AN EFFICIENT TECHNIQUE FOR DIRECT DENGUE VIRUS GENE DETECTION BASED ON IMMOBILIZATION OF QUANTUM DOTS VIA CLICK CHEMISTRY

Abstract

DENGUE FEVER IS A WORLDWIDE MOSQUITO-BORN DISEASE. ONE OF THE BIG PROBLEMS OF DENGUE FEVER IS THE DIFFICULTY TO IDENTIFY BY SYMPTOMS, SINCE DENGUE FEVER?S COMMON SYMPTOMS ARE SIMILAR TO OTHER FEBRILE ILLNESS. LABORATORY DIAGNOSIS IS NEEDED TO DETECT DENGUE VIRUS, AND PCR (POLYMERASE CHAIN REACTION) HAS BECOME AN IMPORTANT AND RELIABLE WAY TO DETECT AND IDENTIFY DENGUE VIRUS. IN THIS RESEARCH, WE COMBINED THE SALIENT OPTICAL PROPERTIES OF THE QUANTUM DOTS, CLICK CHEMISTRY, BIO-ORTHOGONAL LABELING AND PCR AMPLIFICATION STRATEGY FOR GENE DETECTION. IN THIS DETECTION SCHEME, DNA TEMPLATES ARE AMPLIFIED AND MODIFIED WITH BIOTIN ON ONE END AND AZIDE ON THE OTHER END BY PCR. STREPTAVIDIN (SA)-COATED QDS WERE ALSO CONJUGATED WITH THE ORTHOGONALLY-LABELLED AMPLICONS THROUGH AVIDIN-BIOTIN (AV/BT) CONJUGATION. THE QD CONJUGATES ARE THEN IMMOBILIZED ON A POLYSTYRENE BEADS VIA CLICK CHEMISTRY, THEREBY GENERATING A STRONG FLUORESCENCE SIGNAL. THIS DETECTION SCHEME MIGHT BE APPLIED TO POC MEDICAL DIA