Comparative typing of Campylobacter jejuni by heat-stable serotyping and PCR-based restriction fragment length polymorphism analysis.
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.
Nakari
Ulla-Maija
UM
National Public Health Institute, Enteric Bacteria Laboratory, Mannerheimintie 166, FIN-00300 Helsinki, Finland.
Laaksonen
Katja
K
Korkeila
Maija
M
Siitonen
Anja
A
D015373:Bacterial Typing Techniques; D016123:Campylobacter jejuni / Q000145:classification* / Q000235:genetics; D006801:Humans; D008070:Lipopolysaccharides / Q000096:biosynthesis; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012703:Serotyping
D008070:Lipopolysaccharides; C023023:lipid-linked oligosaccharides