title: Visual detection of nucleic acids based on lateral flow biosensor and hybridization chain reaction amplification.
abstract: In this study, a new lateral flow nucleic acid biosensor (LFNAB) using hybridization chain reaction (HCR) for signal amplification was developed for visual detection of nucleic acids with high sensitivity and low cost. A "sandwich-type" detection strategy was employed in our design. The sandwich system of capture probe (CP)/target DNA/reporter probe (RP)-HCR complexes was fabricated as the sensing platform. As the initiator strand, reporter probe propagated a chain reaction of hybridization events between the two hairpin probes modified with biotin, and determined whether long nicked DNA polymers were formed. The biotin-labeled double-strand DNA polymers then introduced numerous Streptavidin (SA)-labeled gold nanoparticles (AuNPs) on the lateral flow device. The CP/target DNA/RP-HCR complexes were captured on the test zone by the specific reaction between anti-Fam monoclonal antibody (anti-Fam mAb) on the test zone and Fam of the complexes. The accumulation of AuNPs on the test zone of the biosensor enabled the visual detection of specific sequences. The detection limit of specific DNA was as low as 1.76pM, which was about 2 orders lower than that of the LFNAB without HCR amplification. And the detection limit of Salmonella was 3×103cfumL-1. In conclusion, this visual detection system, HCR-LFNAB, is suitable for non-specialist personnel and point-of-care (POC) diagnosis in low-resource settings.
authors.affiliation: Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.
authors.forename: Na
authors.initials: N
authors.lastname: Ying
authors.affiliation: The Fourth Hospital of Jilin University, China.
authors.forename: Chuanjing
authors.initials: C
authors.lastname: Ju
authors.affiliation: Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.
authors.forename: Zhongyi
authors.initials: Z
authors.lastname: Li
authors.affiliation: Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China. Electronic address: Liuws85952@163.com.
authors.forename: Wensen
authors.initials: W
authors.lastname: Liu
authors.affiliation: Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China. Electronic address: wanjiayu@hotmail.com.
authors.forename: Jiayu
authors.initials: J
authors.lastname: Wan
mesh_terms: D001483:Base Sequence; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D001710:Biotin / Q000737:chemistry; D004247:DNA / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D006046:Gold / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry; D009693:Nucleic Acid Hybridization; D012313:RNA / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D012329:RNA, Bacterial / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D019779:Salmonella enterica / Q000235:genetics; D019809:Streptavidin / Q000737:chemistry
chemical_list: D015342:DNA Probes; D004269:DNA, Bacterial; D012329:RNA, Bacterial; D012313:RNA; D001710:Biotin; D006046:Gold; D004247:DNA; D019809:Streptavidin
keywords: 16S rRNA; Hybridization chain reaction; Lateral flow nucleic acid biosensor; Point of care; Salmonella; Visual detection