Direct detection of multiple point mutations in mitochondrial DNA

Autor:	L. J. C. Wong, Dinithi Senadheera
Rok vydání:	1997
Předmět:	Genetics COLD-PCR Mutation Mitochondrial DNA Point mutation Biochemistry (medical) Clinical Biochemistry Biology medicine.disease_cause Molecular biology Heteroplasmy law.invention law Multiplex polymerase chain reaction medicine Restriction fragment length polymorphism Polymerase chain reaction
Zdroj:	Clinical Chemistry. 43:1857-1861
ISSN:	1530-8561 0009-9147
DOI:	10.1093/clinchem/43.10.1857
Popis:	Mitochondrial defects can be caused by mutations in nuclear or mitochondrial DNA. Large deletion/duplication and point mutations are the two major types of mitochondrial DNA (mtDNA) mutations. Comprehensive molecular diagnosis requires the analysis of multiple point mutations. We developed an effective multiplex PCR/allele-specific oligonucleotide (ASO) method to simultaneously screen multiple point mutations in mtDNA. The system involved three pairs of primers to amplify mutation “hot spots” at tRNAleu(UUR), tRNAlys/ATPase, and ND4 regions, followed by detection of point mutations with ASO probes. Over 2000 specimens were analyzed and the results were compared with those from previous studies with the PCR/restriction fragment length polymorphism method. Our data demonstrate that the multiplex PCR/ASO method is much more sensitive in the detection of low mutant heteroplasmy. It is simple and cost effective, especially if a large number of samples are to be screened for multiple point mutations.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::76635a67c2e724acdc562bc336b0a462 https://doi.org/10.1093/clinchem/43.10.1857 Zobrazit plný text záznamu