title: Specific detection of Vibrio parahaemolyticus by fluorescence quenching immunoassay based on quantum dots.
abstract: In this study, anti-Vibrio parahaemolyticus polyclonal and monoclonal antibodies were prepared through intradermal injection immune and lymphocyte hybridoma technique respectively. CdTe quantum dots (QDs) were synthesized at pH 9.3, 98 °C for 1 h with stabilizer of 2.7:1. The fluorescence intensity was 586.499, and the yield was 62.43%. QD probes were successfully prepared under the optimized conditions of pH 7.4, 37 °C for 1 h, 250 μL of 50 mg/mL EDC · HCl, 150 μL of 4 mg/mL NHS, buffer system of Na2HPO4-citric acid, and 8 μL of 2.48 mg/mL polyclonal antibodies. As gold nanoparticles could quench fluorescence of quantum dots, the concentration of V. parahaemolyticus could be detected through measuring the reduction of fluorescence intensity in immune sandwich reaction composed of quantum dot probe, gold-labeled antibody, and the sample. For pure culture, fluorescence intensity of the system was proportional with logarithm concentration of antigen, and the correlation coefficient was 99.764%. The fluorescence quenching immunoassay based on quantum dots is established for the first time to detect Vibrio parahaemolyticus. This method may be used as rapid testing procedure due to its high simplicity and sensitivity.
authors.affiliation: Key Laboratory of Environment Correlative Dietology of Ministry of Education, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Wang
authors.forename: Junxian
authors.initials: J
authors.lastname: Zhang
authors.forename: Haili
authors.initials: H
authors.lastname: Bai
authors.forename: Xuan
authors.initials: X
authors.lastname: Li
authors.forename: Pintian
authors.initials: P
authors.lastname: Lv
authors.forename: Ailing
authors.initials: A
authors.lastname: Guo
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D000918:Antibody Specificity; D019187:Cadmium Compounds / Q000737:chemistry; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000737:chemistry; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D007118:Immunoassay / Q000379:methods*; D053768:Metal Nanoparticles / Q000737:chemistry; D051379:Mice; D045663:Quantum Dots / Q000737:chemistry*; D011817:Rabbits; D013050:Spectrometry, Fluorescence; D013388:Succinimides / Q000737:chemistry; D013691:Tellurium / Q000737:chemistry; D013997:Time Factors; D014736:Vibrio parahaemolyticus / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000911:Antibodies, Monoclonal; D019187:Cadmium Compounds; D005456:Fluorescent Dyes; C465543:N-hydroxysulfosuccimide; D013388:Succinimides; D006046:Gold; D013691:Tellurium; D005022:Ethyldimethylaminopropyl Carbodiimide; C028337:cadmium telluride