Detection of eight foodborne bacterial pathogens by oligonucleotide array hybridization

Abstract

BackgroundSimultaneous and rapid detection of multiple foodborne bacterial pathogens is important for the prevention of foodborne illnesses.ObjectiveThe aim of this study was to evaluate the use of 16S rDNA and 23S rDNA sequences as targets for simultaneous detection of eight foodborne bacterial pathogens.MethodsNineteen bacterial oligonucleotide probes were synthesized and applied to nylon membranes. Digoxygenin labeled 16S rDNA and 23S rDNA from bacteria were amplified by PCR using universal primers, and the amplicons were hybridized to the membrane array. Hybridization signals were visualized by NBT/BCIP color development.ResultsThe eight intestinal bacterial pathogens including Salmonella enterica, Escherichia coli, Bacillus cereus, Vibrio cholerae, Shigella dysenteriae, Staphylococcus aureus, Listeria monocytogenes, and Enterococcus faecalis were appropriately detected in a panel of oligonucleotide array hybridization. The experimental results showed that the method could discriminate the bacterial pathogens successfully. The sensitivity of oligonucleotide array was 103 CFU/ml.ConclusionThis study showed that 16S rDNA and 23S rDNA genes had sufficient sequence diversity for species identification and were useful for monitoring the populations of foodborne pathogenic bacteria. Furthermore, results obtained in this study revealed that oligonucleotide array hybridization had a powerful capability to detect and identify the bacterial pathogens simultaneously.

Detection of 12 Common Food-Borne Bacterial Pathogens by TaqMan Real-Time PCR Using a Single Set of Reaction Conditions

“…In the work of Ishii et al microfluidic quantitative PCR (qPCR) technology was applied for the simultaneous detection and quantification of multiple food- and waterborne pathogens as L. monocytogenes, Salmonella Typhimurium, V. parahaemolyticus , and Clostridium perfringens (Ishii et al, 2013). Rapid and high-throughput identification of more food-borne bacterial pathogens by multiple analysis platforms have been developed and combination of several detection methods is also possible (Cremonesi et al, 2014; Salihah et al, 2016; Nasrabadi et al, 2017; Thomas et al, 2017; Ahn et al, 2018; Carloni et al, 2018; Zhang et al, 2018).…”

Simple Visualized Detection Method of Virulence-Associated Genes of Vibrio cholerae by Loop-Mediated Isothermal Amplification

“…To date, many methods have been developed for effective detection of O1/O139 V. cholerae contamination in food. Compared with the conventional culture-based microbiological detection assays, molecular biology-based methods are more rapid and sensitive, such as PCR (Kumar et al, 2010;Zago et al, 2017), real-time PCR (Garrido-Maestu et al, 2015;Casasola-Rodriguez et al, 2018), multiplex PCR (Bwire et al, 2018;Vu et al, 2018), oligonucleotide array hybridization (Nasrabadi et al, 2017), strand displacement amplification (SDA) (Phillips et al, 2018), rolling circle amplification (RCA) (Osterberg et al, 2014), cross-priming amplification (CPA) (Zhang et al, 2015), and nucleic acid sequence-based amplification (NASBA) (Fykse et al, 2012). Nevertheless, these methods require expensive equipments, which limit their wide application, particularly in on-site testing and large-scale survey.…”

Advances in the application of molecular diagnostic techniques for the detection of infectious disease pathogens (Review)

“…It is a high-throughput molecular diagnostic tool suitable for the detection of multiple infectious disease pathogens and drug resistance gene analysis ( 80 ). Nasrabadi et al ( 81 ) developed 16S and 23S rDNA-based probes able to simultaneously detect and identify eight food-borne bacterial pathogens. Ma et al ( 82 ) developed and evaluated a solid-phase chip for the simultaneous detection of 15 types of bacteria directly from respiratory tract specimens of patients with pneumonia, thus reducing detection time, facilitating the early administration of antimicrobial drugs and preventing bacterial resistance caused by empirical antibiotic treatment.…”