posted on 2023-04-28, 15:36authored byNan Rong, Kaiyue Chen, Jiqi Shao, Qi Ouyang, Chunxiong Luo
As an absolute quantification method
at the single-molecule
level,
digital PCR (dPCR) offers the highest accuracy. In this work, we developed
a 3D scalable chamber-array chip that multiplied the number of partitions
by stacking chamber-array layers and realized digital loop-mediated
isothermal amplification to quantify DNA molecules. It greatly increases
the number of partitions to improve the performance of dPCR without
increasing the chip size, the operation workflow complicity, and operation
time. For the three-chamber-array-layer chip which contains 200,000
reactors of a 0.125 nL volume, it has been proved that the reagent
filling and partition were finished within 3 min, and the whole detection
could be finished within 1 h. The method demonstrated that it could
be scalable to a six-chamber-array layer, which contains 400,000 reactors
without increasing the size of the chip and the complication of filling/partition
workflow but only takes an additional hour for scanning. Due to its
potential for high throughput, low cost, and simple operation, our
device may significantly expand the clinical application range of
dPCR.