Molecular-based detection of non-culturable and emerging campylobacteria in patients presenting with gastroenteritis

Abstract

From January 2009 to May 2010, 436 faecal samples from patients with diarrhoeal illness in Southern Ireland were identified as Campylobacter genus-positive by an automated multiplex PCR; however, 204 (46·8%) of these samples were culture-negative for campylobacters. A combination of Campylobacter-specific uniplex PCR and 16S rRNA sequencing confirmed the presence of Campylobacter DNA in 191 (93·6%) of the culture-negative samples. Species-specific PCR identified C. jejuni (50·7%) C. ureolyticus (41%) and C. coli (5·7%) as the most prevalent species while C. fetus, C. upsaliensis, C. hyointestinalis and C. lari accounted for 10% of culture-negative samples; mixed Campylobacter spp. were detected in 11% of samples. We conclude that non-culturable Campylobacter spp. are responsible for a considerable proportion of human enteritis and the true incidence of infection is likely to be significantly underestimated where conventional Campylobacter culture methods are used in isolation.

Detection of Campylobacter in Stool and Determination of Significance by Culture, Enzyme Immunoassay, and PCR in Developing Countries

“…The majority of studies of Campylobacter infection have used selective culture techniques designed to improve isolation of Campylobacter jejuni and C. coli, which are thought to be the primary species associated with human disease (2). Recently, several antigen-based tests for the detection of Campylobacter have been developed, and in a European setting these were revealed to have excellent sensitivity and specificity compared with the results of culture (Ͼ89%) (3).…”

“…Though this is thought to be primarily a product of both false-positive enzyme immunoassay (EIA) results and the low sensitivity of Campylobacter culture, there is some evidence that increased detection of Campylobacter by antigen-based tests can represent detection of non-jejuni/coli Campylobacter species, specifically, C. upsaliensis (7). Finally, several studies have now used PCR tests to detect a diverse range of Campylobacter species of unclear pathogenicity in patients with and without gastroenteritis (2,8,9).…”

Campylobacter hyointestinalis Isolated from Pigs Produces Multiple Variants of Biologically Active Cytolethal Distending Toxin

“…In recent years, increasing numbers of C. hyointestinalis organisms have been isolated from various animal sources, including pigs, and from diarrheal patients, as well as from immunocompromised hosts (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13). However, to date no information is available about virulence factors and virulence mechanisms of C. hyointestinalis isolated from animals.…”

“…Because antimicrobial agents such as cephem antibiotics often are used in selective media, some campylobacters are difficult to isolate by currently avail-able methods. When PCR-based detection methods as well as nonselective media and hydrogen were employed, increasing numbers of C. hyointestinalis organisms were isolated from diarrheal patients, in particular from children and immunocompromised hosts (4,(7)(8)(9)(10)(11)(12). For this reason, the development of rapid detection methods for clinically important campylobacters, including C. hyointestinalis, is required (22,27).…”

“…Subsequently, this organism was isolated from not only diseased but also healthy animals (3)(4)(5)(6). Several recent reports indicate the isolation of increasing numbers of C. hyointestinalis organisms from patients with diarrhea (4,(7)(8)(9)(10)(11)(12). Therefore, C. hyointestinalis is considered an emerging zoonotic pathogen of the genus Campylobacter.…”