Genotyping of
            <i>Campylobacter</i>
            spp

Wassenaar, Trudy M.; Newell, Diane G.

doi:10.1128/aem.66.1.1-9.2000

Cited by 371 publications

(327 citation statements)

References 91 publications

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“…They also indicated a possible role for environmental reservoirs, such as water and flies, in the transmission of Campylobacter strains to poultry and other host sources (Hald et al, 2008;Newell & Fearnley, 2003). Difficulties in the reproducibility of methodology and interpretation of results among laboratories, however, precluded these being used as unified typing schemes by which Campylobacter isolates could be compared on a wider scale (Wassenaar & Newell, 2000).…”

Section: Introductionmentioning

confidence: 99%

Campylobacter sequence typing databases: applications and future prospects

Colles

Maiden

2012

Microbiology

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Section: Introductionmentioning

confidence: 99%

Campylobacter sequence typing databases: applications and future prospects

Colles

Maiden

2012

Microbiology

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“…The amplification of the flaA gene using consensus primers, and the digestion of the amplicon with DdeI were carried out as described by Wassenaar & Newell (2000). The digested products were run in a 2 % agarose gel at 1 V cm 21 for 120 min.…”

Section: Introductionmentioning

confidence: 99%

High level of ciprofloxacin resistance and its molecular background among Campylobacter jejuni strains isolated in the United Arab Emirates

Sonnevend

Rotimi

Kolodziejek

et al. 2006

Journal of Medical Microbiology

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The antibiotic sensitivity and the serotype and molecular type (MT) distribution of 41 Campylobacter jejuni strains isolated from individual patients in Tawam Hospital, Al Ain, United Arab Emirates, were investigated. While all strains were sensitive to erythromycin (MIC 0?5-4 mg l "1 ), 35 isolates (85?4 %) exhibited resistance to ciprofloxacin (MIC 8-64 mg l "1 ). All resistant strains carried the Thr-86 to Ile mutation in the gyrase A (gyrA) gene, as shown by mismatch amplification mutation assay (MAMA) and confirmed by sequencing. Based on the partial sequences of gyrA, resistant isolates carried 10 distinct alleles, eight of them representing new variants. Strains were assigned to 30 MTs based on the combined results of PFGE and flaA PCR-RFLP typing. Eight of the 35 ciprofloxacin-resistant strains, isolated over a period of more than 1 year, represented the largest MT, also carrying the same allelic variant of the gyrA gene. These results show that the local incidence of fluoroquinolone resistance among C. jejuni is one of the highest reported worldwide. It was also demonstrated that stable MTs could persist for a relatively long time among the clonally unrelated antibiotic-resistant isolates of C. jejuni. The data also emphasize the need to replace fluoroquinolones as empirical therapy for diarrhoea of undiagnosed aetiology. INTRODUCTIONFluoroquinolone resistance in Campylobacter jejuni, a leading cause of gastrointestinal infections worldwide and one of the most important causes of travellers' diarrhoea, is rapidly increasing. Resistance is commonly due to point mutations in the quinolone resistance determinant region (QRDR) of the gyrase A (gyrA) gene involving the Thr-86 for high, and the Asp-90 and Ala-70 amino acid residues for lower-level resistance (Engberg et al., 2001).While C. jejuni infections usually present as acute selflimiting gastrointestinal illnesses, in severe or prolonged cases antibiotic therapy is indicated, with macrolides as the primary drug of choice (Butzler, 2004). However, particularly if the causative agent is not identified, as is often the case in travellers' diarrhoea, fluoroquinolones are frequently used empirically (Guerrant et al., 2001;Yates, 2005). In such cases, resistance to ciprofloxacin can result in therapeutic failure (Guerrant et al., 2001;Sanders et al., 2002). Moreover, by mechanisms yet to be elucidated, fluoroquinolone resistance appears to coincide with increased fitness of the organism. In humans, fluoroquinolone-resistant strains are shed longer than sensitive isolates (Nelson et al., 2004;Engberg et al., 2004), and they have been reported to out-compete their sensitive counterparts, at least in certain genetic backgrounds, when colonizing poultry, i.e. the natural reservoir of the pathogen (Luo et al., 2005). All these findings underscore the necessity for close monitoring of the incidence and spread of fluoroquinolone-resistant clones of C. jejuni. Campylobacter isolates recovered from the 3389 stool samples tested during this period of time. Seven of th...

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“…Moreover, it is an expensive and laborious technique, demands a large number of standard strains and constant production of anti-serum. It also requires at least 5 to 7 days to be completed and repetitive subculture of the isolates before serological tests involving more than 74 different serotypes of C. jejuni may be performed (7,24). Table 2 shows that the 66 C. jejuni sequences that came from state of São Paulo were grouped in 44 subtypes identical by means of alignment and translation of nucleotide sequences.…”

Section: Discussionmentioning

confidence: 99%

Molecular subtyping of Campylobacter jejuni subsp. jejuni strains isolated from different animal species in the state of São Paulo, Brazil

Scarcelli¹,

Piatti²,

Harakava³

et al. 2005

Braz. J. Microbiol.

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The objective of the present trial was to characterize genetically strains of Campylobacter jejuni subsp. jejuni isolated from humans and several animal sources (bovines, swine, dogs, primates, wild boars and poultry). A total of 828 different animal samples (feces, carcass, aborted fetus and hysterectomized uterus) were analysed by means of routine bacteriological methods, and 36 C. jejuni strains were isolated. Thirty strains of human fecal origin were obtained in clinical analysis laboratories in the city of São Paulo. The 66 C. jejuni strains isolated were submitted to genetic characterization. Primers based on fla A gene were used in a polymerase chain reaction (PCR) and amplified a fragment of the 702 bp. PCR products were evaluated by means of sequencing and genealogic analysis. Genetic variability analysis of 66 strains showed 44 different subtypes of C. jejuni. One subtype was identical to a C. jejuni strain of human origin with the sequence in the GenBank (GENBANK -accession number AF050186). Subtyping analysis of C. jejuni strains based on sequencing of the fla A gene variable region and analysis of sequence alignment by the Maximum Parsimony method showed to be highly discriminatory, providing the best conditions to differentiate strains involved in outbreaks from those sporadically isolated. This is the first study of molecular subtyping analysis of human and animal C. jejuni strains using sequencing technique and genealogic analysis in the state of São Paulo, Brazil.

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Genotyping of Campylobacter spp

Cited by 371 publications

References 91 publications

Campylobacter sequence typing databases: applications and future prospects

Campylobacter sequence typing databases: applications and future prospects

High level of ciprofloxacin resistance and its molecular background among Campylobacter jejuni strains isolated in the United Arab Emirates

Molecular subtyping of Campylobacter jejuni subsp. jejuni strains isolated from different animal species in the state of São Paulo, Brazil

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