Establishment and application of a multiple cross displacement amplification combined with nanoparticles-based biosensor
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RESEARCH ARTICLE
Open Access
Establishment and application of a multiple cross displacement amplification combined with nanoparticles-based biosensor method for the detection of Bordetella pertussis Shijun Li1†, Chunting Liu1†, Ying Liu1, Qing Ma1, Yue Wang1 and Yi Wang2,3*
Abstract Background: Bordetella pertussis is the causative agent of pertussis, a respiratory tract infectious disease. Efficient techniques for detection of B. pertussis isolates are important for clinical diagnosis. Multiple cross displacement amplification (MCDA), a novel isothermal amplification based molecular detection method, has been developed to overcome the technical drawback of the current methods in recent years. This aim of this study is to develop a MCDA with Nanoparticles-based Lateral Flow Biosensor (MCDA-LFB) for the detection of B. pertussis. Results: A set of 10 primers based on the pertussis toxin (PT) promoter region sequence of B. pertussis was designed. The B. pertussis-MCDA-LFB assay was successfully established and optimized at 64 °C for reaction of 40 min. The detection limit was determined as 10 fg/reaction of pure DNA, and no cross-reactions to non-B. pertussis strains were observed, based on the specificity validation. The whole operation, ranging from template preparation to result reporting, could be completed within 70 min without requirement of costly equipment. The B. pertussis-MCDA-LFB in clinic sample detection yielded identical positive rates with traditional culture and showed higher sensitivity than conventional PCR. The results of MCDA-LFB are easier to read due to the usage of LFB. Conclusions: The isothermal amplification based MCDA-LFB established in the present study is a specific, sensitive, rapid and economical technique for the detection of B. pertussis. Keywords: Bordetella pertussis, Multiple cross displacement amplification, Lateral flow biosensor, MCDA- LFB, Detection limit
* Correspondence: [email protected] † Shijun Li and Chunting Liu contributed equally to this work. 2 Department of Respiratory Disease, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health, Beijing 10045, PR China 3 Key Laboratory of Major Diseases in Children, Ministry of Education, Beijing Key Laboratory of Pediatric Respiratory Infection Disease, National Clinical Research Center for Respiratory Diseases, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health, Beijing 10045, PR China Full list of author information is available at the end of the article © The Author(s). 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in
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