A high resolution HLA sequence-specific primer PCR reaction was done in a 96 well microtiter
plate. Each well contains primers to a different HLA allele and the same primers to a sequence of
DNA to be used as an internal control
A negative control consists of distilled water left in an
open tube during the specimen processing. After PCR, the contents of each well were
electrophoresed and visualized on gel. Which finding automatically indicates a potentially
invalid reaction that requires repeat analysis?
A) Two different alleles for the HLA-A locus were amplified.
B) An allele identified by the high resolution procedure does not match the allele identified
by the low-resolution procedure.
C) An allele determined by PCR does not match the allele determined by classic serology.
D) No bands were visualized in the negative control.
E) In the well that an HLA-A gene amplified, the control sequence also amplified (i.e., two
bands were visible).
B
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