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Eradication of infection with Helicobacter spp. by use of neonatal transfer
G E Truett1, J A Walker, D G Baker
1Developmental Genetics Laboratory, Pennington Biomedical Research Center, Baton Rouge, LA 70808, USA.
Background And Purpose:
Efficient methods for detection and elimination of Helicobacter spp. infections are needed to facilitate the development of Helicobacter-free mouse colonies. We developed an inexpensive, high-throughput method for preparation of fecal DNA for Helicobacter polymerase chain reaction (PCR) assays.
Methods:
Fecal DNA was prepared by heating fecal pellets to 95 degrees C for 10 minutes in an alkaline solution, then adjusting the pH by addition of Tris buffer. This solution is used for PCR assays without purification of DNA. We then tested fostering as a method of generating Helicobacter-free mice. Litters born to Helicobacter-positive dams were transferred to Helicobacter-negative foster dams on the first day of life.
Results:
Fostered pups tested Helicobacter negative up to 89 days of age, whereas pups raised by Helicobacter-positive dams were all test positive by 19 days of age.
Conclusion:
These simple methods provide an efficient system for the development of Helicobacter-free mouse colonies.
Insights
Developing Helicobacter-free mouse colonies is crucial. A new, inexpensive fecal DNA preparation method and fostering technique efficiently detect and eliminate Helicobacter spp. infections in mice.
Area of Science:
- Veterinary Microbiology
- Animal Health
- Molecular Diagnostics
Background:
- Helicobacter spp. infections pose a challenge for maintaining Helicobacter-free mouse colonies.
- Efficient detection and elimination methods are essential for research integrity.
Purpose of the Study:
- To develop a cost-effective, high-throughput method for fecal DNA preparation for Helicobacter polymerase chain reaction (PCR) assays.
- To establish an effective fostering strategy for generating Helicobacter-free mice.
Main Methods:
- A novel method involving heating fecal pellets in an alkaline solution followed by pH adjustment for DNA extraction was employed.
- PCR assays were performed directly on the prepared solution without DNA purification.
- A fostering program was implemented, transferring litters from infected dams to negative foster dams at birth.
Main Results:
- The developed fecal DNA preparation method proved efficient for Helicobacter PCR.
- Pups transferred to negative foster dams remained Helicobacter-negative up to 89 days of age.
- Pups raised by infected dams became positive by 19 days of age.
Conclusions:
- The combined methods offer a simple and efficient system for establishing Helicobacter-free mouse colonies.
- This approach supports the production of reliable animal models for research.