ã§ä§à§Ý§à§Ô?§é§ß§Ú§Û (FFPE tissue), §á§Ñ§â§Ñ§æ?§ß§à§Ó? §Ù§â?§Ù§Ú §±?§Õ§â§Ñ§ç§å§ß§à§Ü §Þ?§ß?§Þ§å§Þ 20 §Ü§Ý?§ä§Ú§ß ?§ß§Ó§Ñ§Ù§Ú§Ó§ß§à§Ô§à §Ü§à§Þ§á§à§ß§Ö§ß§ä§å §á§å§ç§Ý§Ú§ß§Ú §±§â§à§Ò§Ñ: Dual color probe §¥§à§ã§Ý?§Õ§Ø§Ö§ß§ß§ñ §Ñ§Þ§á§Ý?§æ?§Ü§Ñ§è?? §Ô§Ö§ß§Ñ HER2/neu (FISH) §¢§Ñ§Ù§à§Ó§Ú§Û §Ñ§Ý§Ô§à§â§Ú§ä§Þ §á§â§à§Ó§Ö§Õ§Ö§ß§ß§ñ §Õ§à§ã§Ý?§Õ§Ø§Ö§ß§ß§ñ * HER2/CEN17 >2, HER2 >6 §á§à§Ù§Ú§ä§Ú§Ó§ß§Ú§Û HER2/CEP17 <2, HER2 = 4-6 §á§à§Ô§â§Ñ§ß§Ú§é§ß§Ú§Û HER2/CEN17 <2, HER2 <4 §ß§Ö§Ô§Ñ§ä§Ú§Ó§ß§Ú§Û *ASCO¨CCAP HER2 Test Guideline Recommendations (2018)
  • 16. ¡ñ §¤§Ö§ß ALK (Anaplastic lymphoma kinase) §â§à§Ù§ä§Ñ§ê§à§Ó§Ñ§ß§Ú§Û §ß§Ñ §Ü§à§â§à§ä§Ü§à§Þ§å §á§Ý§Ö§é? 2 §ç§â§à§Þ§à§ã§à§Þ§Ú ¡ñ §¤§Ö§ß ALK §Ü§à§Õ§å? §ä§â§Ñ§ß§ã§Þ§Ö§Þ§Ò§â§Ñ§ß§ß§Ú§Û §Ò?§Ý§à§Ü §Ù §ä§Ú§â§à§Ù§Ú§ß§Ü?§ß§Ñ§Ù§ß§à§ð §Ñ§Ü§ä§Ú§Ó§ß?§ã§ä§ð, §ñ§Ü§Ú§Û ? §Þ?§ê§Ö§ß§ß§ð §Õ§Ý§ñ §Õ?? §ä§Ñ§â§Ô§Ö§ä§ß§Ú§ç §á§â§Ö§á§Ñ§â§Ñ§ä?§Ó, ?§ß§Ô?§Ò?§ä§à§â?§Ó ALK. ¡ñ §´§â§Ñ§ß§ã§Ý§à§Ü§Ñ§è?§ñ §Ô§Ö§ß§å ALK §ã§á§à§ã§ä§Ö§â?§Ô§Ñ?§ä§î§ã§ñ §å 5-7% §á§Ñ§è??§ß§ä?§Ó §Ù §ß§Ö§Õ§â?§Ò§ß§à§Ü§Ý?§ä§Ú§ß§ß§Ú§Þ §â§Ñ§Ü§à§Þ §Ý§Ö§Ô§Ö§ß§î (§¯§¥§²§­) §£§Ú§Ù§ß§Ñ§é§Ö§ß§ß§ñ §é§å§ä§Ý§Ú§Ó§à§ã§ä? §Õ§à §ä§Ñ§â§Ô§Ö§ä§ß§Ú§ç §á§â§Ö§á§Ñ§â§Ñ§ä?§Ó §£§Ú§ñ§Ó§Ý§Ö§ß§ß§ñ §ä§â§Ñ§ß§ã§Ý§à§Ü§Ñ§è?? §Ô§Ö§ß§Ñ ALK (§¡§Õ§Ö§ß§à§Ü§Ñ§â§è§Ú§ß§à§Þ§Ñ §Ý§Ö§Ô§Ö§ß§î)
  • 17. §®§Ñ§ä§Ö§â?§Ñ§Ý: §Ô?§ã§ä§à§Ý§à§Ô?§é§ß§Ú§Û §Þ§Ñ§ä§Ö§â?§Ñ§Ý (FFPE tissue), §è§Ú§ä§à§Ý§à§Ô?§é§ß§Ú§Û §Þ§Ñ§ä§Ö§â?§Ñ§Ý §±?§Õ§â§Ñ§ç§å§ß§à§Ü §Þ?§ß?§Þ§å§Þ 50 §á§å§ç§Ý§Ú§ß§ß§Ú§ç §Ü§Ý?§ä§Ú§ß §±§â§à§Ò§Ñ: Dual color probe (break apart) §²§Ö§Ü§à§Þ§Ö§ß§Õ§Ñ§è?? §ë§à§Õ§à ?§ß§ä§Ö§â§á§â§Ö§ä§Ñ§è?? * *The IASLC Atlas of ALK and ROS1 Testing in Lung Cancer
