Application of PCR-RFLP provides as discrimination as totalflaA sequence analysis among C. jejuniisolates of clinical origin.

Application of PCR-RFLP provides as discrimination as totalflaA sequence analysis among C. jejuniisolates of clinical origin.
Jpn J Infect Dis. 2016 Jan 8;
Authors: Ghorbanalizadgan M, Bakhshi B, Najar-Peerayeh S
Abstract
The aim of this study was to determinethegenetic relatedness amongtwenty clinical Campylobacterjejuniisolated from children with diarrhea in Iran,and to introduce the best discriminatory method based on flaA sequence divergence.A total of 400 stool specimens were obtained from children under five years of age during July 2012 to June 2013. Primers were designedbased on conserved sequences flanking the flaA gene which encompass and amplify entire flaAgenefollowed bysequencing and analysis with MEGA version 6.0.6 software.Ninetyamino acid versus 560 nucleotidepolymorphic sequenceswere detectedwithin 1681bp of flaA sequence among which 43 (2.5%) and 12 (0.7%) were singleton.New repeat boxes within flaA sequences were found in this study.UPGMA dendrogram based on nucleotides of full length flaAgene,flaA SVR gene and in silicoflaA phylogenic tree of DdeI RFLP profilesproduced very similarclustering with diversity indexof 0.86 for each of three methods. It can be concluded that flaA typing based on DdeI RFLP of the PCR products, can be applied as cheap, rapid and reliable method for epidemiological study of C. jejuni isolates of clinical origin in resource limiting regions or in large scale population surveillances.
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Source: Japanese Journal of Infectious Diseases - January 8, 2016 Category: Infectious Diseases Authors: Ghorbanalizadgan M, Bakhshi B, Najar-Peerayeh S Tags: Jpn J Infect Dis Source Type: research