Development of a colloidal gold immunochromatographic strip for rapid detection of Streptococcus agalactiae in tilapia

无乳链球菌 微生物学 猪链球菌 胶体金 生物 化学 嗜水气单胞菌 罗非鱼 细菌 链球菌 材料科学 纳米技术 遗传学 纳米颗粒 渔业
作者
Wende Wu,Min Li,Ming Chen,Liping Li,Rui Wang,Hailan Chen,Fuyan Chen,Qiang Mi,Liang Wan-wen,Hanzhong Chen
出处
期刊:Biosensors and Bioelectronics [Elsevier]
卷期号:91: 66-69 被引量:54
标识
DOI:10.1016/j.bios.2016.11.038
摘要

A colloidal gold immunochromatographic strip was developed for rapid detection of Streptococcus agalactiae (S. agalactiae) infection in tilapia. The monoclonal antibodies (mAb) 4C12 and 3A9 were used to target S. agalactiae as colloidal gold-mAb conjugate and captured antibody, respectively. The colloidal gold immunochromatographic strip was assembled via routine procedures. Optimal pH and minimum antibody levels in the reaction system for gold colloidal-mAb 4C12 conjugation were pH 7.4 and 18μg/mL, respectively. Optimal concentrations of the captured antibody 3A9 and goat anti-mouse antibody were 0.6mg/mL and 2mg/mL, respectively. The sensitivity of the strip for detecting S. agalactiae was 1.5×105 colony forming units (CFU). No cross-reaction was observed with other commonly encountered bacteria, including Pseudomonas fluorescens, Aeromonas hydrophila, Vibrio anguillarum and Streptococcus iniae. The assay time for S. agalactiae was less than 15min. Tilapia samples artificially infected with S. agalactiae were tested using the newly developed strip. The results indicated that blood, brain, kidney, spleen, metanephros and intestine specimens of infected fish can be used for S. agalactiae detection. The validity of the strip was maintained for 6 months at 4°C. These findings suggested that the immunochromatographic strip was effective for spot and rapid detection of S. agalactiae infected tilapia.

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