DIAGNOSTIC-DETECTION OF HERPES-SIMPLEX AND HEPATITIS-C VIRAL AMPLICONS BY CAPILLARY ELECTROPHORESIS - COMPARISON WITH SOUTHERN BLOT DETECTION

Background: As a result of the large number of DNA-based clinical assa ys, there is intense interest in making polymerase chain reaction (PCR )-amplified DNA product analysis faster, more cost-effective, and more automated. Methods and Results: In this study, an evaluation of the u se of capillary gel electrophoresis with laser-induced fluorescence de tection is described as a means of analyzing postamplification PCR pro ducts from clinical specimens. Sixty-six herpes simplex virus and 152 hepatitis C virus amplicons were analyzed after PCR and reverse transc ription PCR, respectively. It is shown that the use of a physical gel buffer system in a short capillary in conjunction with laser-induced f luorescence detection allows for sensitive detection of herpes simplex virus-and hepatitis C virus-specific DNA fragments in an expedient ma nner. Interinstrument and intercapillary reproducibility of the migrat ion time was evaluated and found to be excellent. The advantages and d isadvantages over agarose gel electrophoresis-Southern blot analysis a re summarized. Conclusions: The advantages offered by capillary gel el ectrophoresis with laser-induced fluorescent detection including rapid and sensitive analysis, ease of setup, reduced cost, and possibility for automation, make this procedure a viable alternative to more labor -intensive agarose gel electrophoresis-Southern blot analysis as molec ular diagnostic methodology.