Author/Editor		Stangler-Herodež, Špela; Zagradišnik, Boris; Kokalj-Vokač, Nadja
Title		MLPA method for PMP22 gene analysis
Type		članek
Source		Acta Chim Slov
Vol. and No.		Letnik 52, št. 2
Publication year		2005
Volume		str. 105-10
Language		eng
Abstract		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 amplication (MLPA) is a simple, high resolution method by which not sample nucleic acids 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 tdetecting 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 in a simple, fast and accurate screening test for molecular diagnosis of CMT1A and HNPP.
Summary		Pri Ijudeh 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 kateri se ne pomnožujejo in določajo nukleinske kisline vzorcev, ampak probe dodane k vzorcem. Charcot-Marie-Tooth-ovo obolenje tipa 1A (CMT1A) je najpogostejše dedno živčno mišično obolenje z razširjenostjo 1/10000. Pri CMT1A je prisoten presežek gena PMP22, medtem ko je njegov primanklaj značilen za HNPP. V ta nemen, 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 zanesljiva in v primerjavi z drugimi metodami precej enostavna in hitra v okviru molekularne diagnostike CMT1A in HNPP.
Descriptors		CHARCOT-MARIE DISEASE DNA PROBES GENE DOSAGE TRISOMY POLYMERASE CHAIN REACTION IN SITU HYBRIDIZATION, FLUORESCENCE