Shen Z, Feng Y, Fox J G
Division of Comparative Medicine, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Helicobacter. 2000 Sep;5(3):121-8. doi: 10.1046/j.1523-5378.2000.00019.x.
Restriction fragment-length polymorphism (RFLP) analysis of a 1,200-bp polymerase chain reaction-amplified DNA fragment of gene coding for 16S rRNA was used to generate restriction profiles of 11 enterohepatic Helicobacter spp. isolated from various animals and humans.
The amplicon from each Helicobacter sp. was digested with four restriction endonucleases: Alu I, Hinf I, Hha I, and Dde I. Alu I digestion produced five patterns that were useful for initial differentiation.
Most Helicobacter spp. isolated from rodents had the same RFLP profiles by Alu I digestion (except H. rodentium and H. cholecystus), but they had different RFLP profiles by Hha I digestion. Only H. bilis and "H. rappini" mouse isolates could not be readily distinguished by the polymerase chain reaction-RFLP method. However, these two species can be distinguished using H. bilis specific primers. Some of the Helicobacter spp. have an intervening sequence in their 16S rRNA gene, which changes the RFLP patterns; in these cases, sequencing is the preferred method to make an appropriate diagnosis.
The RFLP method used in this study was straightforward and rapid and should prove useful as an adjunct for identification and classification of multiple enterohepatic Helicobacter spp.
对编码16S rRNA的基因进行1200bp聚合酶链反应扩增的DNA片段进行限制性片段长度多态性（RFLP）分析，以生成从各种动物和人类中分离出的11种肝螺杆菌属细菌的限制性图谱。
用四种限制性内切酶（Alu I、Hinf I、Hha I和Dde I）消化每种螺杆菌属细菌的扩增子。Alu I消化产生了五种有助于初步鉴别的模式。
通过Alu I消化，从啮齿动物中分离出的大多数螺杆菌属细菌具有相同的RFLP图谱（啮齿类螺杆菌和胆囊螺杆菌除外），但通过Hha I消化，它们具有不同的RFLP图谱。聚合酶链反应-RFLP方法无法轻易区分仅有的胆汁螺杆菌和“拉氏螺杆菌”小鼠分离株。然而，这两个菌种可以使用胆汁螺杆菌特异性引物进行区分。一些螺杆菌属细菌在其16S rRNA基因中有一个间隔序列，这会改变RFLP模式；在这些情况下，测序是进行准确诊断的首选方法。
本研究中使用的RFLP方法简单快速，应可作为多种肝螺杆菌属细菌鉴定和分类的辅助手段。
