On-line coupling of polymerase chain reaction and capillary electrophoresis for automatic DNA typing and HIV-1 diagnosis.
We demonstrate an integrated on-line system with a fused-silica capillary as the microreactor for PCR and capillary gel electrophoresis with laser-induced fluorescence detection for DNA typing and disease diagnosis. Two applications have been investigated: the four short tandem repeat (STR) loci vWA, THO1, TPOX and CSF1PO (CTTv) for DNA typing, and DNA probe for human immunodeficiency virus (HIV-1) diagnosis. The CTTv are important loci in forensic and genetic linkage analysis. The PCR technique is a powerful tool in HIV research because it can detect the presence of the virus before any antibody response in the infected person. Thus it is important for early diagnosis. Multiplexed PCR in a fused-silica capillary, on-line injection, DNA denaturation and calibration based on a standard ladder have been successfully combined. Also, on-line liquid flow management, DNA separation and detection have been completely integrated.
Zhang
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Ames Laboratory-USDOE and Department of Chemistry, Iowa State University, 50011, USA.
Yeung
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ES
D001322:Autoanalysis; D004247:DNA / Q000032:analysis*; D004279:DNA, Viral / Q000032:analysis / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000379:methods*; D015683:Gene Products, gag / Q000032:analysis; D015658:HIV Infections / Q000175:diagnosis*; D015497:HIV-1; D006801:Humans; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction / Q000379:methods*
D004279:DNA, Viral; D015683:Gene Products, gag; D004247:DNA