To minimize and control the transmission of infectious diseases,a sensitive,accurate,rapid,and robust assay strategy for application on-site screening is critical.Here,we report single-molecule RNA capture-assisted di...To minimize and control the transmission of infectious diseases,a sensitive,accurate,rapid,and robust assay strategy for application on-site screening is critical.Here,we report single-molecule RNA capture-assisted digital RT-LAMP(SCADL)for point-of-care testing of infectious diseases.Target RNA was captured and enriched by specific capture probes and oligonucleotide probes conjugated to magnetic beads,replacing laborious RNA extraction.Droplet generation,amplification,and the recording of results are all integrated on a microfluidic chip.In assaying commercial standard samples,quantitative results precisely corresponded to the actual concentration of samples.This method provides a limit of detection of 10 copies mL−1 for the N gene within 1 h,greatly reducing the need for skilled personnel and precision instruments.The ultrasensitivity,specificity,portability,rapidity and user-friendliness make SCADL a competitive candidate for the on-site screening of infectious diseases.展开更多
基金supported by the National Natural Science Foundation of China(Nos.81902167,32071407,62003023,32101088,12072010,and 62073299)the National Key Research and Development Program of China(2022YFB3205600)+1 种基金the Beijing Natural Science Foundation(No.7212204)the Project of Central Plains Science and Technology Innovation Leading Talents(No.224200510026).
文摘To minimize and control the transmission of infectious diseases,a sensitive,accurate,rapid,and robust assay strategy for application on-site screening is critical.Here,we report single-molecule RNA capture-assisted digital RT-LAMP(SCADL)for point-of-care testing of infectious diseases.Target RNA was captured and enriched by specific capture probes and oligonucleotide probes conjugated to magnetic beads,replacing laborious RNA extraction.Droplet generation,amplification,and the recording of results are all integrated on a microfluidic chip.In assaying commercial standard samples,quantitative results precisely corresponded to the actual concentration of samples.This method provides a limit of detection of 10 copies mL−1 for the N gene within 1 h,greatly reducing the need for skilled personnel and precision instruments.The ultrasensitivity,specificity,portability,rapidity and user-friendliness make SCADL a competitive candidate for the on-site screening of infectious diseases.
