Multiplex PCR for detection of antibiotic resistance genes and the SXT element: application in the characterization of Vibrio cholerae

Ramachandran, Dhanya; Bhanumathi, R.; Singh, D. V.

doi:10.1099/jmm.0.46655-0

Cited by 44 publications

(37 citation statements)

References 22 publications

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“…The bacterial cell lysates were used as template DNA in all PCR assays. The presence of antibiotic resistance genes and SXT genetic element was determined by multiplex PCR (Ramachandran et al, 2007). The presence of virulence and regulatory genes ctxA, zot, ace, tcpA, ompU and toxR was determined by a hexaplex PCR assay using primers as previously published (Singh et al, 2002).…”

Section: Methodsmentioning

confidence: 99%

Phenotypic and genetic characteristics of Vibrio cholerae O1 carrying Haitian ctxB and attributes of classical and El Tor biotypes isolated from Silvassa, India

Das

Bhotra

Zala³

et al. 2016

Journal of Medical Microbiology

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Vibrio cholerae O1 biotype El Tor, the causative agent of the seventh pandemic, has recently been replaced by strains carrying classical and Haitian ctxB in India, Haiti and other parts of the world. We conducted phenotypic and genetic tests to characterize V. cholerae O1 isolated between 2012 and 2014 from Silvassa, India, to examine the presence of virulence and regulatory genes, seventh pandemic marker, ctxB type and biofilm formation and to study genomic diversity. Of the 59 V. cholerae O1, eight isolates belong to El Tor prototype, one to classical prototype and the remaining isolates have attributes of both classical and El Tor biotypes. PCR and ctxB gene sequencing revealed the presence of classical ctxB in four strains and Haitian ctxB in 55 isolates; indicating that isolates were either an El Tor or hybrid variant. All isolates carried virulence, regulatory, adherence, Vibrio seventh pandemic pathogenicity island I and seventh pandemic group-specific marker VC2346, in addition to tcpA ET and rstR ET , the features of seventh pandemic strains, and produced cholera toxin and biofilm. PFGE analysis showed that the majority of isolates are clonal and belong to fingerprint pattern A; however, pattern B is unrelated and patterns C and D are distinct, suggesting considerable diversity in the genomic content among them. These data thus show that isolates from Silvassa are genetically diverse and that Haitian ctxB and hybrid phenotypes are undergoing global dissemination.

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Section: Methodsmentioning

confidence: 99%

Phenotypic and genetic characteristics of Vibrio cholerae O1 carrying Haitian ctxB and attributes of classical and El Tor biotypes isolated from Silvassa, India

Das

Bhotra

Zala³

et al. 2016

Journal of Medical Microbiology

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“…However, the high genetic variability of microorganisms enables them to rapidly evade the action of antibiotics by developing antibiotic resistance [18]. One study revealed that all Vibrio strains were found to harbor antibiotics resistant genes and showed resistances to ampicillin, furazolidone, nalidixic acid, streptomycin, trimethoprim-sulfamethoxazole, and trimethoprim [19]. Thungapathra et al indicated that in a total number of 94 isolates of V. cholera, 43 strains contained R-plasmids and exhibited resistances to ampicillin, neomycin, tetracycline, gentamicin, streptomycin, sulfonamide, furazolidone, and chloramphenicol [20].…”

Section: Discussionmentioning

confidence: 99%

Antibiogram and Molecular Characterization of Vibrio Cholerae Isolated From Marine Fish

Angelo

Ramesh

2017

Asian J Pharm Clin Res

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Objective: Molecular identification and antibiotic susceptibility evaluation of Vibrio cholerae from marine fish available in local fish market Thanjavur, Tamil Nadu, India.Methods: Inoculation was done using nutrient agar as general media and thiosulfate-citrate-bile salts-sucrose agar as selective media and confirmed as V. cholerae by Gram-stain (microscopic observation), growth characteristics of different media, biochemical tests such as methyl red test, nitrate reduction tests, and indole test. Sensitivity (drug sensitivity) was done in Mueller-Hinton agar using disc diffusion method 10 different antibiotics were used to evaluate the antibiogram profile, molecular detection was done by targeting 16S rRNA gene using a universal primer.Results: V. cholerae is present in marine fish samples, as showed by culture method and microscopic observation as well biochemical tests. Polymerase chain reaction (PCR) amplification of 16S rRNA gene showed the amplification of targeted gene and antibiogram profile showed that isolates are more sensitive to ampicillin in comparison with others antibiotics used in this study. Ampicillin can be used for V. cholerae infection by the physicians and amoxicillin must be avoided which is resistant. Conclusion:Molecular detection is safe and rapid methods for bacteria identification as revealed by PCR amplification of 16S rRNA gene. As the isolates are more sensitive to ampicillin in comparison with others antibiotics used in this study. Ampicillin can be used for V. cholerae infection by the physicians and amoxicillin, and nitrofurantoin must be avoided.

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“…During the last 10 years the development of new antimicrobial drugs has been slowed down but the prevalence of resistance has been increased considerably. 4 Therefore, preventive actions must be taken to address this problem through developing new drugs, which may be either synthetic or natural. In this regard herbal medicines would be an alternate promising choice over synthetic drugs.…”

Section: Introductionmentioning

confidence: 99%

Leaf Extract of Syzygium cumini Shows Anti-Vibrio Activity Involving DNA Damage

Ahsan

Paul

Islam

et al. 2012

Dhaka Univ. J. Pharm. Sci

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ABSTRACT:The aim of the study was to investigate the effect of an ethanol extract of leaf (EEL) of Syzygium cumini against Vibrio cholerae serogroups Ogawa and Inaba. The antimicrobial activity of EEL was evaluated by the disc diffusion method against multi-drug resistant Ogawa and Inaba. The EEL effectively inhibited the growth of both serogroups. This growth inhibition was accompanied by fragmentation of genomic DNA as revealed by agarose gel electrophoresis. This result suggested that the EEL might inhibit bacterial growth involving DNA damage either through activation of signal transduction pathways or through direct interaction of the metabolites present in the EEL with DNA. Therefore, EEL of S. cumini has potential growth inhibitory activity against multi drug resistant Vibrios. This inhibitory effect of EEL might be explored to develop effective candidate(s) to combat cholera.

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Multiplex PCR for detection of antibiotic resistance genes and the SXT element: application in the characterization of Vibrio cholerae

Cited by 44 publications

References 22 publications

Phenotypic and genetic characteristics of Vibrio cholerae O1 carrying Haitian ctxB and attributes of classical and El Tor biotypes isolated from Silvassa, India

Phenotypic and genetic characteristics of Vibrio cholerae O1 carrying Haitian ctxB and attributes of classical and El Tor biotypes isolated from Silvassa, India

Antibiogram and Molecular Characterization of Vibrio Cholerae Isolated From Marine Fish

Leaf Extract of Syzygium cumini Shows Anti-Vibrio Activity Involving DNA Damage

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