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Development of Rapid and Specific Detection for the Human Aichivirus A Using the Loop-Mediated Isothermal Amplification from Water Samples

  • J. Y. Lee
  • J. H. Kim
  • J. Y. RhoEmail author
Short communications
  • 51 Downloads

Abstract

Human Aichivirus A (AiV-A) is classified as a Kobuvirus, group IV positive sense single strand RNA viruses. The first outbreak of AiV-A was reported from Aichi Prefecture, Japan in 1989. AiV-A exists not only among clinical patients, such as diarrhea, but also in a variety of water environments, as its occurrence is reported across a wide geographical range, from developing to advanced countries. For diagnose of AiV-A from water samples, mostly polymerase chain reaction (PCR) system have been developed. However, loop-mediated isothermal amplification (LAMP) assay has not been applied. In this study, developed a LAMP method to achieve a rapid, specific and highly sensitive detection of AiV-A. The method developed in this study is aimed specifically at AiV-A. Through a specific and non-specific selection and sensitivity test process for the five prepared LAMP primer sets, one primer set and optimum reaction temperature were selected. A newly developed method was more rapid (approximately 2–8 h), specific and equivalent detection of AiV-A than with the conventional PCRs. In addition, confirm system of positive LAMP reaction was developed by using the restriction enzyme Aci I and Hae III. For evaluation and verification of developing LAMP assay, a was applied to twenty cDNA from groundwater samples. This study proved rapid and specific diagnosis of AiV-A from water samples, and it is also demanded to be applicable to other environmental, clinical and food samples.

Keywords

AiV-A Human AichivirusLAMP Loop-mediated isothermal amplification Water sample 

Notes

Acknowledgements

This work was supported by the National Research Foundation of Korea (NRF) Grant funded by the Korea Government (MSIT) (No. 2017R1A2B1009303).

Supplementary material

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Supplementary material 1 (DOCX 14 kb)
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Supplementary material 3 (DOCX 19 kb)

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Copyright information

© Association of Microbiologists of India 2019

Authors and Affiliations

  1. 1.Department of Microbiology, College of Natural SciencesDankook UniversityCheonanKorea
  2. 2.Department of Chemistry, College of Natural SciencesDankook UniversityCheonanKorea
  3. 3.Institute of Tissue Regeneration Engineering (ITREN)Dankook UniversityCheonanKorea

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