FUTURES EXPO SERIES
POCT ASSAY FOR PRECISION DIAGNOSTICS
A streamlined CRISPR / Cas12a immunosensing platform : achieving attomolar sensitivity in small protein diagnostics
RESEARCH PROJECT
Recent years have seen significant progress in the field of CRISPR / Cas biosensing , with notable advancements in the sensitivity , specificity , and speed of nucleic acid detection .
> Awarded the NSSN Grand Challenge Fund ( RG230538 ). ●
> Developed CRISPR / Casassociated detection assays , methods , and kits , securing a 2023 PCT Patent with International Application No . PCT / AU2022 / 050829 .
However , the use of CRISPR / Cas biosensing systems for non-nucleic acid targets has remained limited . This study introduced a novel approach by synthesising a functional hybrid conjugate of an antibody and a single-stranded DNA oligonucleotide .
This synthesis successfully integrates CRISPR / Cas12abased signal amplification into various immunoassay schemes . As a result , they present a straightforward yet universally applicable method , termed the CRISPR-based Universal Immunoassay Signal Enhancer ( CRUISE ), which boosts conventional immunoassays by facilitating attomolar sensitivity in detecting small proteins .
CRUISE can be incorporated into various immunoassays through the primary or secondary antibody , offering sensitivity as low as 1 fg mL−1 and a six-log linear range for cytokine detection . This represents a 103-fold increase in sensitivity compared to commercial IFN-γ ELISA kits . The same 1 fg mL−1 sensitivity and six-log linear range were achieved for detecting IFN-γ in human plasma samples .
The researchers anticipate that CRUISE will offer a simple , user-friendly , and effective alternative for those relying on immunoassays , especially when facing sensitivity or detection ranges limitations .
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