{"id":1129,"date":"2016-12-05T22:15:35","date_gmt":"2016-12-05T21:15:35","guid":{"rendered":"http:\/\/www.newslab.sk\/2016\/12\/05\/komparativna-genomova-hybridizacia-uvod-do-metodiky\/"},"modified":"2017-10-03T14:57:48","modified_gmt":"2017-10-03T12:57:48","slug":"comparative-genomic-hybridization-a-methodological-introduction","status":"publish","type":"post","link":"https:\/\/www.newslab.sk\/en\/comparative-genomic-hybridization-a-methodological-introduction\/","title":{"rendered":"Comparative genomic hybridization: a methodological introduction"},"content":{"rendered":"
*All tables, charts, graphs and pictures that are featured in this article can be found in the .pdf\r\n attachment at the end of the paper. <\/strong><\/span><\/pre>\n
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\u00davod<\/strong><\/p>\n
Incidencia zhubn\u00fdch n\u00e1dorov m\u00e1 celosvetovo st\u00fapaj\u00faci trend. Slovensko nie je v\u00fdnimka. N\u00e1rodn\u00fd onkologick\u00fd register SR za rok 2008 evidoval u\u00a0oboch pohlav\u00ed spolu 30\u00a0144 os\u00f4b s n\u00e1dorov\u00fdmi ochoreniami, z\u00a0toho 15\u00a0055\u00a0mu\u017eov a\u00a015\u00a0089 \u017eien.<\/p>\n
Zo \u0161tatist\u00edk pre rok 2008 vypl\u00fdva, \u017ee v\u00a0Slovenskej republike medzi naj\u010dastej\u0161ie sa vyskytuj\u00face n\u00e1dorov\u00e9 ochorenia u\u00a0\u017eien patria zhubn\u00e9 n\u00e1dory prsn\u00edka, tela maternice, kr\u010dka maternice a vaje\u010dn\u00edkov. Zhubn\u00e9 n\u00e1dory prsn\u00edkov tvoria po nemelan\u00f3mov\u00fdch n\u00e1doroch ko\u017ee druh\u00fd naj\u010dastej\u0161ie sa vyskytuj\u00faci typ n\u00e1dorov\u00e9ho ochorenia u\u00a0\u017eien. V\u00a0roku 2008 bolo hl\u00e1sen\u00fdch 2639 nov\u00fdch pr\u00edpadov s 772 \u00famrtiami. Zhubn\u00e9 n\u00e1dory tela maternice boli zaznamenan\u00e9 876 kr\u00e1t, pri\u010dom 221 postihnut\u00fdch n\u00e1sledkom tohto ochorenia pod\u013eahlo. Zhubn\u00e9 n\u00e1dory kr\u010dka maternice boli hl\u00e1sen\u00e9 u\u00a0634 pacientok, z \u010doho 209 kon\u010dilo exitom. Zhubn\u00e9 n\u00e1dory vaje\u010dn\u00edkov boli evidovan\u00e9 u\u00a0509 pacientok s\u00a0hl\u00e1sen\u00fdmi 276 \u00famrtiami (1).<\/p>\n
Onkologick\u00e9 ochorenia vo v\u0161eobecnosti pova\u017eujeme za ochorenie g\u00e9nov. T\u00fato skuto\u010dnos\u0165 najlep\u0161ie ilustruj\u00fa famili\u00e1rne viazan\u00e9 ochorenia, pri ktor\u00fdch sa muta\u010dn\u00e9 po\u0161kodenie konkr\u00e9tneho g\u00e9nu pren\u00e1\u0161a z\u00a0gener\u00e1cie na gener\u00e1ciu a u\u00a0postihnut\u00e9ho jedinca sa tak zvy\u0161uje riziko vypuknutia ochorenia v\u00a0skor\u0161\u00edch rokoch \u017eivota. So zvy\u0161uj\u00facou sa priemernou d\u013a\u017ekou \u017eivota je mo\u017en\u00e9 predpoklada\u0165 aj zvy\u0161uj\u00facu sa incidenciu mal\u00edgnych ochoren\u00ed. V\u00a0tomto smere bude st\u00fapa\u0165 potreba nov\u00fdch skr\u00edningov\u00fdch met\u00f3d umo\u017e\u0148uj\u00facich efekt\u00edvne a\u00a0v\u010dasn\u00e9 odhalenie ochorenia. Medzi tak\u00e9 patr\u00ed aj metodika naz\u00fdvan\u00e1 komparat\u00edvna gen\u00f3mov\u00e1 hybridiz\u00e1cia \u2013 CGH.<\/p>\n
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Skr\u00edningov\u00e9 metodiky cytogenetiky<\/strong><\/p>\n
