MOLECULAR IDENTIFICATION AND CLONAL RELATION OF ATYPICAL ISOLATE Aeromonas salmonicida USING RESTRICTION FRAGMENT LENGTH POLYMORPHISM (RFLP)
Abstract: Aeromonas
salmonicida is responsible in many cases of furunculosis outbreaks resulting in
economic loss of freshwater aquaculture. Four isolates of A. salmonicida have
been isolated from fish collected in four different regions in Indonesia and
its clonal relation has yet to be determined. In the research, A. salmonicida
isolates and ATCC atypical isolate as the control had been checked for their
clonal relation using Restriction Fragment Length Polymorphism (RFLP) method in
which restriction enzyme of AluI, HaeIII, MboI, and EheI were used. PCR test
results using the primers 16S rDNA amplicon gave a positive response to the
1300 bp band. The result of RFLP analysis showed that A. salmonicida atypical
isolates from Indonesia represent subspecies smithia except isolates from C.
macropomum in Yogyakarta in MS and 16S rDNA regions. Isolates from Jambi,
Pontianak and Semarang showed a slight variation on enzyme restriction sites.
Isolates number 2, 3, 4 and 5 had the same restriction sites using AluI enzyme
with MS primer. The restriction enzymes that could give the best result for
RFLP method of A. salmonicida were HaeIII, MboI and EheI.
Keywords: A. salmonicida;
furunculosis; PCR; RFLP; restriction enzyme
Author: Riza Priyatna, Indaryulianto,
Kurniasih
Journal Code: pperikanangg110022
