title: A novel fluorescent aptasensor based on silica nanoparticles, PicoGreen and exonuclease III as a signal amplification method for ultrasensitive detection of myoglobin.
abstract: Measurement of myoglobin (Mb) in human blood serum is of great interest for quick diagnosis of acute myocardial infarction (AMI). In this study, a novel fluorescent aptasensor was designed for ultrasensitive and selective detection of Mb, based on target-induced high fluorescence intensity, complementary strand of aptamer (CS), PicoGreen (PG) dye, exonuclease III (Exo III) and silica nanoparticles coated with streptavidin (SNPs-Streptavidin). The developed aptasensor obtains characteristics of SNPs as enhancers of fluorescence intensity, Exo III as an enzyme which selectively digests the 3'-end of double-stranded DNA (dsDNA), PG as a fluorescent dye which could selectively bind to dsDNA and high selectivity and sensitivity of aptamer (Apt) toward its target. In the absence of Mb, no free CS remains in the environment of SNPs-Streptavidin, resulting in a weak fluorescence emission. In the present of Mb, dsDNA-modified SNPs-Streptavidin complex forms, leading to a very strong fluorescence emission. The developed fluorescent aptasensor exhibited high specificity toward Mb with a limit of detection (LOD) as low as 52 pM. In addition, the designed fluorescent aptasensor was efficiently used to detect Mb in human serum.
authors.affiliation: Academic Center for Education, Culture and Research (ACECR)-Mashhad Branch, Mashhad, Iran; Pharmaceutical Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address: abnouskh@mums.ac.ir.
authors.forename: Khalil
authors.initials: K
authors.lastname: Abnous
authors.affiliation: Nanotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran; Research Institute of Sciences and New Technology, Mashhad, Iran.
authors.forename: Noor Mohammad
authors.initials: NM
authors.lastname: Danesh
authors.affiliation: Cardiovascular Research Center, Ghaem Hospital, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Ahmad
authors.initials: A
authors.lastname: Sarreshtehdar Emrani
authors.affiliation: Nanotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Ramezani
authors.affiliation: Targeted Drug Delivery Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address: taghdisihm@mums.ac.ir.
authors.forename: Seyed Mohammad
authors.initials: SM
authors.lastname: Taghdisi
mesh_terms: D052157:Aptamers, Nucleotide; D005240:Feasibility Studies; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D057230:Limit of Detection; D009211:Myoglobin / Q000032:analysis*; D053758:Nanoparticles
chemical_list: D052157:Aptamers, Nucleotide; D005456:Fluorescent Dyes; D009211:Myoglobin
keywords: Exonuclease III; Fluorescent aptasensor; Myoglobin; PicoGreen; Silica nanoparticles