Detection of beta-glucans using an amperometric biosensor based on high-affinity interaction between Dectin-1 and beta-glucans.
Early diagnosis of fungal infection plays an important role in increasing antifungal therapeutic response, but meaningful tests such as microbiological cultures and histopathological diagnosis are usually insensitive and time-consuming. A sensitive amperometric biosensor for beta-glucans was fabricated by immobilizing Dectin-1 onto Nafion-thionine-gold nanoparticle-chitosan multilayer films to trap its corresponding ligand from sample solution. On formation of ligand-receptor complex, detection of beta-glucans was accomplished by monitoring the decrease of the electrochemical signal of the modified electrode due to the inhibition of the transmission of electrons. Dectin-1 was constructed by cloning the extracellular carbohydrate recognition domain of the mouse Dectin gene into the pET28a(+) prokaryotic expression vector. Optimal conditions and analytical performances of the described biosensor were investigated. Under the optimal conditions, the biosensor response for beta-glucans presented good accuracy, stability, and reproducibility. The proposed biosensor not only could be used for rapid analysis of serum beta-glucans but also provided a screening procedure for the determination of fungal infections.
Liu
Ding
D
Department of Laboratory Medicine, Daping Hospital, Third Military Medical University, Chongqing, People's Republic of China. liudingcq@sohu.com <liudingcq@sohu.com>
Luo
Peng
P
Sun
Wenwen
W
Zhang
Liping
L
Wang
Zheng
Z
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D048271:Chitosan / Q000737:chemistry; D055664:Electrochemical Techniques / Q000379:methods*; D004566:Electrodes; D005465:Fluorocarbon Polymers / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D037181:Lectins, C-Type; D008565:Membrane Proteins / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry; D051379:Mice; D009181:Mycoses / Q000175:diagnosis; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D010640:Phenothiazines / Q000737:chemistry; D047071:beta-Glucans / Q000032:analysis* / Q000737:chemistry; D000099181:Dectin-1
D048271:Chitosan; D005465:Fluorocarbon Polymers; D006046:Gold; D037181:Lectins, C-Type; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D010640:Phenothiazines; D047071:beta-Glucans; D000099181:Dectin-1; C040402:perfluorosulfonic acid; C009469:thionine