Identifying IgA-coated Gut Bacteria

We used a validated method for detecting IgA-coated bacteria (12 (link)). Briefly, Mpbio Fastprep Lysing Matrix D Tubes (MP Biomedicals, Santa Ana, CA) were used to homogenize the feces, and anti-IgA-phycoerythrin (1:5; clone IS11-8E10; Miltenyi Biotec, Bergisch Gladbach, Germany) was used to stain the samples before flow cytometry (ACEA NovoCyte, San Diego, CA). Anti-phycoerythrin MicroBeads (Miltenyi Biotec) and a Quadro magnetic activated cell sorting (MACS) Starting Kit (LS; Miltenyi Biotec) were used to sort fecal bacteria. After MACS separation, the negative and positive portions were assembled for 16S rRNA gene sequencing analysis.

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Liu Y., Yuan X., Li L., Lin L., Zuo X., Cong Y, & Li Y. (2020). Increased Ileal Immunoglobulin A Production and Immunoglobulin A-Coated Bacteria in Diarrhea-Predominant Irritable Bowel Syndrome. Clinical and Translational Gastroenterology, 11(3), e00146.

Publication 2020

Anti-phycoerythrin MicroBeads

16s rrna Anti iga Bacteria Cell separation Clone Fecal Flow cytometry Gene Microbeads Phycoerythrin Sequencing analysis Stain

Corresponding Organization :

Other organizations : Qilu Hospital of Shandong University, Qingdao University, Affiliated Hospital of Qingdao University, The University of Texas Medical Branch at Galveston

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Variable analysis

independent variables

Homogenization method (Mpbio Fastprep Lysing Matrix D Tubes)
Staining method (anti-IgA-phycoerythrin)
Cell sorting method (Anti-phycoerythrin MicroBeads and Quadro magnetic activated cell sorting (MACS) Starting Kit)

dependent variables

Presence and quantification of IgA-coated bacteria
16S rRNA gene sequencing analysis of the sorted fecal bacteria

control variables

Concentration of anti-IgA-phycoerythrin (1:5)

positive controls

None specified

negative controls

None specified

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