Point-of-Care Extraction-Free Sample Preparation for Recombinase Polymerase Amplification of Nucleic Acid Targets in Buccal Swabs or Blood.

ABSTRACT: Introduction: Recombinase polymerase amplification (RPA) amplifies nucleic acid targets quickly and with low-complexity instrumentation. Simple sample preparation is needed for RPA testing at the point of care (POC). We describe and evaluate sample preparation workflows that do not require nucleic acid extraction for detecting β-globin DNA via RPA in swabs and blood. Methods: Amplification of targets present in crude sample lysate is suitable for the POC, but inhibitors can pose challenges. We determined the tolerance of RPA to various chemical lytic agents and sample matrices. We compared sample preparation methods determining the relative limit of detection as a function of pooled swab and comparing the performance across the dynamic range of white blood cell (WBC) counts.

SUBMITTER: Wilkinson AF 

PROVIDER: S-EPMC12658628 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature