Evaluation of a PCR assay for identification and differentiation of <i>Campylobacter fetus</i> subspecies

Hum, S.; Quinn, K.; Brunner, J.; On, Stephen L. W.

doi:10.1111/j.1751-0813.1997.tb15665.x

Cited by 128 publications

(220 citation statements)

References 23 publications

(42 reference statements)

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“…Primers for species-level (forward MG3F, GGT AGC CGC AGC TGC TAA GAT; reverse MG4R, TAG CTA CAA TAA CGA CAA CT) and for subsp. venerealis (forward VenSF, CTT AGC AGT TTG CGA TAT TGC CAT T; reverse VenSR, GCT TTT GAG ATA ACA ATA AGA GCT T) identi®cation were used in a multiplex PCR reaction comprising 2 ll template (heat-lysed bacterial cells) and 48 ll PCR mastermix (®nal concentration 1´Perkin-Elmer PCR buffer; 0á1 mmol l )1 each dNTP; 3 mmol l )1 MgCl 2 ; 0á5 U Taq polymerase; 0á5 lmol l )1 each primer PFGE±DNA pro®ling and cluster analysis of patterns PFGE±DNA-containing blocks were prepared, digested with enzyme SmaI and the resulting genomic fragments separated by electrophoresis and visualized using methods described previously (Hum et al 1997). Photographs of ethidium bromide-stained PFGE±DNA pro®les were digitized using a desktop scanner (Deskjet II, Hewlett-Packard, CA, USA).…”

Section: Pcrmentioning

confidence: 99%

Evaluation of numerical analysis of PFGE-DNA profiles for differentiating Campylobacter fetus subspecies by comparison with phenotypic, PCR and 16S rDNA sequencing methods

On¹,

Harrington²

2001

J Appl Microbiol

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Aims: To assess the ef®cacy of numerical analysis of PFGE-DNA pro®les for identi®cation and differentiation of Campylobacter fetus subspecies. Methods and Results: 31 Camp. fetus strains were examined by phenotypic, PCR-and PFGE-based methods, and the 16S rDNA sequences of 18 strains compared. Numerical analysis of PFGE-DNA pro®les divided strains into two clusters at the 86% similarity level. One cluster contained 19 strains clearly identi®ed as Camp. fetus subsp. venerealis. The other cluster comprised 12 strains, of which 10 were unambiguously identi®ed as Camp. fetus subsp. fetus. The remaining two strains were identi®ed as Camp. fetus subsp. venerealis by either phenotypic or PCR methods, but not both. At higher similarity levels, clusters containing isolates from each of two countries were identi®ed, suggesting that certain clones predominate in certain geographical regions. Conclusions: Numerical analysis of PFGE-DNA pro®les is an effective method for differentiating Camp. fetus subspecies. Signi®cance and Impact of the Study: Critical comparison of PFGE, PCR, 16S rDNA sequencing and phenotypic methods for differentiation of Camp. fetus subspecies was attained. Novel phenotypic markers for distinguishing subspecies were identi®ed. Evidence for dominant clones of each subspecies in certain countries was provided.

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Section: Pcrmentioning

confidence: 99%

Evaluation of numerical analysis of PFGE-DNA profiles for differentiating Campylobacter fetus subspecies by comparison with phenotypic, PCR and 16S rDNA sequencing methods

On¹,

Harrington²

2001

J Appl Microbiol

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“…venerealis poses a special problem for veterinary laboratories. Although several phenotypic and genotypic methods are useful for discriminating these two subspecies (Hum et al, 1997;On & Harrington, 2001), the final determination is based mainly on the different pathogenic associations of the subspecies. C. fetus subsp.…”

