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

Objective: To develop a sensitive method for genetic diagnosis using c apillary electrophoresis with laser-induced fluorescence. Design: Usin g male DNA diluted with female DNA as an example, ZFY gene from Y chro mosome was amplified specifically by polymerase chain reaction (PCR) w ith fluorescence-labeled primers and detected by capillary electrophor esis with laser-induced fluorescence. Setting: Laser operating laborat ory. Participants: Human male and female DNA were extracted from healt hy human male and female subjects. Interventions: None. Main Outcome M easures: The concentration of human DNA was determined by using a spec trophotometer at an absorbance of 260 nm. Result: Deoxyribonucleic aci d fragments amplified from a single copy of ZFY gene were detected by capillary electrophoresis with laser-induced fluorescence after 35 cyc les of PCR amplification. Conclusion: This method is potentially appli cable for rapid and sensitive detection of fetal Y chromosome DNA sequ ence in maternal circulation and of single-cell DNA diagnosis.