Evaluation of molecular assays for identification Campylobacter fetus species and subspecies and development of a C. fetus specific real-time PCR assay

Autor: van der Graaf-van Bloois, L., van Bergen, M.A.P., van der Wal, F.J., de Boer, A.G., Duim, B., Schmidt, T., Wagenaar, J.A., Advances in Veterinary Medicine, Strategic Infection Biology, Dep Infectieziekten Immunologie
Přispěvatelé: Advances in Veterinary Medicine, Strategic Infection Biology, Dep Infectieziekten Immunologie
Jazyk: angličtina
Rok vydání: 2013
Předmět:
Microbiology (medical)
AFLP
venerealis
Coronacrisis-Taverne
Cattle Diseases
polymerase-chain-reaction
Subspecies
Real-Time Polymerase Chain Reaction
Microbiology
Sensitivity and Specificity
law.invention
strains
Campylobacter fetus
law
Campylobacter Infections
Animals
Bovine genital campylobacteriosis
samples
Insertion sequence
genes
Molecular Biology
Polymerase chain reaction
reproductive and urinary physiology
Genetics
Host Pathogen Interaction & Diagnostics
Fetus
Bacteriological Techniques
biology
16s ribosomal-rna
Bacteriologie
Bacteriology
Bacteriology
Host Pathogen Interaction & Diagnostics
differentiation
biology.organism_classification
Subspecies identification
amplified fragment
Host Pathogen Interactie & Diagnostiek
jejuni
Real-time polymerase chain reaction
coli
Molecular Diagnostic Techniques
Bacteriologie
Host Pathogen Interactie & Diagnostiek
embryonic structures
Multilocus sequence typing
Amplified fragment length polymorphism
Cattle
MLST
Real-time PCR
Zdroj: Journal of Microbiological Methods 95 (2013) 1
Journal of Microbiological Methods, 95(1), 93-97
Journal of Microbiological Methods, 95, 93. Elsevier
ISSN: 0167-7012
Popis: Phenotypic differentiation between Campylobacter fetus (C. fetus) subspecies fetus and C. fetus subspecies venerealis is hampered by poor reliability and reproducibility of biochemical assays. AFLP (amplified fragment length polymorphism) and MLST (multilocus sequence typing) are the molecular standards for C. fetus subspecies identification, but these methods are laborious and expensive. Several PCR assays for C. fetus subspecies identification have been described, but a reliable comparison of these assays is lacking. The aim of this study was to evaluate the most practical and routinely implementable published PCR assays designed for C. fetus species and subspecies identification. The sensitivity and specificity of the assays were calculated by using an extensively characterized and diverse collection of C. fetus strains. AFLP and MLST identification were used as reference. Two PCR assays were able to identify C. fetus strains correctly at species level. The C. fetus species identification target, gene nahE, of one PCR assay was used to develop a real-time PCR assay with 100% sensitivity and 100% specificity, but the development of a subspecies venerealis specific real-time PCR (ISCfe1) failed due to sequence variation of the target insertion sequence and prevalence in other Campylobacter species. None of the published PCR assays was able to identify C. fetus strains correctly at subspecies level. Molecular analysis by AFLP or MLST is still recommended to identify C. fetus isolates at subspecies level.
Databáze: OpenAIRE
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