Od\u00a0sedemdesiatich rokov minul\u00e9ho storo\u010dia sa na sledovanie nebalansovan\u00fdch chromoz\u00f3mov\u00fdch abnormal\u00edt, ako s\u00fa strata alebo zisk genetick\u00e9ho materi\u00e1lu, vyu\u017e\u00edvaj\u00fa r\u00f4zne druhy pr\u00fa\u017ekovania chromoz\u00f3mov. V\u00fdsledkom aplik\u00e1cie metodiky s\u00fa pod mikroskopom vidite\u013en\u00e9 pr\u00fa\u017eky na kondenzovan\u00fdch chromoz\u00f3moch, \u010do u\u013eah\u010duje identifik\u00e1ciu a\u00a0lokaliz\u00e1ciu abnormal\u00edt. Metodika dosahuje\u00a0v\u00a0ide\u00e1lnych pr\u00edpadoch rozl\u00ed\u0161enie 5Mb (2).<\/p>\n
Ku klasick\u00fdm metodik\u00e1m cytogenetiky patr\u00ed aj fluorescen\u010dn\u00e1 in situ hybridiz\u00e1cia (FISH). Fluorescen\u010dne zna\u010den\u00e9 pr\u00f3by, RNA alebo DNA, sa na z\u00e1klade komplementarity via\u017eu na presne zodpovedaj\u00facu sekvenciu nukleotidov chromoz\u00f3movej DNA. Pou\u017eit\u00edm fluorescen\u010dn\u00e9ho mikroskopu a\u00a0pr\u00edslu\u0161n\u00e9ho softv\u00e9ru je mo\u017en\u00e9 lokalizova\u0165 a detegova\u0165\u00a0pr\u00edtomnos\u0165 alebo nepr\u00edtomnos\u0165 konkr\u00e9tneho genetick\u00e9ho materi\u00e1lu v\u00a0danej oblasti chromoz\u00f3mov. Zaveden\u00edm \u201esuper-resolution\u201c techn\u00edk sa rozli\u0161ovacia schopnos\u0165 FISH metodiky zv\u00fd\u0161ila a\u017e na mo\u017enos\u0165 detekcie nieko\u013eko m\u00e1lo kilob\u00e1z (3).<\/p>\n
Niekam medzi klasick\u00e9 cytogenetick\u00e9 metodiky, ako je karyotypovanie a\u00a0FISH, a\u00a0molekul\u00e1rno-biologick\u00e9 techniky, ako je kvantitat\u00edvna polymer\u00e1zov\u00e1 re\u0165azov\u00e1 reakcia v\u00a0re\u00e1lnom \u010dase (qRT-PCR) \u010di sekvenovanie, zara\u010fujeme komparat\u00edvnu gen\u00f3mov\u00fa hybridiz\u00e1ciu (CGH) a jej modernej\u0161iu vetvu v podobe array CGH (aCGH).<\/p>\n
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Kr\u00e1tko z\u00a0hist\u00f3rie CGH<\/strong><\/p>\n
Komparat\u00edvna gen\u00f3mov\u00e1 hybridiz\u00e1cia, CGH, bola prv\u00fdkr\u00e1t pop\u00edsan\u00e1 v\u00a0roku 1992 skupinou vedcov z\u00a0kalifornskej univerzity (4). Metodiku zalo\u017eili na kompetit\u00edvnej fluorescen\u010dnej in situ hybridiz\u00e1cii. Metaf\u00e1zov\u00e9 chromoz\u00f3my na skl\u00ed\u010dku hybridizovali s odli\u0161ne fluorescen\u010dne zna\u010den\u00fdmi referen\u010dn\u00fdmi a\u00a0testovan\u00fdmi \u00fasekmi DNA. Hybridiz\u00e1ciu testovanej a\u00a0referen\u010dnej DNA sondy hodnotili pod\u013ea intenzity v\u00fdsledn\u00e9ho farebn\u00e9ho sign\u00e1lu. Oblasti chromoz\u00f3mov, ktor\u00e9 stratili, alebo naopak z\u00edskali genetick\u00fd materi\u00e1l, mali v\u00a0porovnan\u00ed s\u00a0norm\u00e1lnou referen\u010dnou vzorkou zmenen\u00fd pomer intenz\u00edt oboch sign\u00e1lov, \u010do sa prejavilo vo v\u00fdslednej farbe sign\u00e1lu (4).<\/p>\n
V\u00a0roku 1997 autorsk\u00fd kolekt\u00edv z Heidelbergu publikoval upraven\u00fd variant CGH. Namiesto hybridiz\u00e1cie DNA sond s\u00a0chromoz\u00f3mami pou\u017eili na matrix naviazan\u00e9 kr\u00e1tke \u00faseky DNA a\u00a0tie hybridizovali s\u00a0testovanou a\u00a0referen\u010dnou DNA. Metodiku tak zjednodu\u0161ili a zna\u010dne zv\u00fd\u0161ili jej rozli\u0161ovaciu schopnos\u0165 (5). Dnes technol\u00f3gia array CGH (alebo matrix CGH) dosahuje rozli\u0161ovaciu schopnos\u0165 na \u00farovni 100 kb (6). Publikovan\u00e1 metodika polo\u017eila z\u00e1klad automatiz\u00e1cii procesu CGH a\u00a0otvorila t\u00fdm dvere do rutinn\u00fdch diagnostick\u00fdch laborat\u00f3ri\u00ed.<\/p>\n
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Princ\u00edp aCGH<\/strong><\/p>\n
