Dataset Information

Typing of Campylobacter jejuni isolated from poultry on the basis of flaA-RFLP by various restriction enzymes.

ABSTRACT: RFLP analysis of the flagellin (flaA) gene was compared using three different restriction endonucleases i.e DdeI, HinfI and DpnII to determine the genetic diversity among 43 Campylobacter jejuni isolates of poultry origin from the same geographical area. flaA gene was amplified in all the isolates and RFLP analysis showed variations. Dde-based RFLP was found most efficient in discriminating C. jejuni isolates by generating 15 different Dde-RFLP patterns with discriminatory index (D.I) of 0.9258 whereas DpnII produced seven Dpn-RFLP patterns (D.I .= 0.8427). While HinfI enzyme produced only six Hinf-RFLP patterns (D.I .= 0.6977). The discrimination of Dpn-RFLP was comparable to discrimination given by Dde-RFLP analysis, which is generally used to study flaA gene RFLP.

SUBMITTER: Yadav R

PROVIDER: S-EPMC7386692 | biostudies-literature | 2018 Dec

REPOSITORIES: biostudies-literature

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Typing of Campylobacter jejuni isolated from poultry on the basis of flaA-RFLP by various restriction enzymes.

Yadav Rahul R Yadav Jyotika J Maherchandani Sunil S Kashyap Sudhir Kumar SK

Veterinary and animal science 20180704

RFLP analysis of the flagellin (flaA) gene was compared using three different restriction endonucleases i.e DdeI, HinfI and DpnII to determine the genetic diversity among 43 Campylobacter jejuni isolates of poultry origin from the same geographical area. flaA gene was amplified in all the isolates and RFLP analysis showed variations. Dde-based RFLP was found most efficient in discriminating C. jejuni isolates by generating 15 different D ...[more]

PMID: 32734046

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Project description:We present an optimized multilocus sequence typing (MLST) scheme with universal primer sets for amplifying and sequencing the seven target genes of Campylobacter jejuni and Campylobacter coli. Typing was expanded by sequence determination of the genes flaA and flaB using optimized primer sets. This approach is compatible with the MLST and flaA schemes used in the PubMLST database and results in an additional typing method using the flaB gene sequence. An identification module based on the 16S rRNA and rpoB genes was included, as well as the genetic determination of macrolide and quinolone resistances based on mutations in the 23S rRNA and gyrA genes. Experimental procedures were simplified by multiplex PCR of the 13 target genes. This comprehensive approach was evaluated with C. jejuni and C. coli isolates collected in Switzerland. MLST of 329 strains resulted in 72 sequence types (STs) among the 186 C. jejuni strains and 39 STs for the 143 C. coli isolates. Fourteen (19%) of the C. jejuni and 20 (51%) of the C. coli STs had not been found previously. In total, 35% of the C. coli strains collected in Switzerland contained mutations conferring antibiotic resistance only to quinolone, 15% contained mutations conferring resistance only to macrolides, and 6% contained mutations conferring resistance to both classes of antibiotics. In C. jejuni, these values were 31% and 0% for quinolone and macrolide resistance, respectively. The rpoB sequence allowed phylogenetic differentiation between C. coli and C. jejuni, which was not possible by 16S rRNA gene analysis. An online Integrated Database Network System (SmartGene, Zug, Switzerland)-based platform for MLST data analysis specific to Campylobacter was implemented. This Web-based platform allowed automated allele and ST designation, as well as epidemiological analysis of data, thus streamlining and facilitating the analysis workflow. Data networking facilitates the exchange of information between collaborating centers. The described approach simplifies and improves the genotyping of Campylobacter, allowing cost- and time-efficient routine monitoring.

Dataset Information

Typing of Campylobacter jejuni isolated from poultry on the basis of flaA-RFLP by various restriction enzymes.

Publications

Typing of <i>Campylobacter jejuni</i> isolated from poultry on the basis of <i>fla</i>A-RFLP by various restriction enzymes.

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