Fluorescent PCR: a new technique for PGD of sex and single-gene defects.
BACKGROUND
For single-cell diagnosis, particularly preimplantation genetic diagnosis to be successful four main criteria must be achieved: sensitivity, reliability, accuracy, and identification/elimination of contamination.


RESULTS
Fluorescent PCR achieves all four necessary criteria and, in addition, currently allows genes on up to nine chromosomes to be simultaneously investigated. Fluorescent PCR has high sensitivity (approximately 1000 x conventional analysis systems), high reliability (97%), and high accuracy (97%) rates for both sex and CF diagnosis in single somatic cells. The low detection threshold allows allelic dropout (one of the main causes of misdiagnosis) to be easily distinguished from PCR phenomena such as preferential amplification. High reliability (90%) and accuracy (97-100%) have been achieved in sex and CF diagnosis in human blastomeres. Fluorescent PCR can also be used to DNA fingerprint (STR profiling) single cells to identify the source/origin of the cell and determine if contamination has occurred.


CONCLUSIONS
Fluorescent PCR is therefore a suitable method for PGD.
Findlay
I
I
Institute of Epidemiology and Health Services Researh, University of Leeds, UK. ianf@epid.leeds.ac.uk.
Quirke
P
P
Hall
J
J
Rutherford
A
A
D000483:Alleles; D053523:Amelogenin; D016477:Artifacts; D001757:Blastomeres / Q000737:chemistry*; D003550:Cystic Fibrosis / Q000175:diagnosis / Q000196:embryology / Q000235:genetics / Q000517:prevention & control; D004247:DNA / Q000235:genetics*; D016172:DNA Fingerprinting / Q000379:methods; D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers; D003746:Dental Enamel Proteins / Q000235:genetics; D005260:Female; D005315:Fetal Diseases / Q000175:diagnosis* / Q000517:prevention & control; D005456:Fluorescent Dyes; D005470:Fluorometry; D030342:Genetic Diseases, Inborn / Q000175:diagnosis* / Q000196:embryology / Q000235:genetics / Q000517:prevention & control; D005819:Genetic Markers; D006579:Heterozygote; D006801:Humans; D008297:Male; D009061:Mouth Mucosa / Q000166:cytology; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D012680:Sensitivity and Specificity; D012743:Sex Preselection / Q000379:methods*
D053523:Amelogenin; D017931:DNA Primers; D003746:Dental Enamel Proteins; D005456:Fluorescent Dyes; D005819:Genetic Markers; D004247:DNA