title: A comparison between real-time quantitative PCR and DNA hybridization for quantitation of male DNA following myoblast transplantation.
abstract: The transplantation of muscle precursor cells (myoblasts) is a potential therapy for Duchenne muscular dystrophy. A commonly used method to detect cell survival is quantitation of the Y chromosome following transplantation of male donor cells into female hosts. This article presents a direct comparison between real-time quantitative PCR (Q-PCR) and the DNA hybridization (slot-blot) technique for quantitation of Y chromosome DNA. Q-PCR has a significantly greater linear quantitation range and is up to 40-fold more sensitive at low concentrations of male DNA, detecting as little as 1 ng of male DNA in each female tibialis anterior (TA) muscle. At high male DNA concentrations, accurate quantitation by Q-PCR is 2.5 times higher than the maximum possible with slot-blot. In conclusion, Q-PCR has a higher dynamic range and is more efficient than slot-blot analysis for the detection of donor cell engraftment in a transsexual transplantation model.
authors.affiliation: Discipline of Microbiology, School of Biomedical and Chemical Sciences, The University of Western Australia, Perth, WA, Australia.
authors.forename: Erika
authors.initials: E
authors.lastname: Bosio
authors.forename: Tracey F
authors.initials: TF
authors.lastname: Lee-Pullen
authors.forename: Clayton T
authors.initials: CT
authors.lastname: Fragall
authors.forename: Manfred W
authors.initials: MW
authors.lastname: Beilharz
authors.forename: Alayne L
authors.initials: AL
authors.lastname: Bennett
authors.forename: Miranda D
authors.initials: MD
authors.lastname: Grounds
authors.forename: Stuart I
authors.initials: SI
authors.lastname: Hodgetts
authors.forename: Leanne M
authors.initials: LM
authors.lastname: Sammels
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000235:genetics; D017690:Cell Transplantation / Q000379:methods; D003199:Computer Systems; D004247:DNA / Q000032:analysis* / Q000235:genetics; D004195:Disease Models, Animal; D005260:Female; D006085:Graft Survival / Q000235:genetics*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018101:Mice, Inbred mdx; D018482:Muscle, Skeletal / Q000737:chemistry / Q000473:pathology / Q000503:physiopathology; D009137:Muscular Dystrophy, Animal / Q000473:pathology / Q000628:therapy; D032446:Myoblasts / Q000166:cytology / Q000502:physiology / Q000637:transplantation*; D009693:Nucleic Acid Hybridization / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D014998:Y Chromosome / Q000235:genetics
chemical_list: D004247:DNA