Open AccessApplication of loop mediated isothermal amplification (LAMP) method for detection of Edwarsiella ictaluri on patin (Pangasius sp.) fish
Author(s) -
Hessy Novita,
Uni Purwaningsih,
Angela Mariana Lusiastuti
Publication year - 2021
Publication title -
iop conference series. earth and environmental science
Language(s) - English
Resource type - Journals
eISSN - 1755-1307
pISSN - 1755-1315
DOI - 10.1088/1755-1315/744/1/012050
Subject(s) - loop mediated isothermal amplification , pangasius , sybr green i , edwardsiella ictaluri , catfish , biology , orange (colour) , primer (cosmetics) , pathogen , fish <actinopterygii> , microbiology and biotechnology , veterinary medicine , polymerase chain reaction , dna , fishery , food science , gene , chemistry , genetics , medicine , organic chemistry , ictalurus
Edwarsiela ictaluri is a potential bacteria as a pathogen causing Enteric Septicemia of Catfish (ESC) and also still a big problem in catfish because it can cause death to 100%. This study aims to develop diagnosis DNA based on molecular using Loop Mediated Isothermal Amplification. Method (LAMP). Detection DNA of E. ictaluri bacteria from 9 isolates was extracted at Fish Health Laboratory, in Depok. used four pieces of specific primers from Eip 18 gene of E. ictaluri . DNA amplification, for LAMP reactions without a thermocycler machine, using only an incubator/water bath/oven, at 65°C for 1 hour. The result of detection with LAMP, showed that 8 isolates were positive of E. ictaluri marked by SYBR green I color change from orange to green. While 1 isolate was negatively marked with SYBR Green remain orange. The LAMP results are also verified with PCR using outer primer F3 and B3 which showed positive E. ictaluri . LAMP can detect ESC disease simple, quickly, easily, economically in 1 hour compared by PCR and LAMP method can be used as an alternative diagnostic test for fish diseases, especially in areas with limited laboratory infrastructure.