mentioning

confidence: 99%

Genetic relatedness within the genus Campylobacter inferred from rpoB sequences

Korczak

Stieber

Emler

et al. 2006

International Journal of Systematic and Evolutionary Microbiology

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The genus Campylobacter comprises 17 species, some of which are important animal and human pathogens. To gain more insight into the genetic relatedness of this genus and to improve the molecular tools available for diagnosis, a universal sequencing approach was established for the gene encoding the beta-subunit of RNA polymerase (rpoB) for the genus Campylobacter. A total of 59 strains, including the type strains of currently recognized species as well as field isolates, were investigated in the study. A primer set specific for Campylobacter species enabled straightforward amplification and sequencing of a 530 bp fragment of the rpoB gene. The 16S rRNA gene sequences of all of the strains were determined in parallel. A good congruence was obtained between 16S rRNA and rpoB gene sequence-based trees within the genus Campylobacter. The branching of the rpoB tree was similar to that of the 16S rRNA gene tree, even though a few discrepancies were observed for certain species. The resolution of the rpoB gene within the genus Campylobacter was generally much higher than that of the 16S rRNA gene sequence, resulting in a clear separation of most species and even some subspecies. The universally applicable amplification and sequencing approach for partial rpoB gene sequence determination provides a powerful tool for DNA sequence-based discrimination of Campylobacter species.The genus Campylobacter was first proposed by M. Sebald and M. Véron in 1963 and included only the type species Campylobacter fetus and Campylobacter bubulus, now known as Campylobacter sputorum (Sebald & Véron, 1963). These taxa had formerly been classified as Vibrio species. In 1973, M. Véron and R. Chatelain included several misclassified Vibrio in the distinct Campylobacter genus based on serological, biochemical and DNA-DNA hybridization investigations (On, 2001). Since then, the taxonomy of the genus has changed dramatically. At present, it comprises 17 species with validly published names and six recognized subspecies (On, 2001;Foster et al., 2004). In addition, strains belonging to C. sputorum are divided into three biovars, sputorum, fecalis and paraureolyticus, on the basis of their ability to produce catalase or urease Vandamme & On, 2001).In general, members of the genus Campylobacter colonize the mucosal surfaces of the intestinal tract, oral cavity or urogenital tract of healthy, as well as diseased, humans and animals, especially birds. Several species may act as pathogens, causing disease in both human and animal hosts. Twelve of the 17 Campylobacter species are associated with human diseases. Campylobacter jejuni and Campylobacter coli are particularly frequent causative agents of human bacterial intestinal disorders worldwide (Skirrow, 1994). There have also been reported cases of diarrhoea in man caused by Campylobacter upsaliensis and Campylobacter lari, but the frequency of these infections is very low (Bourke et al., 1998;Van Doorn et al., 1998). Occasionally, Campylobacter species are implicated as causative agents of pericard...

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“…fetus a produção de H 2 S em meio contendo cistina e a tolerância a 1% de glicina, testes nos quais as subespécies são negativas e positivas paras ambos, respectivamente (Vandamme 2000). Além disto, a imunofluorescência direta (IFD) (Figueiredo et al 2002) e a reação em cadeia da polimerase (PCR) também podem ser utilizadas (Hum et al 1997.…”

Section: Diagnóstico Clínico E Laboratorialunclassified

“…fetus do C. fetus subsp. venerealis (Hum et al 1997) e PCR em tempo real com sondas específicas, também capazes de diferenciar as duas subespécies . Para a detecção de T. foetus, o material coletado deve ser inoculado e transportado em meio de transporte e enriquecimento de Diamond modificado, enriquecido com 10% de soro fetal bovino e mescla de antibióticos (penicilina e estreptomicina e anfotericina B) (Appel et al 1993).…”

Section: Diagnóstico Clínico E Laboratorialunclassified

Campilobacteriose genital bovina e tricomonose genital bovina: epidemiologia, diagnóstico e controle

et al. 2011

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Pesq. Vet. Bras. 31(4): 336-344, abril 2011 RESUMO.-A presente atualização trata de duas das mais importantes doenças sexualmente transmitidas de bovinos, a campilobacteriose genital bovina e a tricomonose genital bovina. São abordados aspectos relacionados à epidemiologia destas doenças, principalmente em relação a sua distribuição no Brasil. Também são revisados aspectos importantes de diagnóstico, incluindo as técnicas e interpretação dos resultados, além de medidas de controle para ambas as doenças. INTRODUÇÃOA campilobacteriose genital bovina (CGB) e a tricomonose genital bovina (TGB) são doenças infecto-contagiosas de transmissão sexual, que acometem bovinos em idade reprodutiva e causam grandes perdas econômicas em decorrência dos problemas reprodutivos desencadeados (Dekeyser 1984, Goodger & Skirrow 1986 The present update deals with two of the most important sexually transmitted diseases of cattle: bovine genital campylobacteriosis and bovine genital trichomonosis. Epidemiological aspects, mainly their distribution in Brazil, alongside with their diagnosis in cattle are presented and commented. The main points in their diagnoses, including the description of the techniques and the interpretation of the results are also reviewed. Finally the control and prevention of both diseases are discussed.

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Evaluation of a PCR assay for identification and differentiation of Campylobacter fetus subspecies

Cited by 128 publications

References 23 publications

Evaluation of numerical analysis of PFGE-DNA profiles for differentiating Campylobacter fetus subspecies by comparison with phenotypic, PCR and 16S rDNA sequencing methods

Evaluation of numerical analysis of PFGE-DNA profiles for differentiating Campylobacter fetus subspecies by comparison with phenotypic, PCR and 16S rDNA sequencing methods

Genetic relatedness within the genus Campylobacter inferred from rpoB sequences

Campilobacteriose genital bovina e tricomonose genital bovina: epidemiologia, diagnóstico e controle

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