Oba typy metodiky CGH, konven\u010dn\u00e1 na chromoz\u00f3moch, aj array CGH na b\u00e1ze DNA, s\u00fa zalo\u017een\u00e9 na rovnakom princ\u00edpe (Obr. 1). Testovan\u00e1 a referen\u010dn\u00e1 DNA je \u0161pecificky fluorescen\u010dne zna\u010den\u00e1. Testovan\u00e1 DNA sa oby\u010dajne zna\u010d\u00ed \u010derven\u00fdm farbivom (Cyan\u00edn-5) a\u00a0referen\u010dn\u00e1 DNA zelen\u00fdm fluorof\u00f3rom (Cyan\u00edn-3). Obe DNA s\u00fa spolo\u010dne a v\u00a0rovnakom pomere nanesen\u00e9 na skl\u00ed\u010dko (DNA microarray), ktor\u00e9 obsahuje presne \u0161pecifikovan\u00e9 jednovl\u00e1knov\u00e9 \u00faseky DNA, tzv. sondy, alebo klony. Ich d\u013a\u017eka sa u\u00a0jednotliv\u00fdch v\u00fdrobcov l\u00ed\u0161i (napr. Agilent pon\u00faka 60 b\u00e1zov\u00e9 sondy).<\/p>\n
Hybridiz\u00e1cia prebieha pri presne regulovan\u00fdch podmienkach nieko\u013eko hod\u00edn (24-56h), naj\u010dastej\u0161ie v\u00a0poloautomatick\u00fdch hybridiza\u010dn\u00fdch peciach. Po hybridiz\u00e1cii je skl\u00ed\u010dko prenesen\u00e9 do po\u010d\u00edta\u010dom riaden\u00e9ho skenera, kde je obraz nasn\u00edman\u00fd a softv\u00e9rovo\u00a0vyhodnoten\u00fd. Pre ka\u017ed\u00fd g\u00e9n je vypo\u010d\u00edtan\u00fd normalizovan\u00fd pomer intenz\u00edt \u010derven\u00e9ho a\u00a0zelen\u00e9ho sign\u00e1lu, ktor\u00fd indikuje zmenu v\u00a0mno\u017estve genetick\u00e9ho materi\u00e1lu na jednotliv\u00fdch \u00fasekoch DNA (tzv. CNV \u2013 Copy Number Variation). Ak je normalizovan\u00fd pomer <1 hovor\u00edme o\u00a0strate genetick\u00e9ho materi\u00e1lu (t. j. del\u00e9cia), ak je, naopak, pomer\u00a0 >1, ide o\u00a0zisk genetick\u00e9ho materi\u00e1lu (t. j. amplifik\u00e1cia) (Obr. 2).<\/p>\n
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Siln\u00e9 a\u00a0slab\u00e9 str\u00e1nky aCGH<\/strong><\/p>\n
Ka\u017ed\u00e1 metodika m\u00e1 svoje v\u00fdhody aj nev\u00fdhody. Slabou str\u00e1nkou metodiky je neschopnos\u0165 odhali\u0165 balansovan\u00e9 alter\u00e1cie, ako s\u00fa reciprok\u00e9 translok\u00e1cie alebo inverzie. Tieto anom\u00e1lie nemenia po\u010det k\u00f3pi\u00ed v\u00a0gen\u00f3me, a\u00a0tak ost\u00e1vaj\u00fa pre CGH.<\/p>\n
Nedetegovate\u013en\u00e9 ost\u00e1vaj\u00fa aj mut\u00e1cie, ktor\u00e9 s\u00fa mimo detek\u010dn\u00fd rozsah metodiky. Zaveden\u00edm aCGH sa rozl\u00ed\u0161enie metodiky zv\u00fd\u0161ilo na teoretick\u00fa hodnotu 100 kb. Rozl\u00ed\u0161enie aCGH je prim\u00e1rne ur\u010den\u00e9 dvoma faktormi: d\u013a\u017ekou cie\u013eovej DNA a hustotou pokrytia gen\u00f3mu sondami. Z\u00a0toho vypl\u00fdva, \u017ee \u010d\u00edm s\u00fa cie\u013eov\u00e9 sondy DNA na matrixe krat\u0161ie a\u00a0\u010d\u00edm hustej\u0161ie je nimi pokryt\u00e1 dan\u00e1 oblas\u0165, t\u00fdm je rozl\u00ed\u0161enie vy\u0161\u0161ie (7).<\/p>\n
Medzi najv\u00e4\u010d\u0161ie prednosti aCGH patr\u00ed schopnos\u0165 simult\u00e1nnej detekcie aneuplo\u00eddi\u00ed, del\u00e9ci\u00ed, inzerci\u00ed a duplik\u00e1ci\u00ed v\u00a0r\u00e1mci cel\u00e9ho gen\u00f3mu. Vhodn\u00fdm navrhnut\u00edm prekr\u00fdvaj\u00facich sa DNA sond m\u00f4\u017ee by\u0165 ur\u010dit\u00e1 oblas\u0165 v\u00a0r\u00e1mci gen\u00f3mu pokryt\u00e1 na 100%. To m\u00f4\u017ee by\u0165 v\u00fdhodn\u00e9 pri skr\u00edningu \u0161pecifick\u00fdch oblast\u00ed chromoz\u00f3mov, ktor\u00e9 s\u00fa priamo dotknut\u00e9 pri konkr\u00e9tnych druhoch ochorenia. Technika aCGH tak sp\u00e1ja v\u00fdhody lokusovo \u0161pecifickej FISH anal\u00fdzy s\u00a0mo\u017enos\u0165ou skr\u00edningu cel\u00e9ho gen\u00f3mu.<\/p>\n
