Microfabricated Isothermal Eg-Fet Sensor For Lamp Mediated Crispr/Cas12a Detection Of Hepatitis C Virus

A highly sensitive and rapid platform for the detection of hepatitis C virus (HCV) has been developed by combining loop-mediated isothermal amplification (LAMP) with the use of clustered regularly interspaced short palindromic repeats of associated protein 12 (CRISPR/cas12a). An isothermal sensor which serves as an indium tin oxide (ITO) based extended gate field-effect transistor (EG-FET) and is equipped with a polymethyl methacrylate (PMMA) microwell, is used to detect the potential change on the sensing surface, induced by the LAMP/CRISPR reaction. The immobilized signal reporter, which is single-strand deoxyribonucleic acid (ssDNA) on the ITO sensing surface, is cleaved by the activated CRISPR/cas12a. This trans-cleavage occurs due to the highly specific base-pairing of a nucleic acid target to a complementary guide ribonucleic acid (gRNA). The result is a significant potential change within four minutes. The diagnostic platform that is established in this study can be applied to the detection of other virus/bacteria through the use of different pathogen- specific LAMP/CRISPR assays. Therefore, this technique provides a rapid yet high performance platform for the detection of label-free pathogens.