Development of RNA microchip for pathogen and cancer direct detection

摘要: Development of a simple, specific, sensitive and rapid RNA microchip for detection Head Neck Cancer (HNC) mRNA, pathogenic bacteria dengue virus (DENV) is reported. By use nucleases polymerases specific RNAs are selectively labeled detected without separation, reverse transcription or polymerase chain reaction. This accomplished by designing Hybrid probes consisting DNA-2’-O-Me-RNA-DNA regions to target the interest. Upon hybridization with RNA, RNase H digestion used remove 3’RNA sequences which exposes template Klenow extension reporter molecules such as hapten fluorophore labels. novel fast (ca. 1 h time), selective individual in synthetic mixture total mixtures, single nucleotide polymorphisms (SNPs) discrimination up attomole level chemiluminescence lower femtomole gold nanoparticles (AuNPs) silver staining method. Using chemiluminescence, HNC biomarkers, VCAM1 IL8 specifically presence thousands other mRNAs cancer cell lines human colon interference. Furthermore, method highly shown DENV SNPs discrimination. Moreover, we report (ca 1hour), selective, multi-marker using AuNPs-silver on microchip. Streptavidin technology has potential analysis wide array field including infectious diseases diagnosis, viral infections, food safety, gene expression profiling detection. A simple NaOH extraction procedure was developed E. coli results both AuNPs staining. avoids commercial purification kits thus reducing cost. visual does not require electricity special equipment, therefore good candidate diagnostics minimum resources. INDEX WORDS: Microarray, microchip, nanoparticles, neck cancer, Pathogenic detection, Dengue, polymorphism, lysis DEVELOPMENT OF MICROCHIP FOR PATHOGEN AND CANCER DIRECT DETECTION