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aCGH v diagnostike<\/strong><\/p>\n
Metodika aCGH na\u0161la svoje uplatnenie v\u00a0prenat\u00e1lnej a postnat\u00e1lnej diagnostike, v preimplanta\u010dnom skr\u00edningu, ale aj v\u00a0diagnostike n\u00e1dorov\u00fdch ochoren\u00ed.<\/p>\n
Prenat\u00e1lna a\u00a0postnat\u00e1lna diagnostika sa zameriava na v\u00fdvinov\u00e9 a ment\u00e1lne poruchy zapr\u00ed\u010dinen\u00e9 aber\u00e1ciami genetick\u00e9ho materi\u00e1lu. Samotn\u00e1 anal\u00fdza sa vykon\u00e1va na vzork\u00e1ch plodovej vody,\u00a0choriov\u00fdch klkov, ale aj perif\u00e9rnej krvi embrya a\u00a0nesk\u00f4r novorodenca. Kan a\u00a0kol. (8) uv\u00e1dza, \u017ee pomocou aCGH bolo identifikovan\u00fdch 20% (44\/220) klinicky signifikantn\u00fdch CNV pri\u00a0abnorm\u00e1lnych, ultrazvukom detegovan\u00fdch n\u00e1lezoch. Z tohto mno\u017estva 21 patrilo aneuplo\u00eddi\u00e1m a 23 in\u00fdm chromoz\u00f3mov\u00fdm imbalanci\u00e1m. Zauj\u00edmavos\u0165ou je, \u017ee 3,2% vzoriek (7\/220) s\u00a0CNV bolo detegovan\u00fdch iba prostredn\u00edctvom aCGH, a nie pou\u017eit\u00edm konven\u010dn\u00fdch cytogenetick\u00fdch techn\u00edk ako G-banding a kvantitat\u00edvna fluorescen\u010dn\u00e1 PCR. Z\u00a0toho d\u00f4vodu navrhuj\u00fa nahradi\u0165 karyotyping za metodiku aCGH a\u00a0vyu\u017ei\u0165 ju ako vo\u013ebu prvostup\u0148ov\u00e9ho skr\u00edningu. Konven\u010dn\u00e1 cytogenetika by sa mala pou\u017ei\u0165 n\u00e1sledne pri vizualiz\u00e1cii klinicky signifikantn\u00fdch CNV (8). Napriek tomu, \u017ee pomocou metodiky aCGH nie je mo\u017en\u00e9 zisti\u0165 v\u0161etky druhy po\u0161kodenia genetick\u00e9ho materi\u00e1lu, je schopn\u00e1 odhali\u0165 pribli\u017ene dvakr\u00e1t viac chromoz\u00f3mov\u00fdch abnormal\u00edt ako G-pr\u00fa\u017ekovanie (7).<\/p>\n
Citlivos\u0165 metodiky aCGH dokumentuje jej vyu\u017eitie v\u00a0preimplanta\u010dnom skr\u00edningu, kde je k\u00a0dispoz\u00edcii len minim\u00e1lne mno\u017estvo materi\u00e1lu. Pomocou aCGH je mo\u017en\u00e9 odhali\u0165 chromoz\u00f3mov\u00e9 imbalancie v\u00a0rozsahu cca 1Mb z\u00a0jedin\u00e9ho lymfoblastu, fibroblastu alebo blastom\u00e9ry (9).<\/p>\n
Hoci aCGH hr\u00e1 nezastupite\u013en\u00fa \u00falohu najm\u00e4 pri skr\u00edningu aber\u00e1ci\u00ed sp\u00f4sobuj\u00facich v\u00fdvinov\u00e9 a\u00a0ment\u00e1lne poruchy, svoje miesto si na\u0161la aj pri skr\u00edningu n\u00e1dorov\u00fdch ochoren\u00ed. V tejto oblasti v\u0161ak skr\u00edning nie je v\u017edy jednoduch\u00fd. Heterogenita buniek sp\u00f4soben\u00e1 ich klon\u00e1lnou povahou, nedostatok materi\u00e1lu (biopsia), jeho zn\u00ed\u017een\u00e1 kvalita (FFPE blo\u010dky), spolu so samotn\u00fdm typom aber\u00e1cie (n\u00edzky po\u010det k\u00f3pi\u00ed, aber\u00e1cie na kr\u00e1tkom \u00faseku), m\u00f4\u017eu metodike aCGH sp\u00f4sobi\u0165 probl\u00e9my. Napriek tomu sa aCGH s \u00faspechom pou\u017e\u00edva pri detekcii genomick\u00fdch abnormal\u00edt v\u00a0hematologick\u00fdch ochoreniach ako s\u00fa CLL (chronick\u00e1 lymfatick\u00e1 leuk\u00e9mia), MDS (myelodysplastick\u00fd syndr\u00f3m), MM (mnohopo\u010detn\u00fd myel\u00f3m), ALL (ak\u00fatna lymfoblastick\u00e1 leuk\u00e9mia), AML (ak\u00fatna myeloidn\u00e1 leuk\u00e9mia) a CMML (chronick\u00e1 myelomonocyt\u00e1rna leuk\u00e9mia) (10).<\/p>\n
