Open AccessDetection of Salmonella typhimurium and Escherichia coli in artificially inoculated Milk sample using Real Time PCR method
Author(s) -
Muktiningsih Nurjayadi,
Ulfi Rahma Efrianti,
N. Azizah,
Fera Kurniadewi,
V. Saamia,
I. M. Wiranatha,
L. Nastassya
Publication year - 2019
Publication title -
journal of physics. conference series
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.21
H-Index - 85
eISSN - 1742-6596
pISSN - 1742-6588
DOI - 10.1088/1742-6596/1402/5/055084
Subject(s) - primer (cosmetics) , escherichia coli , melting curve analysis , salmonella , bacteria , detection limit , microbiology and biotechnology , biology , enterobacteriaceae , real time polymerase chain reaction , polymerase chain reaction , chemistry , food science , chromatography , gene , genetics , organic chemistry
Salmonella typhimurium and Escherichia coli are the bacteria the most common cause of food-borne illness. The aim of the study is to detection S. typhimurium and E. coli by Real Time PCR from milk samples. Gene targeted of S. typhimurium is pef and the E. coli is fim C. Confirmation test of all primers in contaminated milk with S. typhimurium and E. coli gave different cycle threshold (Ct) value in range 12-13. The detection limit for S. typhimurium with primer pef was in 7, 92 pg/μL and detection limit for E. coli with primer fim C in 7, 12 pg/μL. Each primer produces distinctive melting peak. The melting peak curve S. typhimurium with pef primer is 84,17°C and the melting peak curve E. coli with fim C primer is 79,97°C Based on the result of this study; all the primers were successfully applied in detecting S. typhimurium and E. coli bacteria in the milk sample quickly, sensitive and specific as well as can determine the number of bacteria accurately.