Detection of Fungi from an Indoor Environment using Loop-mediated Isothermal Amplification (LAMP) Method

TAKAKO NAKAYAMA; TAKASHI YAMAZAKI; AYAKA YO; KAZUYA TONE; MOHAMED MAHDI ALSHAHNI; RYUICHI FUJISAKI; KOICHI MAKIMURA

doi:10.4265/bio.22.97

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Detection of Fungi from an Indoor Environment using Loop-mediated Isothermal Amplification (LAMP) Method

TAKAKO NAKAYAMA, TAKASHI YAMAZAKI, AYAKA YO, KAZUYA TONE, MOHAMED MAHDI ALSHAHNI, RYUICHI FUJISAKI, KOICHI MAKIMURA

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Keywords: Environmental fungi, Detection of fungi, Loop-mediated isothermal amplification (LAMP), Procedure for Ultra Rapid Extraction (PURE)

JOURNAL FREE ACCESS

2017 Volume 22 Issue 2 Pages 97-104

DOI https://doi.org/10.4265/bio.22.97

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Abstract

　Loop-mediated isothermal amplification (LAMP) is a useful DNA detection method with high specificity and sensitivity. The LAMP reaction is carried out within a short time at a constant temperature without the need for thermal cycling. We developed a LAMP primer set for detecting a wide range of fungi by aligning the sequences of the large subunit ribosomal RNA gene of Candida albicans (Ascomycota), Cryptococcus neoformans (Basidiomycota), and Mucor racemosus (Mucorales). The threshold of C. albicans rDNA as template with our LAMP primer set was in the range of 10-100 copies per a reaction. In this study, we evaluated the correlation between colony forming units (CFU) and LAMP detection rate using the LAMP method for environmental fungi. The LAMP method should be a useful means of detecting fungi in indoor environments, disaster areas, or even in confined manned spacecraft to prevent allergies or infections caused by fungi.

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