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Endonuclease Restriction-Mediated Real-Time PCR for Simultaneous Detection of Listeria monocytogenes and Listeria ivanovii

Abstract

Listeria monocytogenes and Listeria ivanovii have been identified as the only two pathogenic species in Listeria genus. Food contamination with L. monocytogenes was a severe public health issues accompanied with great economic loss, while the contamination of L. ivanovii are uncommon but it represents a potential risk of infection in immunocompromised people. The sensitive and rapid method for detection of L. monocytogenes and L. ivanovii is beneficial to medical diagnosis and food industry. Here we developed an endonuclease restriction-mediated real-time PCR (ET-PCR) assay allowing for concomitant detection of L. monocytogenes and L. ivanovii. What make this novel PCR assay convenient in application were the simplicity of primer design and allowing for multi-targets detection. The special forward or reverse primer consisted of reporter dye, oligonucleotides containing the recognition site of restriction enzyme BstUI, oligonucleotides for PCR amplification and quenching dye. It will be achieved multi-targets analysis by restriction enzyme digestion and different reporter dyes release. As few as 80 femtogram target DNA per reaction can be detected by both single and duplex ET-PCR assay, which was 100-fold more sensitive than normal PCR assay. This duplex ET-PCR assay was able to discriminate L. monocytogenes and L. ivanovii from other common foodborne pathogens. Combined with enrichment, this novel assay offers a sensitive and feasible method for detection of L. monocytogenes and L. ivanovii from different matrices. It may be an efficient tool for epidemiologist and clinician in their effort to prevent and control the infections caused by L. monocytogenes or L. ivanovii.

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Publication details

The article was received on 17 Nov 2017, accepted on 06 Feb 2018 and first published on 06 Feb 2018


Article type: Paper
DOI: 10.1039/C7AY02667F
Citation: Anal. Methods, 2018, Accepted Manuscript
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    Endonuclease Restriction-Mediated Real-Time PCR for Simultaneous Detection of Listeria monocytogenes and Listeria ivanovii

    Y. Wang, Y. Wang and C. Ye, Anal. Methods, 2018, Accepted Manuscript , DOI: 10.1039/C7AY02667F

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