1 research outputs found
Nanoliter Multiplex PCR Arrays on a SlipChip
The SlipChip platform was tested to perform highthroughput nanoliter multiplex PCR. The advantages of
using the SlipChip platform for multiplex PCR include the
ability to preload arrays of dry primers, instrument-free
sample manipulation, small sample volume, and highthroughput capacity. The SlipChip was designed to preload one primer pair per reaction compartment and to
screen up to 384 different primer pairs with less than 30
nanoliters of sample per reaction compartment. Both a
40-well and a 384-well design of the SlipChip were tested
for multiplex PCR. In the geometries used here, the
sample ļ¬uid was spontaneously compartmentalized into
discrete volumes even before slipping of the two plates of
the SlipChip, but slipping introduced additional capabilities that made devices more robust and versatile. The
wells of this SlipChip were designed to overcome potential
problems associated with thermal expansion. By using
circular wells ļ¬lled with oil and overlapping them with
square wells ļ¬lled with the aqueous PCR mixture, a
droplet of aqueous PCR mixture was always surrounded
by the lubricating ļ¬uid. In this design, during heating and
thermal expansion, only oil was expelled from the compartment and leaking of the aqueous solution was prevented. Both 40-well and 384-well devices were found to
be free from cross-contamination, and end point ļ¬uorescence detection provided reliable readout. Multiple samples
could also be screened on the same SlipChip simultaneously. Multiplex PCR was validated on the 384-well
SlipChip with 20 different primer pairs to identify 16
bacterial and fungal species commonly presented in blood
infections. The SlipChip correctly identiļ¬ed ļ¬ve different
bacterial or fungal species in separate experiments. In
addition, the presence of the resistance gene mecA in
methicillin resistant Staphylococcus aureus (MRSA) was
identiļ¬ed. The SlipChip will be useful for applications
involving PCR arrays and lays the foundation for new
strategies for diagnostics, point-of-care devices, and immobilization-based arrays