L Petersen1, D G Newell. 1. Danish Veterinary Laboratory, Department of Poultry, Fish and Fur Animals, Aarhus, Denmark. lpe@svs.dk
Abstract
AIMS: The aim of this investigation was to compare the usefulness of two previously published flagellin PCR-RFLP typing (Fla-typing) techniques for the subtyping of Campylobacter jejuni strains, in terms of ease of use and discriminatory power. METHODS AND RESULTS: Six groups of isolates, which were epidemiologically unrelated but with similar Fla-types, and five groups of epidemiologically related poultry isolates, with similar PFGE profiles, were used in the comparison. The Fla-typing methods used varied in the number and length of fla-genes amplified and the restriction enzymes used. In addition, the use of separately amplified PCR fragments of both the flaA and flaB genes to generate RFLP profiles was investigated. CONCLUSION: The results clearly demonstrated that both previously published methods exhibit some advantages over the other. However, optimal discrimination was obtained by the use of separately amplified PCR fragments of both fla-genes. SIGNIFICANCE AND IMPACT OF THE STUDY: The subtyping of Camp. jejuni isolates is considered essential for epidemiological purposes. Genotyping methods are now more frequently used but have yet to be standardized. Fla-typing is a rapid and easy to use method with acceptable discriminatory power. However, the discriminatory power of the currently published Fla-typing techniques may be further improved by incorporating RFLP profiles of both fla-genes.
AIMS: The aim of this investigation was to compare the usefulness of two previously published flagellin PCR-RFLP typing (Fla-typing) techniques for the subtyping of Campylobacter jejuni strains, in terms of ease of use and discriminatory power. METHODS AND RESULTS: Six groups of isolates, which were epidemiologically unrelated but with similar Fla-types, and five groups of epidemiologically related poultry isolates, with similar PFGE profiles, were used in the comparison. The Fla-typing methods used varied in the number and length of fla-genes amplified and the restriction enzymes used. In addition, the use of separately amplified PCR fragments of both the flaA and flaB genes to generate RFLP profiles was investigated. CONCLUSION: The results clearly demonstrated that both previously published methods exhibit some advantages over the other. However, optimal discrimination was obtained by the use of separately amplified PCR fragments of both fla-genes. SIGNIFICANCE AND IMPACT OF THE STUDY: The subtyping of Camp. jejuni isolates is considered essential for epidemiological purposes. Genotyping methods are now more frequently used but have yet to be standardized. Fla-typing is a rapid and easy to use method with acceptable discriminatory power. However, the discriminatory power of the currently published Fla-typing techniques may be further improved by incorporating RFLP profiles of both fla-genes.
Authors: Clifford G Clark; Louis Bryden; Wilfred R Cuff; Patricia L Johnson; Frances Jamieson; Bruce Ciebin; Gehua Wang Journal: J Clin Microbiol Date: 2005-05 Impact factor: 5.948
Authors: Petra Griekspoor; Jenny Olofsson; Diana Axelsson-Olsson; Jonas Waldenström; Björn Olsen Journal: Appl Environ Microbiol Date: 2013-02-01 Impact factor: 4.792
Authors: Karen T Elvers; Christopher R Helps; Trudy M Wassenaar; Vivien M Allen; Diane G Newell Journal: Appl Environ Microbiol Date: 2008-02-15 Impact factor: 4.792
Authors: A J H Leatherbarrow; C A Hart; R Kemp; N J Williams; A Ridley; M Sharma; P J Diggle; E J Wright; J Sutherst; N P French Journal: Appl Environ Microbiol Date: 2004-02 Impact factor: 4.792
Authors: Hosny El-Adawy; Helmut Hotzel; Herbert Tomaso; Heinrich Neubauer; Eduardo N Taboada; Ralf Ehricht; Hafez M Hafez Journal: PLoS One Date: 2013-02-20 Impact factor: 3.240