A sensitive immunosorbent bio-barcode assay combining PCR with icELISA for detection of gonyautoxin 2/3.
In the current study, we developed a nanosphere bio-barcode technology to detect trace gonyautoxin 2/3 (GTX 2/3). GTX 2/3-glucose oxidase (GOX) conjugates were first prepared as the coating antigen in a periodate reaction. Subsequently, gold nanoparticles (NP) dual-labeled with anti-GTX 2/3 monoclonal antibodies (Mab) and DNA oligonucleotides were synthesized via a one-step preparation method. Combining PCR with indirect competitive ELISA (icELISA), a novel immunosorbent bio-barcode assay was established utilizing the Mab-NP-dsDNA complex to convert enzymatic signals to DNA signals. Importantly, the limit of detection of the method was lower than 0.74 microg mL(-1). Thus, the immunosorbent bio-barcode assay is a rapid and high-throughput screening tool to detect GTX 2/3 in aquatic products.
Tang
Yong
Y
Center of Antibody Engineering, Department of Bioengineering, Jinan University, Guangzhou 510632, PR China.
Wang
Hong
H
Xiang
Junjian
J
Chen
Yaoqiang
Y
He
Wenfei
W
Deng
Ning
N
Yang
Hongyu
H
D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D004247:DNA / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005949:Glucose Oxidase / Q000737:chemistry / Q000378:metabolism; D006046:Gold / Q000737:chemistry; D007164:Immunosorbents / Q000276:immunology / Q000378:metabolism; D053768:Metal Nanoparticles / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods*; D012530:Saxitoxin / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry
D000911:Antibodies, Monoclonal; D007164:Immunosorbents; D012530:Saxitoxin; D006046:Gold; D004247:DNA; D005949:Glucose Oxidase; C043405:gonyautoxin II