Truett G E, Walker J A, Baker D G
Developmental Genetics Laboratory, Pennington Biomedical Research Center, Baton Rouge, LA 70808, USA.
Comp Med. 2000 Aug;50(4):444-51.
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.
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.
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.
These simple methods provide an efficient system for the development of Helicobacter-free mouse colonies.
为推动无幽门螺杆菌小鼠群落的发展，需要高效的方法来检测和消除幽门螺杆菌感染。我们开发了一种用于制备粪便DNA以进行幽门螺杆菌聚合酶链反应（PCR）检测的廉价、高通量方法。
通过在碱性溶液中将粪便颗粒加热至95摄氏度10分钟来制备粪便DNA，然后通过添加Tris缓冲液调节pH值。该溶液无需纯化DNA即可用于PCR检测。然后我们测试了寄养作为一种培育无幽门螺杆菌小鼠的方法。幽门螺杆菌阳性母鼠所生的幼崽在出生第一天就被转移到幽门螺杆菌阴性的寄养母鼠处。
寄养幼崽在89日龄前检测为幽门螺杆菌阴性，而由幽门螺杆菌阳性母鼠抚养的幼崽在19日龄时全部检测为阳性。
这些简单方法为建立无幽门螺杆菌小鼠群落提供了一个高效系统。
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