地方株马驽巴贝斯虫Bc48截短体单克隆抗体的制备及应用

doi:10.11843/j.issn.0366-6964.2019.09.016

Abstract

Abstract: This study aimed to establish a rapid and accurate detection method for Babesia caballi. Six-week-old female BALB/c mice were immunized with purified Bc48 recombinant protein to prepare monoclonal antibodies, and a CI-ELISA method was established by Bc48 recombinant antigen and monoclonal antibody. The results showed that three hybridoma cell lines stably secreting monoclonal antibodies were prepared and named as 1H2, 7F4 and 11F4. By screening for CI-ELISA conditions, the optimal coating concentration of the antigen was 0.19 μg·mL^-1, and the optimal working concentration of monoclonal antibody 11F4 was 1:3.2×10⁵. The CI-ELISA was determined to have a critical value of 45% by detecting 30 Babesia caballi negative sera and 20 positive sera. Specificity test showed that the CI-ELISA does not reacted with positive sera from Theileria equi infected horses. Using the established CI-ELISA to detected 90 clinical sera, the total coincidence rate with the standard c-ELISA kit was 92.2%, the positive coincidence rate was 92.1%, and the negative coincidence rate was 94.1%. This results indicated that CI-ELISA method has characteristics of strong specificity, high sensitivity, good stability and repeatability, and simple operation. These results suggested that CI-ELISA established by monoclonal antibody (11F4) and recombinant protein (His-Bc48) was specific and reproducible, which could provide an effective means for the detection and monitoring of Babesia caballi infection in XinJiang, China.

CLC Number:

S852.723

WANG Panju, FAN Xinli, ZHANG Mengyuan, SONG Jingjing, LI Min, WU Lijiang, Bayin Chahan. Preparation and Application of Monoclonal Antibody against Truncated Bc48 of Babesia caballi Local Strains[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(9): 1882-1887.

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Preparation and Application of Monoclonal Antibody against Truncated Bc48 of Babesia caballi Local Strains

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