DETECTION OF AMPLIFIED Y-CHROMOSOME-SPECIFIC SEQUENCE BY CAPILLARY ELECTROPHORESIS WITH LASER-INDUCED FLUORESCENCE

Objective: To develop a sensitive method for genetic diagnosis using capillary electrophoresis with laser-induced fluorescence. Design: Using male DNA diluted with female DNA as an example, ZFY gene from Y chromosome was amplified specifically by polymerase chain reaction (PCR) with fluorescence-labeled primers and detected by capillary electrophoresis with laser-induced fluorescence. Setting: Laser operating laboratory. Participants: Human male and female DNA were extracted from healthy human male and female subjects. Interventions: None. Main Outcome Measures: The concentration of human DNA was determined by using a spectrophotometer at an absorbance of 260 nm. Result: Deoxyribonucleic acid fragments amplified from a single copy of ZFY gene were detected by capillary electrophoresis with laser-induced fluorescence after 35 cycles of PCR amplification. Conclusion: This method is potentially applicable for rapid and sensitive detection of fetal Y chromosome DNA sequence in maternal circulation and of single-cell DNA diagnosis.