Development of a Hexaplex PCR Assay for Rapid Detection of Virulence and Regulatory Genes in Vibrio cholerae and Vibrio mimicus

  title={Development of a Hexaplex PCR Assay for Rapid Detection of Virulence and Regulatory Genes in Vibrio cholerae and Vibrio mimicus},
  author={D. V. Singh and Sree Renjini Isac and Rita R. Colwell},
  journal={Journal of Clinical Microbiology},
  pages={4321 - 4324}
ABSTRACT We have developed a hexaplex PCR assay for rapid detection of the virulence and regulatory genes for cholera toxin enzymatic subunit A (ctxA), zonula occludens toxin (zot), accessory cholera enterotoxin (ace), toxin-coregulated pilus (tcpA), outer membrane protein (ompU), and central regulatory protein ToxR (toxR) in Vibrio cholerae and Vibrio mimicus. This hexaplex PCR proved successful in screening pathogenic-toxigenic and nonpathogenic-nontoxigenic V. cholerae and V. mimicus strains… 

Quadruplex PCR for Simultaneous Detection of Serotype, Biotype, Toxigenic Potential, and Central Regulating Factor of Vibrio cholerae

A quadruplex PCR was developed for the simultaneous detection of genes specific for Vibrio cholerae O1 and/or O139 serogroup, cholera toxin A subunit, toxin-coregulated pilus, and central regulating protein ToxR (toxR) in a single tube reaction.

Rapid Detection of Virulence-Associated Genes in Environmental Strains of Vibrio cholerae by Multiplex PCR

A multiplex PCR assay was developed for the detection of virulence-associated genes (rtxA, tcpA, ctxA, hlyA, and sto) in environmental isolates of V. cholerae and showed that this method represents a simple, cost effective, and robust tool for rapid detection.

Septaplex PCR assay for rapid identification of Vibrio cholerae including detection of virulence and int SXT genes.

The one-step PCR described here can be used to identify V. cholerae accurately and rapidly, and the virulence and intsxt genes can be simultaneously detected, providing a useful method for monitoring pathogenic, intSxt-positive and nonpathogenic,intsxt-negative V. Cholerae serogroups both in the environment and clinical settings.

Analysis of lolB gene sequence and its use in the development of a PCR assay for the detection of Vibrio cholerae.

Detection of Virulence Genes in Vibrio cholerae Isolated from Aquatic Environment in Kerala, Southern India

The findings demonstrate that the virulence genes are dispersed among the environmental strains of V. cholerae and a complex aquatic environment can give rise to pathogenic V. Cholerae.

Rapid Screening of toxigenic and non-toxigenic V. cholera from clinical and environmental samples in Iraq by PCR

Zonnla occludens toxin (zot) is one of the major virulence factors of Vibrio cholera. The detection of zot producing V. cholerae using conventional culture, biochemical and immunological based assays

Characterization of a Toxigenic Vibrio cholerae O139 Strain Belonging to a New Ribotype and Isolated from a Diarrheal Patient

Molecular evidence is provided showing that toxigenic V. cholerae O139 strain ALO95 belongs to a distinct genotype characterized by a unique ribotype designated B-VII and has a unique enterobacterial repetitive intergenic consensus sequence PCR fingerprint profile designated E-V.

Detection of Vibrio cholerae by Real-Time Nucleic Acid Sequence-Based Amplification

Results indicate that NASBA can be used in the rapid detection of V. cholerae from various environmental water samples and has a strong potential for detecting toxigenic strains by using the tcpA and ctxA markers.

Design of a multiplex PCR method for detection of toxigenic-pathogenic in Vibrio cholerae.




Toxin, toxin-coregulated pili, and the toxR regulon are essential for Vibrio cholerae pathogenesis in humans

Results show for the first time the role of a specific pilus structure in colonization of the human intestine by V. cholerae O1 and exemplify the significance of a genetic regulon in pathogenesis.

Polymerase chain reaction for detection of the cholera enterotoxin operon of Vibrio cholerae

A set of oligonucleotide primers and amplification conditions for the polymerase chain reaction to detect the ctx operon of V. cholerae and related organisms were identical with those obtained by the DNA colony hybridization test with the polynucleotide probe.

Detection of Genes Encoding Cholera Toxin (CT), Zonula Occludens Toxin (ZOT), Accessory Cholera Enterotoxin (ACE) and Heat‐Stable Enterotoxin (ST) in Vibrio mimicus Clinical Strains

A significant association between O antigens and enterotoxic activities in V. mimicus clinical strains is suggested, and multifactorial virulence potentials of this human pathogen are clearly demonstrated.

Identification of Vibrio parahaemolyticus Strains at the Species Level by PCR Targeted to the toxR Gene

A toxR-targeted PCR protocol for the specific detection of V. parahaemolyticus was established, suggesting that toxR sequence variation may reflect the phylogenetic relationships of Vibrio species.

Rapid Method for Species-Specific Identification ofVibrio cholerae Using Primers Targeted to the Gene of Outer Membrane Protein OmpW

A one-step multiplex PCR assay for the simultaneous amplification of ompW and ctxAgenes should be of considerable value in the screening of both toxigenic and nontoxigenic V. cholerae strains of clinical as well as environmental origin.

Molecular Analysis of Vibrio cholerae O1, O139, non-O1, and non-O139 Strains: Clonal Relationships between Clinical and Environmental Isolates

Studies of clonality performed by using enterobacterial repetitive intergenic consensus sequence PCR, Box element PCR, amplified fragment length polymorphism (AFLP), and pulsed-fleld gel electrophoresis (PFGE) collectively indicated that the V. cholerae O1 and O139 strains had a clonal origin, whereas the non-O1, non- o139 strains belonged to the same group.

Genotypes Associated with Virulence in Environmental Isolates of Vibrio cholerae

ABSTRACT Vibrio cholerae is an autochthonous inhabitant of riverine and estuarine environments and also is a facultative pathogen for humans. Genotyping can be useful in assessing the risk of

The light organ symbiont Vibrio fischeri possesses a homolog of the Vibrio cholerae transmembrane transcriptional activator ToxR

A cross-hybridizing DNA fragment to Vibrio cholerae toxR was cloned from the nonpathogenic light organ symbiont Vibrio fischeri, and three proteins homologous to V. cholerae ToxR, ToxS, and HtpG were

Vibrio parahaemolyticus has a homolog of the Vibrio cholerae toxRS operon that mediates environmentally induced regulation of the thermostable direct hemolysin gene

Vp-ToxR and Vc-toxR share a strikingly similar function, i.e., direct stimulation at the transcriptional level of the gene encoding a major virulence determinant (enterotoxin) of a Vibrio species.