Technol\u00f3gia aCGH sa osved\u010dila pri charakterizovan\u00ed genetick\u00fdch abnormal\u00edt zodpovedn\u00fdch napr. za zhubn\u00fd n\u00e1dor mo\u010dov\u00e9ho mech\u00fara (11), kolorekt\u00e1lnych n\u00e1dorov (12), n\u00e1dorov p\u013e\u00fac (13), pri identifik\u00e1cii nov\u00fdch g\u00e9nov zodpovedn\u00fdch za vznik n\u00e1dorov prostaty (15) a\u00a0mnoh\u00fdch \u010fal\u0161\u00edch.<\/p>\n
Metodika aCGH hr\u00e1 svoju \u00falohu aj pri \u0161t\u00fadiu naj\u010dastej\u0161ie sa vyskytuj\u00facich n\u00e1dorov\u00fdch ochoren\u00ed \u017eien. Ako je zn\u00e1me, n\u00e1dory prsn\u00edka s\u00fa heterog\u00e9nnou skupinou, v\u00a0ktorej etiol\u00f3gii nach\u00e1dzame mut\u00e1cie g\u00e9nov ako napr. BRCA1, BRCA2<\/em>, ale aj zmeny v\u00a0po\u010dte g\u00e9nov LSM1, BAG4 A\u00a0C8ORF4<\/em> pozorovan\u00e9 pomocou aCGH (15). Franc\u00fazska \u0161t\u00fadia v\u00a018 medic\u00ednskych centr\u00e1ch sledovala 423 pacientov s\u00a0metastatick\u00fdmi n\u00e1dormi, pri\u010dom pou\u017e\u00edvali Sangerovo sekvenovanie a\u00a0aCGH. Genomick\u00e9 alter\u00e1cie odhalili u\u00a0195 (46%) pacientov. Naj\u010dastej\u0161ie i\u0161lo o\u00a0aber\u00e1cie g\u00e9nov PIK3CA, CCND1<\/em> a\u00a0FGFR1<\/em>, ale aj zriedkav\u00e9 mut\u00e1cie v\u00a0g\u00e9ne AKT1 a zriedkav\u00e9\u00a0amplifik\u00e1cie v g\u00e9noch EGFR<\/em>, MDM2, FGFR2, AKT2, IGF1R a\u00a0MET.<\/em> Vybran\u00ed pacienti (55 zo 423) boli podroben\u00ed personalizovanej lie\u010dbe s\u00a0r\u00f4znym v\u00fdsledkom (16).<\/p>\n
N\u00e1dory vaje\u010dn\u00edkov s\u00fa taktie\u017e spojen\u00e9 s\u00a0muta\u010dn\u00fdm vyraden\u00edm tumor supresorov\u00fdch g\u00e9nov BRCA1\/2<\/em>, ale vyskytuj\u00fa sa aj ako s\u00fa\u010das\u0165 Li-Fraumeniho syndr\u00f3mu, v ktorom d\u00f4le\u017eit\u00fa \u00falohu hr\u00e1 g\u00e9n TP53<\/em>. \u0160pecifick\u00e9 genetick\u00e9 alter\u00e1cie boli pomocou aCGH odhalen\u00e9 aj v\u00a0pr\u00edpade ovari\u00e1lnych n\u00e1dorov, ktor\u00e9 vykazovali rezistenciu na chemoterapiu (17). Metodikou aCGH bol realizovan\u00fd skr\u00edning vybran\u00fdch onkog\u00e9nov z\u00a0endometri\u00e1lnych n\u00e1dorov, pri\u010dom bola pozorovan\u00e1 amplifik\u00e1cia onkog\u00e9nov AR, PIK3CA, MET, HRAS, NRAS, D17S1670, FGFR1, CTSB, RPS6KB1, LAMC2, MYC, PDGFRA, FGF4\/FGF3, PAKI <\/em>a\u00a0FGR<\/em> (18). Metodika aCGH zachytila CNV v\u00a073% (19\/26) vzoriek aj v\u00a0pr\u00edpade cervik\u00e1lnych n\u00e1dorov. Amplifik\u00e1cia bola zisten\u00e1 najm\u00e4 v oblastiach 3q (50,0%), 1q (42,4%), 19q (23,1%), k\u00fdm strata v\u00a0oblastiach 11q (30,8%), 4q (23,1%) a 13q (19,2%). Zv\u00fd\u0161en\u00fd v\u00fdskyt amplifik\u00e1cie 3q bol pozorovan\u00fd v\u00a0pozit\u00edvnych vzork\u00e1ch na HPV 16 a\u00a0HPV 18, \u010do len potvrdzuje z\u00e1va\u017enos\u0165 infekcie HPV pri tomto n\u00e1dorovom ochoren\u00ed (19).<\/p>\n
 <\/p>\n
Z\u00e1ver<\/strong><\/p>\n
Onkologick\u00e9 ochorenia aj napriek dok\u00e1zate\u013en\u00fdm \u00faspechom v\u00a0diagnostike a\u00a0lie\u010dbe predstavuj\u00fa v\u00e1\u017eny probl\u00e9m nielen z\u00a0medic\u00ednskeho h\u013eadiska. So zvy\u0161uj\u00facim sa priemern\u00fdm vekom popul\u00e1cie m\u00f4\u017eeme predpoklada\u0165, \u017ee incidencia t\u00fdchto ochoren\u00ed bude aj na\u010falej st\u00fapa\u0165. Z\u00a0toho d\u00f4vodu je \u017eiaduce, aby klinick\u00e9 laborat\u00f3ri\u00e1 mali k\u00a0dispoz\u00edcii modern\u00e9 metodiky, ktor\u00e9 umo\u017e\u0148uj\u00fa odhali\u0165 ochorenie na z\u00e1kladnej, molekulovej \u00farovni. Komparat\u00edvna gen\u00f3mov\u00e1 hybridiz\u00e1cia tieto podmienky sp\u013a\u0148a.<\/p>\n
