Detection of representative enteropathogenic bacteria, Vibrio spp., pathogenic Escherichia coli, Salmonella spp., Shigella spp., and Yersinia enterocolitica, using a virulence factor gene-based oligonucleotide microarray

Kim, Dong-Hun; Lee, Bok-Kwon; Kim, Yong‐Dae; Rhee, Sung‐Keun; Kim, Young-Chang

doi:10.1007/s12275-010-0119-5

Cited by 17 publications

(15 citation statements)

References 35 publications

(66 reference statements)

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“…Oligonucleotide‐based microarrays using molecular PCR methods have been developed that rapidly detect rRNA and virulence‐factor genes in food, soil and water. These techniques are also sensitive enough to detect the major gastrointestinal bacteria in human stool samples . The major aetiological agents of parasitic diarrhoea are Cryptosporidium spp., Entamoeba histolytica and Giardia liamblia .…”

Section: Introductionmentioning

confidence: 99%

Simultaneous detection of gastrointestinal pathogens with a multiplex Luminex-based molecular assay in stool samples from diarrhoeic patients

Mengelle¹,

Mansuy²,

Prère

et al. 2013

Clinical Microbiology and Infection

117

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We have evaluated the multiplex molecular method xTAG(®) Gastrointestinal Panel (GPP) for detecting pathogens in stool samples of diarrhoeic patients. We collected 440 samples from 329 patients (male:female ratio of 1.2:1), including 102 immunosuppressed adults, 50 immunosuppressed children, 56 children attending the neonatal unit and 121 children attending the emergency unit. Of these, 176 samples from 162 patients were xTAG(®) GPP positive (102 viruses, 61 bacteria and 13 parasites) and the assay was more sensitive than the conventional test for detecting rotavirus (p <0.01), noroviruses (p <0.0001), Salmonella spp. (p <0.001), Campylobacter spp. (p <0.001) and toxigenic Clostridium difficile (p 0.005). The predominant pathogens were viruses (23.2%), with rotavirus (15.9%) being the most common. Bacterial agents were detected in 13.9%; the most common was Salmonella spp. (4.8%). Parasites were detected in 2.9%; Cryptosporidium spp. (2%) was the most common. There were 31 co-infections (7% of samples), involving two pathogens in 23 (5.2%) and three pathogens in eight (1.8%) samples. There were 113 (92.6%) positive samples from the children attending the emergency unit, 25 (17%) positive samples from immunosuppressed adults, 22 (25.3%) positive samples from immunosuppressed children and 16 (19%) positive samples from children attending the neonatal unit. The low turnaround time and technical hands-on time make this multiplex technique convenient for routine use. Nevertheless, conventional bacterial culture and parasitological stool examination are still required to detect other pathogens in specific cases and to determine susceptibility to antibiotics.

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Section: Introductionmentioning

confidence: 99%

Simultaneous detection of gastrointestinal pathogens with a multiplex Luminex-based molecular assay in stool samples from diarrhoeic patients

Mengelle¹,

Mansuy²,

Prère

et al. 2013

Clinical Microbiology and Infection

117

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“…It is thus imperative to include sufficient numbers of negative and internal positive controls [21]. Microarray is another method, which is quite sensitive and effective for identifying the target microbe [9,[22][23][24]. A more recent DNA based diagnostic tool for identifying food borne pathogens method is the Loop-mediated isothermal amplification (LAMP).…”

Section: Molecular Methodsmentioning

confidence: 99%

Genome Wide Search for Biomarkers to Diagnose Yersinia Infections

Kalia

Kumar

2015

Indian J Microbiol

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Bacterial identification on the basis of the highly conserved 16S rRNA (rrs) gene is limited by its presence in multiple copies and a very high level of similarity among them. The need is to look for other genes with unique characteristics to be used as biomarkers. Fifty-one sequenced genomes belonging to 10 different Yersinia species were used for searching genes common to all the genomes. Out of 304 common genes, 34 genes of sizes varying from 0.11 to 4.42 kb, were selected and subjected to in silico digestion with 10 different Restriction endonucleases (RE) (4-6 base cutters). Yersinia species have 6-7 copies of rrs per genome, which are difficult to distinguish by multiple sequence alignments or their RE digestion patterns. However, certain unique combinations of other common gene sequences-carB, fadJ, gluM, gltX, ileS, malE, nusA, ribD, and rlmL and their RE digestion patterns can be used as markers for identifying 21 strains belonging to 10 Yersinia species: Y. aldovae, Y. enterocolitica, Y. frederiksenii, Y. intermedia, Y. kristensenii, Y. pestis, Y. pseudotuberculosis, Y. rohdei, Y. ruckeri, and Y. similis. This approach can be applied for rapid diagnostic applications.

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“…You et al combined a multiplex PCR of virulence gene targets of eight enteropathogenic bacteria with DNA microarray detection with a comparable LOD of 58 cfu/ml [36]. A DNA microarray for the detection of 10 bacterial pathogens was developed by Kim et al [28]. Here, the microarray detection was combined with multiplex PCR of bacterial specific virulence factors.…”

Section: Infectious Diarrheamentioning

confidence: 99%