title: Detection of Campylobacter species and Arcobacter butzleri in stool samples by use of real-time multiplex PCR.
abstract: The presence of Campylobacter (or Campylobacter-like) species in stools from patients suspected of infectious gastroenteritis (n = 493) was investigated using real-time PCR for detection of Arcobacter butzleri (hsp60 gene), Campylobacter coli (ceuE gene), Campylobacter jejuni (mapA), five acknowledged pathogenic Campylobacter spp. (C16S_Lund assay), and the Campylobacter genus (C16S_LvI assay). In total, 71.4% of the samples were positive for Campylobacter DNA (n = 352) by a Campylobacter genus-specific (C16S_LvI) assay. A total of 23 samples (4.7%) were positive in the C16S_Lund assay, used for detection of C. jejuni, C. coli, C. lari, C. upsaliensis, and C. hyointestinalis. Subsequent identification of these samples yielded detection frequencies (DF) of 4.1% (C. jejuni), 0.4% (C. coli), and 0.4% (C. upsaliensis). The DF of A. butzleri was 0.4%. Interestingly, sequencing of a subgroup (n = 46) of C16S_LvI PCR-positive samples resulted in a considerable number of Campylobacter concisus-positive samples (n = 20). PCR-positive findings with the C16S_Lund and C. jejuni/C. coli-specific assays were associated with more serious clinical symptoms (diarrhea and blood). Threshold cycle (C(T)) values of C. jejuni/C. coli PCR-positive samples were comparable to those of the C16S_Lund PCR (P = 0.21). C(T) values for both assays were significantly lower than those of the C16S_LvI assay (P < 0.001 and P < 0.00001, respectively). In conclusion, this study demonstrated that in combination, the C. jejuni/C coli-specific assays and the C16S_Lund assay are both useful for routine screening purposes. Furthermore, the DF of the emerging pathogen C. concisus was at least similar to the DF of C. jejuni.
authors.affiliation: Department of Research & Development, Laboratory for Infectious Diseases, Groningen, The Netherlands. r.de.boer@infectielab.nl
authors.forename: Richard F
authors.initials: RF
authors.lastname: de Boer
authors.forename: Alewijn
authors.initials: A
authors.lastname: Ott
authors.forename: Pinar
authors.initials: P
authors.lastname: Güren
authors.forename: Evert
authors.initials: E
authors.lastname: van Zanten
authors.forename: Alex
authors.initials: A
authors.lastname: van Belkum
authors.forename: Anna M D
authors.initials: AM
authors.lastname: Kooistra-Smid
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D020640:Arcobacter / Q000235:genetics / Q000302:isolation & purification*; D001431:Bacteriological Techniques / Q000379:methods*; D002167:Campylobacter / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D002648:Child; D002675:Child, Preschool; D005243:Feces / Q000382:microbiology*; D005260:Female; D005759:Gastroenteritis / Q000382:microbiology; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D008403:Mass Screening / Q000379:methods; D008875:Middle Aged; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D055815:Young Adult