Hoci pou\u017eitie aCGH v\u00a0skr\u00edningu onkologick\u00fdch ochoren\u00ed je v\u00a0niektor\u00fdch pr\u00edpadoch komplikovan\u00e9, v\u00fdsledky z\u00edskan\u00e9 touto metodikou maj\u00fa vysok\u00fa v\u00fdpovedn\u00fa hodnotu a\u00a0pravdepodobne bud\u00fa postupne nahr\u00e1dza\u0165 metodiky klasickej cytogenetiky. Odpor\u00fa\u010dania medzin\u00e1rodn\u00e9ho konzorcia pre \u0161tandardiz\u00e1ciu\u00a0 cytogenomick\u00fdch arra\u00ed – ISCA (International Standard Cytogenomic Array) pou\u017ei\u0165 CGH ako prv\u00fa skr\u00edningov\u00fa met\u00f3du a G-banding pou\u017e\u00edva\u0165 iba v\u00a0pr\u00edpadoch ako s\u00fa Downov syndr\u00f3m a\u00a0v\u00a0pr\u00edpadoch rodinn\u00e9ho v\u00fdskytu chromoz\u00f3mov\u00fdch aber\u00e1ci\u00ed (20) je len krokom k\u00a0tomuto prechodu. V\u00fdsledkom zavedenia nov\u00fdch technol\u00f3gi\u00ed do diagnostiky a\u00a0skr\u00edningu n\u00e1dorov\u00fdch ochoren\u00ed by mal by\u0165 genetick\u00fd profil pacienta, ktor\u00fd v\u00a0kone\u010dnom d\u00f4sledku vy\u00fasti do personalizovanej terapie.<\/p>\n
Projekt ITMS 26210120026 je spolufinancovan\u00fd zo zdrojov ES.<\/p>\n
 <\/p>\n
Literat\u00fara<\/strong><\/p>\n
    \n
  1. Safaei D, Plesko Incidencia zhubn\u00fdch n\u00e1dorov v\u00a0Slovenskej republike 2008. N\u00e1rodn\u00fd onkologick\u00fd register SR. NCZI; 2014.<\/li>\n
  2. Theisen A. Microarray-based comparative genomic hybridization (aCGH). Nature Education <\/em>2008;1:45.<\/li>\n
  3. Han R, Li Z, Fan Y, Jiang Y. Recent advances in super-resolution fluorescence imaging and its applications in biology. J Genet Genomics<\/em>. 2013;40(12):583-595.<\/li>\n
  4. Kallioniemi A, Kallioniemi OP, Sudar D, et al. Comparative genomic hybridization for molecular cytogenetic analysis of solid tumors. 1992;258(5083):818-821.<\/li>\n
  5. Solinas-Toldo S, Lampel S, Stilgenbauer S, et al. Matrix-based comparative genomic hybridization: Biochips to screen for genomic imbalances. Genes Chromosomes Cancer. <\/em>1997;20(4):399-407.<\/li>\n
  6. Friedman JM, Baross A, Delaney AD, et al. Oligonucleotide microarray analysis of genomic imbalance in children with mental retardation. Am J Hum Genet.<\/em> 2006;79(3):500-513.<\/li>\n
  7. Bejjani BA, Shaffer LG. Application of Array-Based Comparative Genomic Hybridization to Clinical Diagnostics. J Mol Diagn.<\/em> 2006;8(5):528-533.<\/li>\n
  8. Kan AS, Lau ET, Tang WF et al. Whole-genome array CGH evaluation for replacing prenatal karyotyping in Hong Kong. PLoS One.<\/em> 2014;9(2):e87988. doi: 10.1371\/journal.pone.0087988. eCollection 2014.<\/li>\n
  9. Greco E, Bono S, Ruberti A, et al. Comparative genomic hybridization selection of blastocysts for repeated implantation failure treatment: a pilot study. Biomed Res Int.<\/em> 2014; 2014:457913. doi: 10.1155\/2014\/457913. Epub 2014 Mar 23.<\/li>\n
  10. Simons A, Sikkema-Raddatz B, de Leeuw N, et al. Genome-wide arrays in routine diagnostics of hematological malignancies. Hum Mutat.<\/em> 2012;33(6):941-948.<\/li>\n
  11. Conconi D, Panzeri E, Redaelli S, et al. Chromosomal imbalances in human bladder urothelial carcinoma: similarities and differences between biopsy samples and cancer stem-like cells. BMC Cancer.<\/em> 2014;14:646. http:\/\/www.biomedcentral.com\/1471-2407\/14\/646<\/u><\/li>\n
  12. Brim H, Lee E, Abu-Asab MS, et al. Genomic aberrations in an African American colorectal cancer cohort reveals a MSI-specific profile and chromosome X amplification in male patients. PLoS One.<\/em> 2012;7(8):e40392. doi:10.1371\/journal.pone.0040392<\/li>\n
