Quantification of Helicobacter pylori Load in an Infected Mouse Stomach

Published: September 29, 2023

Abstract

Source: D'Costa, K. et al., Mouse Models Of Helicobacter Infection And Gastric Pathologies. J. Vis. Exp. (2018)

In this video, we describe a protocol to quantify Helicobacter pylori load in mouse stomach tissues post-infection to confirm its colonization.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Confirmation of Bacterial Colonization in the Stomach Post-infection Viable Counting of H. pylori in the Stomach Supplement sterile HBA plates with additional antibiotics (200 μg/mL bacitracin and 10 μg/mL nalidixic acid) prior to performing colony counts from infected mou…

開示

The authors have nothing to disclose.

Materials

Bacteriological reagents
Oxoid Blood Agar Base No.2 Thermo Fischer Scientific CM0271B Dissolve in deinonized water prior to sterilization
Premium Defibrinated Horse blood Australian Ethical Biologicals PDHB100
Bacto Brain Heart Infusion Broth BD Bioscience 237500 Dissolve in deinonized water prior to sterilization
CampyGen gas packs Thermo Fischer Scientific CN0035A/CN0025A
Antibiotics
Vancomycin Sigma Aldrich V2002-1G Dissolve in deionized water
Polymyxin B Sigma Aldrich P4932-5MU Dissolve in deionized water
Trimethoprim (≥98% HPLC) Sigma Aldrich T7883 Dissolve in 100% (absolute) Ethanol
Amphotericin Amresco (Astral Scientific) E437-100MG Dissolve in deionized water
Bacitracin from Bacillus licheniformis Sigma Aldrich B0125 Dissolve in deionized water
Naladixic acid Sigma Aldrich N8878 Dissolve in deionized water
Equipment and plasticware
Oxoid Anaerobic Jars Thermo Fischer Scientific HP0011/HP0031
COPAN Pasteur Pipettes Interpath Services 200CS01
Eppendorf 5810R centrifuge Collect bacterial pellets by centrifugation at 2,200 rpm for 10 mins at 4°C
Parafilm M Bemis, VWR PM996
Eppendorf micropestle for 1.2 – 2 mL tubes Sigma Aldrich Z317314 Autoclavable polypropylene pestles used for stomach homogenization
GentleMACs Dissociator Miltenyi Biotec 130-093-235 Use a pre-set gentleMACS Programs for mouse stomach tissue
M Tubes (orange cap) Miltenyi Biotec 30-093-236
Qubit Fluorometer Thermo Fischer Scientific Q33216
Sterile plastic loop LabServ LBSLP7202

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記事を引用
Quantification of Helicobacter pylori Load in an Infected Mouse Stomach. J. Vis. Exp. (Pending Publication), e21711, doi: (2023).

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