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A combined polymerase chain reaction and restriction endonuclease enzyme assay for discriminating between Campylobacter coli and Campylobacter jejuni
Authors:Giuseppe Comi  Corrado Pipan  Giuseppe Botta  Luca Cocolin  Carlo Cantoni  Marisa Manzano
Affiliation:Dipartimento di Scienze degli Alimenti, Facoltàdi Agraria, Universitàdi Udine, Via Marangoni, 97, 33100 Udine, Italy;Cattedra di Microbiologia Medica, Facoltàdi Medicina, Universitàdi Udine, Udine, Italy;Istituto di Ispezione degli Alimenti di Origine Animale, Facoltàdi Medicina Veterinaria, Universitàdi Milano, Milan, Italy
Abstract:Abstract A combined polymerase chain reaction and restriction endonuclease (RE) enzyme assay was developed to discriminate between Campylobacter coli and Campylobacter jejuni . Amplimers of the FlaA gene obtained by PCR were digested with Alu I and Hin fI to distinguish C. coli from C. jejuni . With Alu I digestion C. jejuni -specific bands were observed at 110, 140 and 160 bp and C. coli -specific bands at 293 and 147 bp. C. jejuni -specific bands of 349 and 109 bp were found by Hin fI digestion but Hin fI did not digest the Fla A amplimer of C. coli . This combined technique is fast and easy to perform, and distinguishes the two campylobacters unequivocally.
Keywords:Campylobacter jejuni    Campylobacter coli    Polymerase chain reaction    Restriction endonuclease enzyme
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