title: Direct RNA detection without nucleic acid purification and PCR: Combining sandwich hybridization with signal amplification based on branched hybridization chain reaction.
abstract: We have developed a convenient, robust and low-cost RNA detection system suitable for high-throughput applications. This system uses a highly specific sandwich hybridization to capture target RNA directly onto solid support, followed by on-site signal amplification via 2-dimensional, branched hybridizing chain polymerization through toehold-mediated strand displacement reaction. The assay uses SYBR Green to detect targets at concentrations as low as 1 pM, without involving nucleic acid purification or any enzymatic reaction, using ordinary oligonucleotides without modification or labeling. The system was demonstrated in the detection of malaria RNA in blood and GAPDH gene expression in cell lysate.
authors.affiliation: Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, Beijing, China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, Beijing, China. Electronic address: zhizheng100@126.com.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Zheng
mesh_terms: D052160:Benzothiazoles; D002460:Cell Line; D003959:Diamines; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D016778:Malaria, Falciparum / Q000097:blood; D009693:Nucleic Acid Hybridization / Q000379:methods*; D009930:Organic Chemicals / Q000737:chemistry; D010963:Plasmodium falciparum / Q000235:genetics; D011804:Quinolines; D012313:RNA / Q000032:analysis* / Q000097:blood; D016053:RNA, Protozoan / Q000032:analysis / Q000097:blood
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; D016053:RNA, Protozoan; C098022:SYBR Green I; D012313:RNA
keywords: Hybridization chain reaction; SYBR Green I; Signal amplification; Toehold