Cardarelli-Leite P, Blom K, Patton C M, Nicholson M A, Steigerwalt A G, Hunter S B, Brenner D J, Barrett T J, Swaminathan B
Foodborne and Diarrheal Diseases Branch, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.
J Clin Microbiol. 1996 Jan;34(1):62-7. doi: 10.1128/jcm.34.1.62-67.1996.
Restriction fragment length polymorphism analysis of a PCR-amplified DNA fragment of the gene coding for 16S rRNA was performed on 148 previously characterized strains of Campylobacter, Helicobacter, Arcobacter, and Wolinella succinogenes and 13 Campylobacter-like isolates. These strains included clinical, animal, and environmental isolates. PCR amplification generated a 283-bp fragment from all species. The amplicon from each strain was digested with six restriction endonucleases (AccI, AvaI, DdeI, HaeIII, HpaII, XhoI). DdeI was useful for the initial grouping of the strains. Additional discrimination within the different DdeI groups was obtained with AccI, HaeIII, HpaII, and XhoI digestions. The PCR-restriction fragment length polymorphism analysis allowed for the discrimination of members of the genus Campylobacter from members of closely related genera and discrimination between Campylobacter species. The proposed method is simple and rapid and can be useful for the routine identification of Campylobacter-like organisms in clinical or epidemiologic studies.
对编码16S rRNA的基因的PCR扩增DNA片段进行限制性片段长度多态性分析,该分析针对148株先前已鉴定的弯曲杆菌属、幽门螺杆菌属、弓形杆菌属和琥珀酸沃林氏菌菌株以及13株弯曲杆菌样分离株开展。这些菌株包括临床、动物和环境分离株。PCR扩增从所有物种中产生了一个283 bp的片段。用六种限制性内切酶(AccI、AvaI、DdeI、HaeIII、HpaII、XhoI)消化每个菌株的扩增子。DdeI有助于菌株的初步分组。通过AccI、HaeIII、HpaII和XhoI消化在不同的DdeI组内获得了进一步的区分。PCR-限制性片段长度多态性分析能够区分弯曲杆菌属成员与密切相关属的成员,并区分弯曲杆菌属的不同物种。所提出的方法简单快速,可用于临床或流行病学研究中弯曲杆菌样生物体的常规鉴定。