Nakari Ulla-Maija, Laaksonen Katja, Korkeila Maija, Siitonen Anja
National Public Health Institute, Enteric Bacteria Laboratory, Mannerheimintie 166, FIN-00300 Helsinki, Finland.
J Clin Microbiol. 2005 Mar;43(3):1166-70. doi: 10.1128/JCM.43.3.1166-1170.2005.
Campylobacter jejuni has become the most common bacterial cause of human gastroenteritis worldwide. Rapid, discriminatory typing methods are required to identify potential clusters of infections. The major disadvantage of the well-evaluated and widely used Penner heat-stable serotyping method is the high level of nontypeability. The correlation of the types determined by the Penner heat-stable serotyping method and PCR-based restriction fragment length polymorphism (RFLP) analysis of the lipooligosaccharide (LOS) biosynthesis genes of C. jejuni was studied with 149 C. jejuni strains. Of these strains, 79 were patient strains belonging to 25 Penner serotypes, 60 were nontypeable patient strains, and 10 were reference strains. A 9.6-kb DNA fragment of the LOS gene cluster was amplified and digested with the restriction enzymes HhaI and DdeI. Altogether, 39 different RFLP types (including 30 HhaI profiles and 32 DdeI profiles) were identified. Type Hh1Dd1 was the most common type, with 36% of the strains and strains of 12 serotypes being of this type. A high level of discrimination was obtained, and a correlation between the Penner serotypes and the PCR-RFLP types could be seen. Also, variation in the LOS biosynthesis genes within a single Penner serotype was found. Although the PCR-RFLP method may not be sufficient to compensate for Penner serotyping, it can give valuable information about nontypeable strains and further characterize strains of common serotypes.
空肠弯曲菌已成为全球人类肠胃炎最常见的细菌病因。需要快速、具有鉴别力的分型方法来识别潜在的感染集群。经过充分评估且广泛使用的彭纳热稳定血清分型方法的主要缺点是不可分型率高。我们用149株空肠弯曲菌菌株研究了彭纳热稳定血清分型方法所确定的类型与基于聚合酶链反应(PCR)的空肠弯曲菌脂寡糖(LOS)生物合成基因限制性片段长度多态性(RFLP)分析所确定的类型之间的相关性。在这些菌株中,79株是属于25种彭纳血清型的患者菌株,60株是不可分型的患者菌株,10株是参考菌株。扩增了LOS基因簇的一个9.6 kb DNA片段,并用限制性内切酶HhaI和DdeI进行消化。总共鉴定出39种不同的RFLP类型(包括30种HhaI图谱和32种DdeI图谱)。Hh1Dd1型是最常见的类型,36%的菌株以及12种血清型的菌株属于该类型。获得了高度的鉴别力,并且可以看出彭纳血清型与PCR-RFLP类型之间存在相关性。此外,在单一彭纳血清型内发现了LOS生物合成基因的变异。虽然PCR-RFLP方法可能不足以弥补彭纳血清分型的不足,但它可以提供有关不可分型菌株的有价值信息,并进一步对常见血清型的菌株进行特征描述。