基于流式细胞术分选绵羊分泌型IgA包裹的消化道细菌

doi:10.11843/j.issn.0366-6964.2023.07.023

Abstract

Abstract: This study aimed to establish a flow cytometry (FCM) method for sorting secretory immunoglobulin A (sIgA) coated bacteria from the digestive tract contents of ruminants. Four healthy adult meat sheep in good physical condition were randomly selected and their ileal contents were collected to prepare the bacterial solution. The loading concentration of the bacterial solution, the working concentration of the bacterial dye 4', 6-diamidino-2-phenylindole (DAPI) and the anti-IgA antibody were optimized. The results showed that the optimal loading concentration of the bacterial solution of the contents was from 2.5 to 5 mg·mL^-1, the best staining concentration of DAPI was 5 μg·mL^-1, and the optimum amount of anti-IgA antibody was 5 μg. The reproducibility test results showed that the relative bias of the proposed method is less than 5% and that it is stable and reproducible. The sorting method of sIgA-coated bacteria established in this study is high selectivity, less sample consumption and easy operation and can provide technical support for related studies of the composition and function of sIgA-coated bacteria in ruminant digestive tracts.

Key words: secretory immunoglobulin A, secretory immunoglobulin A coated bacteria, microbe, flow cytometer

CLC Number:

S826.5

MI Hui, PENG Can, HE Zhixiong, TAN Zhiliang. Separation of Sheep Secretory Immunoglobulin A Coated Bacteria by Flow Cytometry[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 2924-2931.

References 34

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Separation of Sheep Secretory Immunoglobulin A Coated Bacteria by Flow Cytometry

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