posted on 2021-06-28, 16:03authored byChristopher
P. Mancuso, Zhi-Xiang Lu, Jason Qian, Sarah A. Boswell, Michael Springer
Quantitative diagnostics that are
rapid, inexpensive, sensitive,
robust, and field-deployable are needed to contain the spread of infectious
diseases and inform treatment strategies. While current gold-standard
techniques are highly sensitive and quantitative, they are slow and
require expensive equipment. Conversely, current rapid field-deployable
assays available provide essentially binary information about the
presence of the target analyte, not a quantitative measure of concentration.
Here, we report the development of a molecular diagnostic test [quantitative
recombinase polymerase amplification (qRPA)] that utilizes competitive
amplification during a recombinase polymerase amplification (RPA)
assay to provide semi-quantitative information on a target nucleic
acid. We demonstrate that qRPA can quantify DNA, RNA, and viral titers
in HIV and COVID-19 patient samples and that it is more robust to
environmental perturbations than traditional RPA. These features make
qRPA potentially useful for at-home testing to monitor the progress
of viral infections or other diseases.