Taqman RT-PCR Analysis for the Identification of the Osmotolerant Yeast Zygosaccharomyces rouxii

Syromyatnikov, Mikhail; Savinkova, O. V.; Kiryanova, S. V.; Frolov, Evgenii N.; Popov, Vasily N.

doi:10.1134/s0003683818060133

Cited by 1 publication

(1 citation statement)

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“…Some studies have suggested that species or genus specific PCR primers can be used to identify the genus Zygosaccharomyces (Harrison et al, 2011;Hulin and Wheals, 2014), but missing LOD data. Moreover, there was very few paper detected Z. rouxii using the qPCR assay with LOD approximately 20 times more sensitive than the LAMP assay in the present study (Syromyatnikov et al, 2018). This may be attributed to the set of primers only applied to the specific Zygosaccharomyces spp.…”

Section: Liu Et Al 205contrasting

confidence: 51%

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

Liu¹,

Wu²,

Chen³

et al. 2021

Afr. J. Biotechnol.

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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) detection. The 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 crossreactivity 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.

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Section: Liu Et Al 205contrasting

confidence: 51%