Rapid simultaneous detection of multiple retroviral DNA sequences using the
polymerase chain reaction and capillary DNA chromatography
FJ Sunzeri, TH Lee, RG Brownlee and MP Busch
Irwin Memorial Blood Centers, San Francisco, CA 94118.
The polymerase chain reaction (PCR) technique is a powerful new tool for
amplifying target DNA, thus allowing for sensitive detection of specific
nucleic acid sequences. One important potential use of PCR involves
screening the donated blood supply for transfusion-transmitted viruses.
Realization of this goal has been limited by (1) the requirement for
multiple, discrete PCR reactions to amplify and detect target sequences of
more than one virus, and (2) the lack of a rapid, nonhazardous means for
specific detection of one or more PCR-amplified products. We report the
simultaneous amplification of three distinct target sequences without
discernable loss in sensitivity toward any single target sequence. We also
demonstrate very rapid separation and detection of PCR-amplified viral DNA
through the use of automated capillary DNA chromatography. Amplified DNA
peaks were initially identified by scanning the capillary effluent at
ultraviolet wavelengths, while discrimination of human immunodeficiency
virus type 1 and human T-cell leukemic virus type I PCR-amplified DNA was
accomplished through use of virus-specific, fluorescently labeled primers
and probes. These results indicate progress toward an automated system for
screening the blood supply for nucleic acid sequences of multiple
pathogens.
Volume 77,
Issue 4,
pp. 879-886,
02/15/1991
Copyright © 1991 by The American Society of Hematology
