One-Tube Nested PCR Coupled with CRISPR-Cas12a for Ultrasensitive Nucleic Acid Testing.

Nucleic acid testing with high sensitivity and specificity is of great importance for accurate disease diagnostics. Here, we developed an in situ one-tube nucleic acid testing assay. In this assay, the target nucleic acid is captured using magnetic silica beads, avoiding an elution step, followed directly by the polymerase chain reaction (PCR) and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas12a detection.

CRISPR/Cas12a coupled with loop-mediated isothermal amplification and lateral flow assay for SARS-CoV-2 detection.

Point-of-care testing (POCT) is rapid, exhibits highly sensitive performance, can facilitate home self-testing and avoids cross-contamination. Herein, we developed a biosensor that combines Si-OH magnetic bead (MB)-based fast RNA extraction, reverse transcription-loop-mediated isothermal amplification (RT-LAMP), CRISPR-Cas12a, and lateral flow assay (LFA) for rapid detection of SARS-CoV-2 RNA within 1.5 h.

Rapid and highly sensitive colorimetric LAMP assay and integrated device for visual detection of monkeypox virus.

Background: The monkeypox virus (MPXV) is a linear double-stranded DNA virus with a large genome that causes tens of thousands of infections and hundreds of deaths in at least 40 countries and regions worldwide. Therefore, timely and accurate diagnostic testing could be an important measure to prevent the ongoing spread of MPXV and widespread epidemics.

Results: Here, we designed multiple sets of primers for the target region of MPXV for loop-mediated isothermal amplification (LAMP) detection and identified the optimal primer set.