title: Using MF-PCR to diagnose multiple defects from single cells: implications for PGD.
abstract: Single cell genetic analysis is generally performed using PCR and FISH. Until recently, FISH has been the method of choice. FISH however is expensive, has significant misdiagnosis rates, can result in interpretation difficulties and is labour intensive making it unsuitable for high throughput processing. Recently fluorescent PCR reliability has increased to levels at or surpassing FISH whilst maintaining low cost. However, PCR accuracy has been a concern due to allelic dropout. Multiplex PCR can now increase accuracy by using multiple markers for each chromosome to firstly provide diagnosis if markers fail and/or secondly confirm diagnosis. We compare a variety of diagnostic methods and demonstrate for the first time a multiplex PCR system providing simultaneous diagnosis and confirmation of the major aneuploidy chromosomes (21,18,13) and sex as well as DNA fingerprint in single cells. We also discuss the implications of using PCR for aneuploidy screening in preimplantation genetic diagnosis.
authors.affiliation: Australian Genome Research Facility, Gehrmann Laboratories, The University of Queensland, Brisbane, QLD 4072, Australia. ian.findlay@agrf.org.au
authors.forename: I
authors.initials: I
authors.lastname: Findlay
authors.forename: P L
authors.initials: PL
authors.lastname: Matthews
authors.forename: B K
authors.initials: BK
authors.lastname: Mulcahy
authors.forename: K
authors.initials: K
authors.lastname: Mitchelson
mesh_terms: D000782:Aneuploidy; D001757:Blastomeres / Q000166:cytology; D002875:Chromosomes / Q000235:genetics; D003550:Cystic Fibrosis / Q000175:diagnosis; D016172:DNA Fingerprinting / Q000379:methods*; D005260:Female; D005315:Fetal Diseases / Q000175:diagnosis; D005456:Fluorescent Dyes; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D007621:Karyotyping; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D019836:Preimplantation Diagnosis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019849:Sex Determination Processes; D014314:Trisomy
chemical_list: D005456:Fluorescent Dyes