21 KB117 KB20 KB199820252005Kokalj-Vokač, NadjaStangler Herodež, ŠpelaZagradišnik, Borisštevilka:2letnik:526 stranistr. 105-110ISSN:1318-0207COBISSID:1968447URN:URN:NBN:SI:doc-I7Q3GT9EenSlovensko kemijsko društvoActa chimica slovenicaCharcot-Marie-Tooth diseaseCharcot-Marie-Toothova bolezenCitogenetska analizaCytogenetic analysisDNAgenske analizeMLPAMLPA method for PMP22 gene analysis|DNA copy number alterations are responsible for several categories of human diseases and syndromes. These changes can be detected by cytogenetic studies when there is involvement of several kilobases or megabases of DNA. Examination of sub-microscopic changes is possible by using short probes flanked by the same primer pairs. Multiplex ligation-dependent probe amplification (MLPA) is a simple, high resolution method by which not sample nucleic acids but probes added to the samples are amplified and quantified. Charcot-Marie-Tooth disease type 1A (CMT1A) is the most common inherited peripheral neuropathy with a prevalence of about 1/10000. By a gene dosage mechanism, commonly trisomic overexpression of PMP22 results in CMT1A whereas its monosomic underexpression causes hereditary neuropathy with liability to pressure palsies (HNPP). We applied MLPA to the PMP22 gene in order to develop an efficient and sensitive test for detecting these gene dosage changes. The method was used on 56 samples collected for a previous comparative study on routine methods for CMT1A diagnosis used in our laboratory. All diagnoses agreed with results from other methods. The MLPA PMP22 assay is a simple, fast and accurate screening test for molecular diagnosis of CMT1A and HNPPPri ljudeh so spremembe v številu kopij DNA odgovorne za mnoge vrste bolezni in sindromov. Te spremembe se lahko odkrijejo s citogenetskimi raziskavami, kadar imamo prisotnih nekaj kilobaz ali megabaz DNA. Preiskovanje sub-mikroskopskih sprememb je možno ob uporabi kratkih prob, sestavljenih z enim parom prajmerjev. Hkratno pomnoževanje od ligacije odvisnih sond (MLPA) je enostavna metoda visoke ločljivosti, pri katerih se ne pomnožujejo in določajo nukleinske kisline vzorcev, ampak probe, dodane k vzorcem. Charcot-Marie-Toth-ovo obolenje tipa 1A (CMT1A) je najpogostejše dedno živčno mišično obolenje z razširjenostjo 1/10000. Pri CMT1A je prisotek presežek gena PMP22, medtem ko je njegov primanjkljaj značilen za HNPP. V ta namen smo v okviru gena PMP22 uporabili MLPA metodo, da bi razvili čimbolj učinkovit in občutljiv test za odkrivanje njegovih sprememb. Uporabili smo 56 vzorcev, predhodno testiranih z obstoječimi rutinskimi metodami laboratorija v okviru CMT1A diagnostike. Dobljeni rezultati so se ujemali z rezultati preostalih metod, kar pomeni, da je MLPA PMP22 analiza zaneljiva in v primerjavi z drugimi metodami precej enostavna in hitra v okviru molekularne diagnostike CMT1A in HNPPTEXTznanstveno časopisjejournalsSlovenian National E-content AggregatorNational and University Library of SloveniaSlovensko kemijsko društvo