Development and application of an oligonucleotide microarray for the detection of food-borne bacterial pathogens

Xinwei, Wang; Zhang, Liang; Jin, Lianqun; Jin, Min; Shen, Zhiqiang; An, Shuang; Chao, Fuhuan; Lǐ, Jùnwén

doi:10.1007/s00253-007-0993-x

Cited by 84 publications

(47 citation statements)

References 41 publications

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“…DNA arrays have two main uses in microbiology: Detection of known pathogens, and assessment of diversity 113) . In general, a PCR amplification step using specific primer sets is used to generated labeled DNA 9,11,27,58,79,95,96,100,105,136,156,173,182) . This requirement of predesigned oligomers for amplification and hybridization opens the possibility that many unknown organisms remain undetected (especially viruses which have no universal homologous gene).…”

Section: Introductionmentioning

confidence: 99%

Global Sequencing: A Review of Current Molecular Data and New Methods Available to Assess Microbial Diversity

Christen

2008

Microb. Environ.

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Roughly estimating the diversity of life has for a long time been seen as a challenge. Sequences obtained from the environment by direct amplification are now seen as the sole mean to provide information for at least 99% of the prokaryotes in natural communities. Recent massive parallel sequencing technologies now allow to sequence nearly every sequence resulting from such amplification. Are we ready to deal with such a deluge of data? Using three recent publications (16S rRNA genes amplification in soil and seawater followed by massive parallel sequencing) as case examples, this review analyses the state of the field.

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Section: Introductionmentioning

confidence: 99%

Global Sequencing: A Review of Current Molecular Data and New Methods Available to Assess Microbial Diversity

Christen

2008

Microb. Environ.

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“…Wang et al [62] presented an array-based assay for the identification of 23 foodborne pathogens. Their array consists of 20-30mers spotted on a glass slide, and is designed to hybridize to the 16S gene of target pathogens.…”

Section: Microarray-based Diagnosticsmentioning

confidence: 99%

Potentials and limitations of molecular diagnostic methods in food safety

Lauri

Mariani

2008

Genes Nutr

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Molecular methods allow the detection of pathogen nucleic acids (DNA and RNA) and, therefore, the detection of contamination in food is carried out with high selectivity and rapidity. In the last 2 decades molecular methods have accompanied traditional diagnostic methods in routine pathogen detection, and might replace them in the upcoming future. In this review the implementation in diagnostics of four of the most used molecular techniques (PCR, NASBA, microarray, LDR) are described and compared, highlighting advantages and limitations of each of them. Drawbacks of molecular methods with regard to traditional ones and the difficulties encountered in pathogen detection from food or clinical specimen are also discussed. Moreover, criteria for the choice of the target sequence for a secure detection and classification of pathogens and possible developments in molecular diagnostics are also proposed.

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“…The oligonucleotide probes were fixed on the membrane, and cross linked by UV crosslinker for 30 seconds to allow binding of probes onto the nylon membrane. After cross linking, any unbounded oligonucleotides were removed by two times washing in 0.5x SSC, 0.1% sodium dodecyl sulfate (SDS) for two minutes at 37˚C (5,11). The strips were dried and stored at room temperature.…”

Section: Preparation Of the Oligonucleotide Arraymentioning

confidence: 99%

“…Therefore, the identification of pathogens greatly depends on the help provided by clinical laboratories (3,4). There are several traditional microbiological methods to detect pathogenic bacteria, which rely on culture followed by standard biochemical and serological methods (5,6). These methods are very time consuming, onerous and not sensitive enough.…”

Section: Introductionmentioning

confidence: 99%

“…Miller et al (18) reported that microarray technology and its application to diagnose infectious diseases, is vastly grew from 2000 to 2008. Among the rDNA genes in bacterial genomes, the 23S rDNA genes are the most frequently used markers and are repeatedly applied to detect and identify intestinal pathogens by DNA microarray (5,7,12). Wilson et al (19) described the multi pathogen identification (MPID) microarray for high confidence identification of eighteen pathogens.…”

Section: Introductionmentioning

confidence: 99%

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Simultaneous Detection of Escherichia coli, Salmonella enterica, Listeria monocytogenes and Bacillus cereus by Oligonucleotide Microarray

Sarshar¹,

Shahrokhi²,

Ranjbar³

et al. 2015

Int J Enteric Pathog

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Background:Traditional laboratory methods to detect pathogenic bacteria are time consuming and laborious. Therefore, it is essential to use powerful and reliable molecular methods for quick and simultaneous detection of microbial pathogens. Objectives: The current study aimed to evaluate the capability and efficiency of 23S rDNA sequence for rapid and simultaneous detection of four important food-borne pathogens by an oligonucleotide microarray technique. Materials and Methods:The 23S rDNA sequences of Escherichia coli, Salmonella enterica, Listeria monocytogenes and Bacillus cereus were obtained from GenBank databases and used to design the oligonucleotide probes and primers by Vector NTI software. Oligonucleotide probes were placed on a nylon membrane and hybridization was performed between probes and 23S rDNA digoxigenin-labeled polymerase chain reaction (PCR) products. Hybridization signals were visualized by NBT/BCIP color development. Results: Positive hybridization color was produced for Escherichia coli, Salmonella enterica, Listeria monocytogenes and Bacillus cereus. The oligonucleotide microarray detected all bacterial strains in a single reaction in less than five hours. The sensitivity of the performed microarray assay was 10 3 cfu/mL for each species of pathogen. No cross reaction was found between the tested bacterial species. Conclusions: The obtained results indicated that amplification of 23S rDNA gene followed by oligonucleotide microarray hybridization is a rapid and reliable method to identify and discriminate foodborne pathogens tested under the study.

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Development and application of an oligonucleotide microarray for the detection of food-borne bacterial pathogens

Cited by 84 publications

References 41 publications

Global Sequencing: A Review of Current Molecular Data and New Methods Available to Assess Microbial Diversity

Global Sequencing: A Review of Current Molecular Data and New Methods Available to Assess Microbial Diversity

Potentials and limitations of molecular diagnostic methods in food safety

Simultaneous Detection of Escherichia coli, Salmonella enterica, Listeria monocytogenes and Bacillus cereus by Oligonucleotide Microarray

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