Development of a loop-mediated isothermal amplification assay for the rapid detection of Russula subnigricans and Russula japonica
Russula subnigricans is the only deadly species in the genus Russula with a mortality rate of more than 50%, and Russula japonica is the most common poisonous species, making rapid species identification in mushroom poisoning incidents extremely important. The main objective of this study was to dev...
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Published in | Frontiers in microbiology Vol. 13; p. 918651 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
Frontiers Media S.A
23.08.2022
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Subjects | |
Online Access | Get full text |
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Summary: | Russula subnigricans
is the only deadly species in the genus
Russula
with a mortality rate of more than 50%, and
Russula japonica
is the most common poisonous species, making rapid species identification in mushroom poisoning incidents extremely important. The main objective of this study was to develop a rapid, specific, sensitive, and simple loop-mediated isothermal amplification (LAMP) assay for the detection of
R. subnigricans
and
R. japonica
. Two sets of species-specific LAMP primers targeting internal transcribed spacer (ITS) regions were designed to identify
R. subnigricans
and
R. japonica
. The results demonstrated that while LAMP could specifically detect
R. subnigricans
and
R. japonica
, the polymerase chain reaction (PCR) could not distinguish
R. subnigricans
from
Russula nigricans
. In addition, the results demonstrated that, compared to electrophoresis-LAMP and real-time quantitative LAMP (RT-qLAMP), the detection sensitivity of HNB-LAMP (a mixture of LAMP with hydroxy naphthol blue (HNB) dye) for
R. subnigricans
could reach 0.5 pg/μl and was 100-fold higher than that of PCR. The LAMP reaction could be completed in 45 min, which is much faster than the conventional PCR. In the future, LAMP can be used a quick, specific, and sensitive detection tool in various fields. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Reviewed by: Massimo Ferrara, Italian National Research Council, Italy; Biswajit Maiti, Nitte University, India This article was submitted to Food Microbiology, a section of the journal Frontiers in Microbiology Edited by: Giancarlo Perrone, Italian National Research Council, Italy |
ISSN: | 1664-302X 1664-302X |
DOI: | 10.3389/fmicb.2022.918651 |