Evaluation of a RAPD-based typing scheme in a molecular epidemiology study of Vibrio cholerae O1, Brazil

Leal, Nilma Cintra; Sobreira, Marise; Leal-Balbino, Tereza Cristina; Almeida, Alzira Maria Paiva de; Silva, M.J.B. da; Mello, Débora Medeiros; Seki, Liliane Miyuki; Hofer, Ernesto

doi:10.1111/j.1365-2672.2004.02090.x

Cited by 29 publications

(29 citation statements)

References 22 publications

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“…In previous study was observed that when the RAPD reactions are performed under rigorous reaction conditions (same preparation of genomic DNA, brand and lot of enzyme), the pattern of bands is reproducible 21 . In this study, standardized RAPD method showed to be reproductive and ascertained the diversity of the serogroups of the V. cholerae analyzed, which was demonstrated by the presence of different subtypes into the same serogroup.…”

Section: Discussionmentioning

confidence: 99%

Distribution of virulence markers in clinical and environmental Vibrio cholerae non-O1/non-O139 strains isolated in Brazil from 1991 to 2000

Theophilo

Rodrigues

Leal

et al. 2006

Rev. Inst. Med. trop. S. Paulo

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SUMMARYOne hundred seventy nine Vibrio cholerae non-O1/non-O139 strains from clinical and different environmental sources isolated in Brazil from 1991 to 2000 were serogrouped and screened for the presence of four different virulence factors. The Random Amplification of Polymorphic DNA (RAPD) technique was used to evaluate the genetic relatedness among strains. Fifty-four different serogroups were identified and V. cholerae O26 was the most common (7.8%). PCR analysis for three genes (ctxA, zot, ace) located of the CTX genetic element and one gene (tcpA) located on the VPI pathogenicity island showed that 27 strains harbored one or more of these genes. Eight (4.5%) strains possessed the complete set of CTX element genes and all but one of these belonged to the O26 serogroup suggesting that V. cholerae O26 has the potential to be an epidemic strain. The RAPD profiles revealed a wide variability among strains and no genetic correlation was observed.

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Section: Discussionmentioning

confidence: 99%

Distribution of virulence markers in clinical and environmental Vibrio cholerae non-O1/non-O139 strains isolated in Brazil from 1991 to 2000

Theophilo

Rodrigues

Leal

et al. 2006

Rev. Inst. Med. trop. S. Paulo

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“…Na pesquisa de genes associados à virulência de V. cholerae O 1 foi utilizada a metodologia preconizada por Keaster & Hall 5 com pequenas modificações 9 .…”

Section: Comunicação/communicationunclassified

Envolvimento de Aeromonas em surto de doença diarréica aguda em São Bento do Una, Pernambuco

Hofer

Reis

Theophilo

et al. 2006

Rev. Soc. Bras. Med. Trop.

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No primeiro semestre de 2004, ocorreu um surto de diarréia em São Bento do Una, Pernambuco, registrando-se 2.170 casos. Nas 582 coproculturas realizadas, 145 (25%) revelaram um enteropatógeno bacteriano, destacando 114 casos (19,5%) com a participação de Aeromonas, representadas por Aeromonas caviae (57/9,8%), Aeromonas veronii biovar sobria (23/3,9%), Aeromonas veronii biovar veronii (15/2,6%) e outras espécies (19/3,2%). Nos 31 episódios restantes (5,3%), foram detectados: V. cholerae O1 Ogawa toxigênico (18/3,1%), Salmonella spp (8/1,4%), Shigella spp (3/0,5%) e Vibrio cholerae não O1/não O139 (2/0,3%).

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“…Recent advances in biotechnology have resulted in the development of numerous methods for microorganism typing that differ in their sensitivity, rapidity, complexity, discriminatory power, reproducibility, labor-intensity, and cost (7,60,82,85). Several DNA-based methods were used for typing of V. cholerae, such as tests for virulence genes, ribotyping, pulsedfield gel electrophoresis (PFGE), amplified fragment length polymorphism, enterobacterial repetitive intergenic consensus sequence-PCR, random amplification of polymorphic DNA, and multilocus sequence typing (MLST) (2,17,34,44,46,67,73,87).…”

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confidence: 99%

Vibrio cholerae Strain Typing and Phylogeny Study Based on Simple Sequence Repeats

et al. 2007

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Vibrio cholerae is the etiological agent of cholera. Its natural reservoir is the aquatic environment. To date, practical typing of V. cholerae is mainly serological and requires about 200 antisera. Simple sequence repeats (SSR), also termed VNTR (for variable number of tandem repeats), provide a source of high genomic polymorphism used in bacterial typing. Here we describe an SSR-based typing method that combines the variation in highly mutable SSR loci, with that of shorter, relatively more stable mononucleotide repeat (MNR) loci, for accurate and rapid typing of V. cholerae. Vibrio cholerae, a gram-negative bacterium, is the causal agent of the severe diarrheal disease cholera. Its natural reservoir is the aquatic environment (49, 84). Cholera pandemics are caused by specific serogroups of V. cholerae that are pathogenic only to humans (35,84). Since 1817, V. cholerae has caused a number of pandemics. The first seven were presumed to be caused by O1 serogroup of V. cholerae (68). In October 1992, a new serogroup, defined O139, caused a severe outbreak of cholera in southeast India. Within 10 months, the O139 serogroup was disseminated all over the Indian subcontinent and soon thereafter spread to 11 neighboring countries, temporarily displacing the O1 serogroups (64). Since then both serogroups coexist and are responsible for large outbreaks. Genomic studies indicated that O139 epidemic strain arose by horizontal acquisition of unique DNA (15, 47).Traditionally, V. cholerae classification is serological and requires about 200 antisera based on the somatic O antigen (72). Isolates of V. cholerae are divided into three major subgroups: O1, O139, and non-O1/non-O139, of which only the O1 and O139 serogroups are associated with cholera pandemics and epidemics. Non-O1, non-O139 serogroups are recognized as causative agents of sporadic and localized outbreaks (73). Pathogenic V. cholerae isolates carry virulence genes, such as the toxins genes ctxAB (25,65,68). The environmental V. cholerae strains from non-O1, non-O139 serogroups are a possible natural reservoir of potentially new emerging epidemic strains (67,73). This assumption is supported by the finding that some of these environmental strains harbor virulence genes (23) and thus are likely to evolve into novel pathogenic strains by horizontal gene transfer (24,25). The emergence of new pathogenic V. cholerae strains requires not only an efficient, rapid, and accurate identification tool but also a means for determining genetic relationships among environmental and clinical isolates.Genome-based bacterial identification and typing is essential for several disciplines, including taxonomy, epidemiology, determining phylogenetic relationships, and the study of evolutionary mechanisms. It allows distinguishing among strains within a species to monitor epidemics and routes of contamination. Recent advances in biotechnology have resulted in the development of numerous methods for microorganism typing that differ in their sensitivity, rapidity, complexity, discrimin...

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Evaluation of a RAPD-based typing scheme in a molecular epidemiology study of Vibrio cholerae O1, Brazil

Cited by 29 publications

References 22 publications

Distribution of virulence markers in clinical and environmental Vibrio cholerae non-O1/non-O139 strains isolated in Brazil from 1991 to 2000

Distribution of virulence markers in clinical and environmental Vibrio cholerae non-O1/non-O139 strains isolated in Brazil from 1991 to 2000

Envolvimento de Aeromonas em surto de doença diarréica aguda em São Bento do Una, Pernambuco

Vibrio cholerae Strain Typing and Phylogeny Study Based on Simple Sequence Repeats

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