title: Self-assembly formed by a short DNA probe pair: Application for highly sensitive mRNA species detection without reverse transcription.
abstract: We describe a novel technology for detecting nucleic acids: Probe Alteration Link Self-Assembly Reactions (PALSAR). PALSAR comprises DNA self-assembly of pairs of short DNA probes formed by alternate hybridization of three complementary regions in a pair of honeycomb probes (HCPs). Self-assembly occurs at designated salt concentrations and reaction temperatures and requires no enzymes. We prepared pairs of HCPs to detect mRNAs encoded by the GAPDH gene β-actin (BA) gene, CD3D gene, CD4 gene, major vault protein (MV) gene and the signalling lymphocytic activation molecule-associated protein (SAP) gene, and succeeded in quantitatively detecting these mRNAs. PALSAR could detect mRNA directly without synthesizing cDNA. Moreover, multiple mRNAs could be detected simultaneously in a single reaction tube and there was a good correlation between the results obtained PALSAR and those by real-time PCR.
authors.affiliation: PALMA Bee'Z Research Institute Co., Ltd., Japan; Division of Hematology and Rheumatology, Department of Medicine, Nihon University School of Medicine, 30-1 Oyaguchi, Itabashi, Tokyo, 173-8610, Japan.
authors.forename: Mitsugu
authors.initials: M
authors.lastname: Usui
authors.affiliation: PALMA Bee'Z Research Institute Co., Ltd., Japan; EIDIA Co., Ltd, 3262-12 Yoshiwara, Ami, Inashiki, Ibaraki, 300-1155, Japan; Division of Hematology and Rheumatology, Department of Medicine, Nihon University School of Medicine, 30-1 Oyaguchi, Itabashi, Tokyo, 173-8610, Japan.
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Fujikawa
authors.affiliation: EIDIA Co., Ltd, 3262-12 Yoshiwara, Ami, Inashiki, Ibaraki, 300-1155, Japan; Division of Hematology and Rheumatology, Department of Medicine, Nihon University School of Medicine, 30-1 Oyaguchi, Itabashi, Tokyo, 173-8610, Japan.
authors.forename: Masako
authors.initials: M
authors.lastname: Osawa
authors.affiliation: PALMA Bee'Z Research Institute Co., Ltd., Japan.
authors.forename: Chikako
authors.initials: C
authors.lastname: Hakii
authors.affiliation: Division of Hematology and Rheumatology, Department of Medicine, Nihon University School of Medicine, 30-1 Oyaguchi, Itabashi, Tokyo, 173-8610, Japan.
authors.forename: Natsumi
authors.initials: N
authors.lastname: Ikumi
authors.affiliation: Division of Hematology and Rheumatology, Department of Medicine, Nihon University School of Medicine, 30-1 Oyaguchi, Itabashi, Tokyo, 173-8610, Japan.
authors.forename: Takamasa
authors.initials: T
authors.lastname: Nozaki
authors.affiliation: Division of Hematology and Rheumatology, Department of Medicine, Nihon University School of Medicine, 30-1 Oyaguchi, Itabashi, Tokyo, 173-8610, Japan.
authors.forename: Noboru
authors.initials: N
authors.lastname: Kitamura
authors.affiliation: Division of Hematology and Rheumatology, Department of Medicine, Nihon University School of Medicine, 30-1 Oyaguchi, Itabashi, Tokyo, 173-8610, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Hatta
authors.affiliation: PALMA Bee'Z Research Institute Co., Ltd., Japan; Department of Infectious Diseases, National Center for Child Health and Development, 2-10-1 Okura, Setagaya, Tokyo, 157-8535, Japan.
authors.forename: Shigeyoshi
authors.initials: S
authors.lastname: Fujiwara
authors.affiliation: Division of Hematology and Rheumatology, Department of Medicine, Nihon University School of Medicine, 30-1 Oyaguchi, Itabashi, Tokyo, 173-8610, Japan. Electronic address: takei.masami@nihon-u.ac.jp.
authors.forename: Masami
authors.initials: M
authors.lastname: Takei
mesh_terms: D015342:DNA Probes; D057230:Limit of Detection; D012333:RNA, Messenger / Q000235:genetics*; D048348:Reverse Transcription
chemical_list: D015342:DNA Probes; D012333:RNA, Messenger
keywords: Gene expression; Luminex; Multiplex detection; PALSAR; Self-assembly; Signal amplification