  13. Lo FY, Chang JW, Chang IS, et al. The database of chromosome imbalance regions and genes resided in lung cancer from Asian and Caucasian identified by array-comparative genomic hybridization. BMC Cancer.<\/em> 2012;12:235. http:\/\/www.biomedcentral.com\/1471-2407\/12\/235<\/u><\/li>\n
  14. Kamradt J, Jung V, Wahrheit K, et al. Detection of novel amplicons in prostate cancer by comprehensive genomic profiling of prostate cancer cell lines using oligonucleotide-based arrayCGH. PLoS One.<\/em> 2007;2(8):e769. doi: 1371\/journal.pone.0000769<\/li>\n
  15. Yang ZQ, Streicher KL, Ray ME, et al. Multiple interacting oncogenes on the 8p11-p12 amplicon in human breast cancer. Cancer Res.<\/em> 2006;66(24):11632-11643.<\/li>\n
  16. Andr\u00e9 F, Bachelot T, Commo F, et al. Comparative genomic hybridisation array and DNA sequencing to direct treatment of metastatic breast cancer: a multicentre, prospective trial (SAFIR01\/UNICANCER). Lancet Oncol.<\/em> 2014;15(3):267-274.<\/li>\n
  17. Osterberg L, Levan K, Parheen K, et al. Specific copy number alterations associated with docetaxel\/carboplatin response in ovarian carcinomas. Anticancer Res.<\/em> 2010;30(11):4451-4458.<\/li>\n
  18. O’Toole SA, Dunn E, Sheppard BL, et al. Genome-wide analysis of deoxyribonucleic acid in endometrial cancer using comparative genomic hybridization microarrays. Int J Gynecol Cancer.<\/em> 2006;16(2):834-842.<\/li>\n
  19. Kuglik P, Smetana J, Vallova V, et al. Genome-wide screening of DNA copy number alterations in cervical carcinoma patients with CGH+SNP microarrays and HPV-FISH. Int J Clin Exp Pathol.<\/em> 2014;7(8):5071-5082.<\/li>\n
  20. Coppinger J, Alliman S, Lamb AN, et al. Whole-genome microarray analysis in prenatal specimens identifies clinically significant chromosome alterations without increase in results of unclear significance compared to targeted microarray. Prenatal Diagnosis.<\/em> 2009;29(12):1156\u20131166.<\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"
    *All tables, charts, graphs and pictures that are featured in this article can be found in the .pdf attachment at the end of the paper.   \u00davod Incidencia zhubn\u00fdch n\u00e1dorov m\u00e1 celosvetovo st\u00fapaj\u00faci trend. Slovensko nie je v\u00fdnimka. N\u00e1rodn\u00fd onkologick\u00fd register SR za rok 2008 evidoval u\u00a0oboch pohlav\u00ed spolu 30\u00a0144 os\u00f4b s n\u00e1dorov\u00fdmi ochoreniami, z\u00a0toho<\/p>\n","protected":false},"author":7,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_mi_skip_tracking":false,"footnotes":""},"categories":[290],"tags":[730,731,371,616,732,733,572],"class_list":["post-1129","post","type-post","status-publish","format-standard","hentry","category-genetics","tag-acgh","tag-array-cgh","tag-breast-cancer-en","tag-cervical-cancer","tag-comparative-genomic-hybridization","tag-endometrial-cancer","tag-ovarian-cancer","typ_clanku-review-article"],"acf":{"abstrakt":"
    Traditional cytogenetic methods like banding or fluorescent in-situ hybridization have found a stable place in screening for genetic abnormalities as well as in evaluation of treatment response or prognosis. However, the development in the field of molecular biology over the past years has brought new methods, which enable assessment of genetic alterations at the molecular level, even simultaneously on the whole genome. Comparative genomic hybridization (CGH) belongs among such methods. The following article provides a brief introduction of the method, describes its principles, limitations as well as its application in diagnosing different kinds of diseases.<\/p>\n
