Open AccessLoop‐mediated isothermal amplification method for rapid detection of the toxic dinoflagellate Alexandrium , which causes algal blooms and poisoning of shellfish
Author(s) -
Wang Li,
Li Lin,
Alam M. J.,
Geng Yuhuan,
Li Zhiyong,
Yamasaki Shinji,
Shi Lei
Publication year - 2008
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2008.01074.x
Subject(s) - loop mediated isothermal amplification , paralytic shellfish poisoning , dinoflagellate , biology , algal bloom , amplicon , shellfish poisoning , saxitoxin , shellfish , sybr green i , alexandrium tamarense , marine toxin , microbiology and biotechnology , toxin , polymerase chain reaction , zoology , botany , gene , dna , ecology , genetics , fishery , aquatic animal , phytoplankton , nutrient , fish <actinopterygii>
The marine dinoflagellate genus Alexandrium includes a number of species that produce potent neurotoxins responsible for paralytic shellfish poisoning, which in humans may cause muscular paralysis, neurological symptoms and, in extreme cases, death. Because of the genetic diversity of different genera and species, molecular tools may help to detect the presence of target microorganisms in marine field samples. Here we employed a loop‐mediated isothermal amplification (LAMP) method for the rapid and simple detection of toxic Alexandrium species. A set of four primers were designed based upon the conserved region of the 5.8S rRNA gene of members of the genus Alexandrium . Using this detection system, toxic Alexandrium genes were amplified and visualized as a ladder‐like pattern of bands on agarose gels under isothermal condition within 60 min. The LAMP amplicons were also directly visualized by eye in the reaction tube by the addition of SYBR Green I. This LAMP assay was 10‐fold more sensitive than a conventional PCR method with a detection limit of 5 cells per tube when targeting DNA from Alexandrium minutum . The LAMP assay reported here indicates the potential usefulness of the technique as a valuable simple, rapid alternative procedure for the detection of target toxic Alexandrium species during coastal water monitoring.