Sensitive and rapid detection of Prymnesium parvum (Haptophyceae) by loop-mediated isothermal amplification combined with a lateral flow dipstick
Aquaculture(2019)
摘要
The Prymnesium parvum is a common nearshore harmful alga that secretes hemolytic ichthyotoxin, which affects gills of shellfish and induce death, thereby resulting in severe economic losses of aquaculture. In this study, the loop-mediated isothermal amplification (LAMP) was employed in combination with the lateral flow dipstick (LFD) visual detection method to establish a rapid LAMP-LFD detection method for P. parvum. Approximately 50 min of LAMP amplification was necessary to generate the LFD results, which was about 2.5 h shorter than that of conventional PCR. Moreover, the LAMP-LFD method had excellent specificity, as indicated by the negative results for other common harmful algal species. It could detect genomic DNA at the lowest concentration of 3 × 10−2 ng μL−1. Therefore, the LAMP-LFD method could specifically and effectively detect target ITS sequence (including the 5.8S rDNA) of P. parvum, which is relatively simple and does not require specialized instruments. Consequently, it could be used as a rapid field detection method for the detection of low-density P. parvum as well as for the prevention of microalgal blooms.
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关键词
LAMP,LFD,ITS,PCR,P.P,AGE,FITC,FIP,BIP,POCT
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