Comparative Typing of Campylobacter jejuni by Heat-Stable Serotyping and PCR-Based Restriction Fragment Length Polymorphism Analysis

Abstract

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.

Differences in Antimicrobial Resistance of Campylobacter Jejuni Isolated From Broiler Intestines and Drumsticks in Lithuania

“…This species also causes more campylobacteriosis cases in humans (Saenz et al . 2000; Nakari et al . 2005; Luangtongkum et al .…”

“…Campylobacter jejuni is the most prevalent species among campylobacters in poultry products (Van Looveren et al 2001). This species also causes more campylobacteriosis cases in humans (Saenz et al 2000;Nakari et al 2005;Luangtongkum et al 2007). Infected humans exhibit a range of clinical symptoms from mild, watery diarrhea to severe inflammatory diarrhea (Friedman et al 2004;Habib et al 2009).…”

A systematic review and meta-analysis of Penner serotype prevalence of Campylobacter jejuni in low- and middle-income countries

“…Further, the most readily available commercial typing kit for many laboratories, the Denka Seiken Campylobacter antisera kit, had the highest proportion of non-typable isolates of the methods reviewed here. The high proportion of non-typable strains may reflect isolates potentially typable by a broader screening array [ 42 , 94 ].…”

Application of molecular genetic methods in diagnostics and epidemiology of food‐borne bacterial pathogens

“…In our laboratory we have currently evaluated the applicability of a PCR‐RFLP‐method based on the lipo‐oligosaccharide (LOS) biosynthesis genes described by Shi et al (111). This method shows 100% typeability and correlation with Penner serotyping results, and therefore seems to be especially applicable in typing the strains not typeable by serotyping (112).…”