title: Molecular investigations of mitochondrial deletions: evaluating the usefulness of different genetic tests.
abstract: Deletions in mitochondrial DNA are a common cause of mitochondrial disorders. The molecular diagnosis of mtDNA deletions for years was based on Southern hybridization later replaced by PCR methods such as PCR with primers specific for a particular deletion (mainly the so-called common deletion of 4977 bp) and long PCR. In order to evaluate the usefulness of MLPA (Multiplex Ligation-dependent Probe Amplification) in molecular diagnosis of large scale mtDNA deletions we compare four diagnostic methods: Southern hybridization, PCR, long-PCR and MLPA in a group of 16 patients with suspected deletions. Analysis was performed on blood, muscle and in one case hepatic tissue DNA. The MLPA was not able to confirm all the deletions detected by PCR methods, but due to its relative ease of processing, minimal equipment, low costs and the additional possibility to detect frequent point mtDNA mutations in one assay it is worth considering as a screening method. We recommend to always confirm MLPA results by PCR methods.
authors: Institute of Genetics and Biotechnology, Faculty of Biology, University of Warsaw ul. Pawińskiego 5A, 02-106 Warsaw, Poland. kaska@igib.uw.edu.pl
authors: Katarzyna
authors: K
authors: Tońska
authors: Dorota
authors: D
authors: Piekutowska-Abramczuk
authors: Magdalena
authors: M
authors: Kaliszewska
authors: Paweł
authors: P
authors: Kowalski
authors: Anna
authors: A
authors: Tańska
authors: Ewa
authors: E
authors: Bartnik
authors: Ewa
authors: E
authors: Pronicka
authors: Małgorzata
authors: M
authors: Krajewska-Walasek
mesh_terms: D000293:Adolescent; D015139:Blotting, Southern / Q000379:methods; D002648:Child; D002675:Child, Preschool; D004272:DNA, Mitochondrial / Q000235:genetics*; D005260:Female; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D022521:Ligase Chain Reaction; D008297:Male; D028361:Mitochondrial Diseases / Q000175:diagnosis* / Q000235:genetics*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D011446:Prospective Studies; D012189:Retrospective Studies; D017384:Sequence Deletion; D055815:Young Adult
chemical_list: D004272:DNA, Mitochondrial