Mähler M, Bedigian H G, Burgett B L, Bates R J, Hogan M E, Sundberg J P
The Jackson Laboratory, Bar Harbor, Maine 04609-1500, USA.
Lab Anim Sci. 1998 Feb;48(1):85-91.
Four species of Helicobacter--H. muridarum, "H. rappini," H. hepaticus, and H. bilis--have been identified in the gastrointestinal tract of rodents. The association of Helicobacter species with chronic gastrointestinal diseases in mice has raised concern about their impact on research results. In this study, different methods for detection of Helicobacter species in the mouse intestinal tract were compared: polymerase chain reaction (PCR) amplification of 16S rRNA gene sequences, bacterial culture, electron microscopy, and histologic examination (Steiner stain). The PCR method was more sensitive in detecting murine Helicobacter species than was culture, electron microscopy, or histologic examination. Of the cecal specimens identified as Helicobacter species-positive by PCR, approximately 60% were identified as positive by each of the other methods. An 87.5% concordance was obtained by PCR screening of DNA from fecal and cecal specimens. Differentiation among murine Helicobacter species by colony morphologic or histologic features was not possible. Scanning electron microscopy and histologic examination indicated greater numbers of helical microorganisms, specifically H. hepaticus, in the cecum than in the colon. These results indicate that the PCR assays used can be performed on feces as a noninvasive means for rapidly screening large numbers of colony mice for murine Helicobacter species.
在啮齿动物的胃肠道中已鉴定出四种幽门螺杆菌——鼠幽门螺杆菌、“拉平尼幽门螺杆菌”、肝幽门螺杆菌和胆汁幽门螺杆菌。幽门螺杆菌与小鼠慢性胃肠道疾病的关联引发了人们对其对研究结果影响的担忧。在本研究中,比较了检测小鼠肠道中幽门螺杆菌的不同方法:16S rRNA基因序列的聚合酶链反应(PCR)扩增、细菌培养、电子显微镜检查和组织学检查(施泰纳染色)。PCR方法在检测鼠幽门螺杆菌方面比培养、电子显微镜检查或组织学检查更敏感。在通过PCR鉴定为幽门螺杆菌阳性的盲肠标本中,约60%通过其他每种方法鉴定为阳性。通过对粪便和盲肠标本的DNA进行PCR筛查,一致性达到87.5%。无法通过菌落形态或组织学特征区分鼠幽门螺杆菌种类。扫描电子显微镜检查和组织学检查表明,盲肠中的螺旋形微生物数量更多,特别是肝幽门螺杆菌,比结肠中的多。这些结果表明,所使用的PCR检测可在粪便上进行,作为一种非侵入性手段,用于快速筛查大量繁殖小鼠中的鼠幽门螺杆菌。