title: Development of analytical method for ultrasensitive detection of salbutamol utilizing DNA labeled-immunoprobe.
abstract: Salbutamol (SAL) is known as quick-acting β-2 receptor agonist and its use in humans for pulmonary diseases and/or in animal feed is limited because of associated potential hazardous effects on health. Several techniques are available for the detection of SAL, but are expensive and time consuming. Here for the first time, a novel pre-assembled DNA immunoprobe-based immuno-PCR assay was developed to investigate the levels of SAL in human urine samples and compared the proposed immuno-PCR assay for the detection of SAL with that of direct competitive ELISA. Under the optimized conditions, the assay showed a linear range over seven orders of magnitude, whereas the limit of detection of SAL in buffer was found to be 21 fg/mL. Current immuno-PCR assay exhibited a 300-fold better detection limit for SAL as compared to one achieved with direct competitive ELISA using the same antibody. In conclusion, it has been found that the developed method is highly sensitive and simple method that can significantly enhance the detection sensitivity for small molecules. Moreover it can be modified and used for detecting other small molecules and/or chemicals that exist in the environment and are responsible for the environmental pollution.
authors.affiliation: Institute of Pharmacology, Toxicology and Biochemical Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
authors.forename: Yajing
authors.initials: Y
authors.lastname: Lei
authors.affiliation: Tongde Hospital of Zhejiang Province, Hangzhou 310012, China.
authors.forename: Xiaoyi
authors.initials: X
authors.lastname: Li
authors.affiliation: Institute of Pharmacology, Toxicology and Biochemical Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China; Faculty of Pharmaceutical Sciences, Government College University Faisalabad, Faisalabad 38000, Pakistan.
authors.forename: Muhammad Sajid Hamid
authors.initials: MS
authors.lastname: Akash
authors.affiliation: Institute of Pharmacology, Toxicology and Biochemical Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
authors.forename: Lifang
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Institute of Pharmacology, Toxicology and Biochemical Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
authors.forename: Xiaojing
authors.initials: X
authors.lastname: Tang
authors.affiliation: Institute of Public Health, School of Medicine, Zhejiang University, Hangzhou 310058, China.
authors.forename: Weixing
authors.initials: W
authors.lastname: Shi
authors.affiliation: Hangzhou EPIE Bio-Detection Technology Limited, Hangzhou 310051, China.
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Institute of Pharmacology, Toxicology and Biochemical Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: chenshuqing@zju.edu.cn.
authors.forename: Shuqing
authors.initials: S
authors.lastname: Chen
mesh_terms: D000420:Albuterol / Q000737:chemistry* / Q000652:urine; D000906:Antibodies / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D057230:Limit of Detection; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity
chemical_list: D000906:Antibodies; D004247:DNA; D000420:Albuterol
keywords: Analytical method; Competitive; Immuno-polymerase chain reaction; Immunoassay; Salbutamol