    Key words: <\/strong>aCGH, array CGH, comparative genomic hybridization, breast cancer, endometrial cancer, cervical cancer, ovarian cancer<\/p>\n","casopis":[{"ID":1000,"post_author":"7","post_date":"2015-11-21 14:33:24","post_date_gmt":"2015-11-21 13:33:24","post_content":"
    GENETICS<\/strong><\/h4>\r\n
      \r\n \t
    • The laboratory age<\/li>\r\n \t
    • The Odyssey of DNA reading<\/li>\r\n \t
    • Comparative genomic hybridisation: a methodological introduction<\/li>\r\n \t
    • Next generation sequencing and its application in clinical genetics<\/li>\r\n<\/ul>\r\n
      <\/h4>\r\n
      BIOCHEMISTRY<\/strong><\/h4>\r\n
        \r\n \t
      • Determining of the trace elements in blood serum<\/li>\r\n \t
      • Determining of \u03b1-tocopherol (vitamin E) in serum by way of liquid chromatography with tandem mass spectrometry (LC\/MS\/MS)<\/li>\r\n \t
      • Analysis of urinary calculi and its path to Europe<\/li>\r\n<\/ul>\r\n \r\n
        IMMUNOLOGY <\/strong><\/h4>\r\n
          \r\n \t
        • New autoantibodies in diagnosis of autoimmunity myopathies<\/li>\r\n \t
        • Taking advantage of flow-based cytometry in determining prognostic markers<\/li>\r\n<\/ul>\r\nCD38 and ZAP-70 in patients with B-CLL\r\n\r\n \r\n
          CYTOLOGY AND PATHOLOGY <\/strong><\/h4>\r\n
            \r\n \t
          • Mucinous ovarian carcinoma \u2013 news in diagnosis from the pathologist\u2019s perspective<\/li>\r\n<\/ul>","post_title":"newsLab","post_excerpt":"","post_status":"publish","comment_status":"closed","ping_status":"closed","post_password":"","post_name":"newslab-12015","to_ping":"","pinged":"","post_modified":"2017-08-16 21:43:10","post_modified_gmt":"2017-08-16 19:43:10","post_content_filtered":"","post_parent":0,"guid":"http:\/\/www.newslab.sk\/casopis\/newslab-12015\/","menu_order":0,"post_type":"casopis","post_mime_type":"","comment_count":"0","filter":"raw"}],"strana":"11","upload_clanok":{"ID":1130,"id":1130,"title":"medirex_1_2015-tomka","filename":"MEDIREX_1_2015-\u2013-Tomka.pdf","filesize":287589,"url":"https:\/\/www.newslab.sk\/wp-content\/uploads\/2016\/12\/MEDIREX_1_2015-\u2013-Tomka.pdf","link":"https:\/\/www.newslab.sk\/en\/comparative-genomic-hybridization-a-methodological-introduction\/medirex_1_2015-tomka\/","alt":"","author":"7","description":"","caption":"","name":"medirex_1_2015-tomka","status":"inherit","uploaded_to":1129,"date":"2016-12-05 21:02:17","modified":"2016-12-05 21:02:17","menu_order":0,"mime_type":"application\/pdf","type":"application","subtype":"pdf","icon":"https:\/\/www.newslab.sk\/wp-includes\/images\/media\/document.png"}},"_links":{"self":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts\/1129","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/users\/7"}],"replies":[{"embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/comments?post=1129"}],"version-history":[{"count":0,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts\/1129\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/media?parent=1129"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/categories?post=1129"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/tags?post=1129"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}