title: Sensitive and selective label-free alkaline phosphatase detection based on DNA hairpin probe.
abstract: Here, we present a label-free, simple and signal-on architecture for fluorescent alkaline phosphatase (ALP) biosensor utilizing an SYBR Green I (SG) assisted fluorescence amplification method. The strategy relies on the fact that ALP provides a significant barrier to lambda exonuclease (λ exo) activity by a dephosphorylating DNA hairpin probe (HP), and SG shows a considerable fluorescence intensity enhancement upon binding to double-stranded DNA (dsDNA) than single-stranded DNA (ssDNA). Our method is simple, sensitive and selective, which can also successfully detect the activity of ALP in complex biological fluids. The results have revealed that the method allows a specific and quantitative assay of the target ALP with a wide linear response range from 0.4 to 200 U/mL and a detection limit of 0.05 U/mL.
authors.affiliation: State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, PR China.
authors.forename: Ziwei
authors.initials: Z
authors.lastname: Zhao
authors.forename: Wenping
authors.initials: W
authors.lastname: Zhu
authors.forename: Zhen
authors.initials: Z
authors.lastname: Li
authors.forename: Jianhui
authors.initials: J
authors.lastname: Jiang
authors.forename: Guoli
authors.initials: G
authors.lastname: Shen
authors.forename: Ruqin
authors.initials: R
authors.lastname: Yu
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis* / Q000378:metabolism; D000818:Animals; D052160:Benzothiazoles; D015374:Biosensing Techniques; D002417:Cattle; D004247:DNA / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry*; D003959:Diamines; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D009930:Organic Chemicals / Q000737:chemistry*; D011804:Quinolines
chemical_list: D052160:Benzothiazoles; D015342:DNA Probes; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D004247:DNA; D000469:Alkaline Phosphatase