African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12401

Full Length Research Paper

A rapid detection method on-site for Zygosaccharomyces based on loop-mediated isothermal amplification (LAMP) combined with a lateral flow dipstick (LFD)

Yang Liu
  • Yang Liu
  • Key laboratory of Microbiol technology and Bioinformatics of Zhejiang Province, Zhejiang Institute of Microbiology, Hangzhou 310012, China.
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Jun Wu
  • Jun Wu
  • Lin'an Center for Disease Control and Prevention, Lin'an, 311300, China.
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Li-Qing Chen
  • Li-Qing Chen
  • Hangzhou Huqingyu tang Traditional Chinese Medicine Mordernize Institute, Hangzhou 311100, China.
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Ya Shi
  • Ya Shi
  • Key laboratory of Microbiol technology and Bioinformatics of Zhejiang Province, Zhejiang Institute of Microbiology, Hangzhou 310012, China.
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Hong-Xia Zhao
  • Hong-Xia Zhao
  • Key laboratory of Microbiol technology and Bioinformatics of Zhejiang Province, Zhejiang Institute of Microbiology, Hangzhou 310012, China.
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Wei Chen
  • Wei Chen
  • Hangzhou Huqingyu tang Traditional Chinese Medicine Mordernize Institute, Hangzhou 311100, China.
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Hong Li
  • Hong Li
  • China National Accreditation Institute for Conformity Assessment, Beijing 100062, China.
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Lu-Ping He
  • Lu-Ping He
  • Key laboratory of Microbiol technology and Bioinformatics of Zhejiang Province, Zhejiang Institute of Microbiology, Hangzhou 310012, China.
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  •  Received: 23 February 2021
  •  Accepted: 22 April 2021
  •  Published: 31 May 2021

Abstract

The Zygosaccharomyces is notorious for its remarkable spoilage characteristics. In the present study, the visual and rapid identification of the genus Zygosaccharomyces was performed by a loop-mediated isothermal amplification (LAMP) assay using specific primers in Mini Dry Bath within 30 min at 65°C followed by a lateral flow dipstick (LFD) detectionThe sensitivity evaluation revealed LAMP-LFD assay with 1.0×101 copies/μL of Zygosaccharomyces DNA as its detection limit, which was the same as the methods of real-time quantitative PCR (qPCR) and conventional polymerase chain reaction (PCR). However, qPCR or PCR methods not only need to be performed in a specialist analytical laboratory with expensive equipment but also with risk of aerosol pollution. The LAMP-LFD assay had no cross-reactivity against 10 other yeast species and its specificity was 100%. A total of 25 Qiangli loquat Dew samples (17 bottles bulged and eight normal-appearing) were detected within 40 min with 100 and 92.0% accurately and specifically, when compared with the qPCR assay and the microbiology culture method, respectively. Therefore, the simple, fast, sensitive and low-cost LAMP-LFD assay is an effective and useful tool for the on-site identification of the genus Zygosaccharomyces.
 
Key words: Zygosaccharomyces, on-site detection, loop-mediated isothermal amplification (LAMP), lateral flow dipstick (LFD).