Selection of DNA aptamers against Mycobacterium tuberculosis Ag85A, and its application in a graphene oxide-based fluorometric assay.
Najmeh AnsariKiarash GhazviniMohammad RamezaniMahin ShahdordizadehRezvan Yazdian-RobatiKhalil AbnousSeyed Mohammad TaghdisiPublished in: Mikrochimica acta (2017)
The Mycobacterium Ag85 complex is the major secretory protein of M. tuberculosis. It is a potential marker for early diagnosis of tuberculosis (TB). The authors have identified specific aptamers for Ag85A (FbpA) via protein SELEX using magnetic beads. After twelve rounds of selection, two aptamers (Apt8 and Apt22) were chosen from different groups, and their binding constants were determined by flow cytometry. Apt22 (labeled with Atto 647N) binds to FbpA with high affinity (Kd = 63 nM) and specificity. A rapid, sensitive, and low-cost fluorescent assay was designed based on the use of Apt22 and graphene oxide, with a limit of detection of 1.5 nM and an analytical range from 5 to 200 nM of FbpA. Graphical abstract Schematic illustration of graphene oxide-based aptasensor for fluorometric determination of FbpA.
Keyphrases
- mycobacterium tuberculosis
- quantum dots
- molecularly imprinted
- flow cytometry
- low cost
- nucleic acid
- photodynamic therapy
- pulmonary tuberculosis
- loop mediated isothermal amplification
- sensitive detection
- label free
- high throughput
- binding protein
- highly efficient
- solid phase extraction
- visible light
- protein protein
- circulating tumor
- emergency department
- single molecule
- computed tomography
- light emitting
- risk assessment
- cell free
- hepatitis c virus
- climate change
- living cells
- single cell
- adverse drug
- hiv infected
- drug induced