  • 18. §£§Ú§Ù§ß§Ñ§é§Ö§ß§ß§ñ §Ô§â§å§á §â§Ú§Ù§Ú§Ü§å §®§Ñ§ä§Ö§â?§Ñ§Ý: §Ü§â§à§Ó §Ñ§Ò§à §Ñ§ã§á?§â§Ñ§ä §Ü?§ã§ä§Ü§à§Ó§à§Ô§à §Þ§à§Ù§Ü§å §ß§Ñ EDTA §±?§Õ§â§Ñ§ç§å§ß§à§Ü §Þ?§ß?§Þ§å§Þ 100 §Ü§Ý?§ä§Ú§ß Cut off ¨C 10 % §±§â§à§Ò§Ñ: Dual color probe §£§Ú§ñ§Ó§Ý§Ö§ß§ß§ñ §Õ§Ö§Ý§Ö§è?? 17p13.1 (TP53) ? §¤§Ö§ß TP53 §â§à§Ù§ä§Ñ§ê§à§Ó§Ñ§ß§Ú§Û §ß§Ñ §Ü§à§â§à§ä§Ü§à§Þ§å §á§Ý§Ö§é? 17 §ç§â§à§Þ§à§ã§à§Þ§Ú (17p13.1) ? §¤§Ö§ß TP53 §Ü§à§Õ§å? §Ò?§Ý§à§Ü 53 §Ü§¥§Ñ (§â53), §ë§à ? §ä§â§Ñ§ß§ã§Ü§â§Ú§á§è?§Û§ß§Ú§Þ §æ§Ñ§Ü§ä§à§â§à§Þ §ä§Ñ §Ò§Ö§â§Ö §å§é§Ñ§ã§ä§î §Ó §â§Ö§Ô§å§Ý§ñ§è?? §Ü§Ý?§ä§Ú§ß§ß§à? §á§â§à§Ý?§æ§Ö§â§Ñ§è??, §Õ§Ú§æ§Ö§â§Ö§ß§è?§Ñ§è?? §ä§Ñ §Ñ§á§à§á§ä§à§Ù§å. ? §ã§å§á§â§Ö§ã§à§â§à§Þ §á§å§ç§Ý§Ú§ß§ß§à§Ô§à §â§à§ã§ä§å. ? §¥§Ö§Ý§Ö§è?§ñ 17p13.1 (TP53) §Ù§å§ã§ä§â?§é§Ñ?§ä§î§ã§ñ §á§â§Ú §Ò§Ñ§Ô§Ñ§ä§î§à§ç §à§ß§Ü§à§Ô§Ö§Þ§Ñ§ä§à§Ý§à§Ô?§é§ß§Ú§ç §Ù§Ñ§ç§Ó§à§â§ð§Ó§Ñ§ß§ß§ñ§ç (§·§­§­, §®§¥§³, §Þ§ß§à§Ø§Ú§ß§ß§Ñ §Þ??§Ý§à§Þ§Ñ §ä§Ñ ?§ß.) ? §Ù§Ñ§Ù§Ó§Ú§é§Ñ§Û §Ñ§ã§à§è?§ð?§ä§î§ã§ñ §Ù §ß§Ö§ã§á§â§Ú§ñ§ä§Ý§Ú§Ó§Ú§Þ §á§â§à§Ô§ß§à§Ù§à§Þ (§Ô§â§å§á§Ñ §Ó§Ú§ã§à§Ü§à§Ô§à §â§Ú§Ù§Ú§Ü§å)
  • 19. §°§ä§Ø§Ö, ¡ñ FISH ? §Ó§Ñ§Ø§Ý§Ú§Ó§Ú§Þ §Þ§Ö§ä§à§Õ§à§Þ §Þ§à§Ý§Ö§Ü§å§Ý§ñ§â§ß§à-§è§Ú§ä§à§Ô§Ö§ß§Ö§ä§Ú§é§ß§Ú§ç §Õ§à§ã§Ý?§Õ§Ø§Ö§ß§î, §ñ§Ü§Ú§Û §Õ§à§Ù§Ó§à§Ý§ñ? §Ó§Ú§ñ§Ó§Ú§ä§Ú §â?§Ù§ß? §Ô§Ö§ß§Ö§ä§Ú§é§ß? §á§à§â§å§ê§Ö§ß§ß§ñ (§ä§â§Ñ§ß§ã§Ý§à§Ü§Ñ§è??, §Õ§Ö§Ý§Ö§è??, §Ñ§Þ§á§Ý?§æ?§Ü§Ñ§è?? §ä§à§ë§à) ¡ñ ? §Ó§Ñ§Ø§Ý§Ú§Ó§Ú§Þ §Õ§à§á§à§Þ?§Ø§ß§Ú§Þ ?§ß§ã§ä§â§å§Þ§Ö§ß§ä§à§Þ §Ó 1. §Õ?§Ñ§Ô§ß§à§ã§ä§Ú§è? §à§ß§Ü§à§Ý§à§Ô?§é§ß§Ú§ç §Ù§Ñ§ç§Ó§à§â§ð§Ó§Ñ§ß§î §ä§Ñ §á?§Õ§ä§Ó§Ö§â§Õ§Ø§Ö§ß? §Õ?§Ñ§Ô§ß§à§Ù§å 2. §á?§Õ§Ò§à§â? §ä§Ñ§Ü§ä§Ú§Ü§Ú §Ý?§Ü§å§Ó§Ñ§ß§ß§ñ 3. §á§â§à§Ô§ß§à§Ù? §á§Ö§â§Ö§Ò?§Ô§å §Ù§Ñ§ç§Ó§à§â§ð§Ó§Ñ§ß§ß§ñ