Detection of Salmonella typhimurium ATCC 14028 in supplement health product liquid preparation using Real-Time PCR (qPCR)

##plugins.themes.bootstrap3.article.main##

ALFI SOPHIAN
https://orcid.org/0000-0002-5206-2110
RATNA PURWANINGSIH
BERTHA LOLO LUKITA
ENI CAHYA NINGSIH

Abstract

Abstract. Sophian A, Purwaningsih R, Lukita BL, Ningsih EC. 2020. Detection of Salmonella typhimurium ATCC 14028 in supplement health product liquid preparation using Real-Time PCR (qPCR). Biofarmasi J Nat Prod Biochem 18: 65-69. Detection of Salmonella typhimurium ATCC 14028 using Real-Time PCR (qPCR) on health supplement products was carried out in the microbiology and molecular biology testing laboratory of the Food and Drug Supervisory Office in Gorontalo. The purpose of this study was to provide an alternative testing method reference in the testing of liquid supplement health supplement products in the market. The sample consisted of 35 samples of liquid supplement health supplements spike with positive control of Salmonella typhimurium ATCC 14028 phase 2. The method used in the study was qPCR analysis using the SYBR Green method, whereas DNA isolation using the direct PCR method. Data analysis was performed based on 2 main criteria: (i) Ct (Cycle threshold) analysis, which looks at the value of the sample Ct and compares it with controls, and (ii) analysis of melting temperature (Tm), which is the melting point at the temperature at which melting occurs and comparing the melting point to the positive control. The results showed that Salmonella typhimurium ATCC 14028 was detected in the sample at an average Ct value of 14.43 and an average Tm value of 86.05; for the specificity, LOD and positive control tests were all amplified. For negative controls, Ct and Tm values ??were not detected. Based on these data, it can be concluded that real-time PCR (qPCR) can be used to detect Salmonella typhimurium ATCC 14028 in liquid supplement health supplement products.

##plugins.themes.bootstrap3.article.details##

References
Piknova L, Kaclikova L, Pangallo D, Polek B, Kuchta T. 2005. Quantification of Salmonella by 5’-nuclease real-time polymerase chain reaction targeted to fimC gene. Curr. Microbiol. 50: 38-42.
Malorny B, Hoorfar J, Bunge C, Helmuth R. 2003. Multicenter Validation of the Analytical Accuracy of Salmonella PCR: towards an International Standard. Applied and Environmental Microbiology, 69: 290-296.
Catarame, T. M. G. 2006. Comparison of a real-time polymerase chain reaction assay with a culture method for the detection of Salmonella in retail meat samples. J. Food Saf. 26:1–15.
Wang X, Jothikumar M, Griffiths W. 2004. Enrichment and DNA extraction protocols for the simultaneous detection of Salmonella and Listeria monocytogenes in raw sausage meat with multiplex real-time PCR. J. Food Prot. 67:189–192.
Mercanoglu B, Griffiths MW. 2005. Combination of immunomagnetic separation with real-time PCR for rapid detection of Salmonella in milk, ground beef, and alfalfa sprouts. J. Food Prot. 68: 557–561.
Jothikumar N, Wang X, Griffiths M W. 2003. Real-time multiplex SYBR green I-based PCR assay for simultaneous detection of Salmonella serovars and Listeria monocytogenes. J. Food Prot. 66: 2141–2145.
Perelle S, F. Dilasser, Malorny B, Grout J, Hoorfar J, Fach P. 2004. Comparison of PCR-ELISA and LightCycler real-time PCRmassays for detecting Salmonella spp. in milk and meat samples. Mol. Cell. Probes 18:409–420.
Cheung, PY, Chan CW, Wong W, Cheung TL, Kam KM. 2004. Evaluation of two real-time polymerase chain reaction pathogendetection kits for Salmonella spp. in food. Lett. Appl. Microbiol. 39:509–515.
Ellingson JL, Anderson JL, Carlson SA, Sharma VK. 2004. Twelve-hour real-time PCR technique for the sensitive and specific detection of Salmonella in raw and ready-to-eat meat products. Mol. Cell. Probes 18:51–57.
Eyigor A, Carli KT. 2003. Rapid detection of Salmonella from poultry by real-time polymerase chain reaction with fluorescent hybridization probes. Avian Dis. 47:380–386.
Eyigor A, Carli KT, Unal CB. 2002. Implementation of real-time PCR to tetrathionate broth enrichment step of Salmonella detection in poultry. Lett. Appl. Microbiol. 34:37–41.
Oliveira ACS, Matheus C Rosa, Jéssica LB, Yasmine AM, Milena MAF, Gabrielle Virgínia FC, Andréia SS, Roberta SS, Josyane BS, Fábio FL, Talita BR, Carina M. 2018. Validating the Efficiency of a Simplex PCR and Quantitative SYBR Green qPCR for the Identification of Salmonella spp. DNA. J Food Microbiol Saf Hyg. 3:1.
BPOM. 2019. Regulation of the Food and Drug Supervisory Agency Number 16 of 2019. Concerning Supervision of Health Sumplemen. Republic of Indonesia Food and Drug Supervisory Agency. Jakarta. Indonesia. [Indonesian].
Jawetz E, Melnick JL, Adelberg EA. 2005, Medical Microbiology, translated by Mudihardi, E., Kuntaman, Wasito, EB, Mertaniasih, NM, Harsono, S., Alimsardjono, L., XXII Edition, 327-335, 362-363, Salemba Publishers Medika, Jakarta. [Indonesian].
Bohaychuck VM, Gensler GE, McFall ME, King RK, Renter DG. 2006. A Real-Time PCR Assay for the Detection of Salmonella in a Wide Variety of Food and Food-Animal Matrices. Journal of Food Protection, Vol. 70, No. 5, 2007, Pages 1080–1087.
Maddocks S, Olma T, dan Chen S. 2002. Comparison of CHROM agar Salmonella Medium and Xylose-Lysine-Desoxycholate and Salmonella-shigella agars for isolation of Salmonella Strains from stool samples. J. Clin. Microbiol. 40 (8): 2999-3003.
Nye KJ, Fallon D, Frodsham D, Geel B, Howel S, Messer S,Turner T, Warren RE. 2002. An evaluation of the performance of XLD, DCA, MLCB, and ABC agars as direct plating media for the isolation of Salmonella enterica from faeces. J Clin Pathol. 55: 286-288.
Baird R, Curtis G, dan Corry J. 1995. Culture Media for Food Microbiology. Volume 34. Amsterdam (NL). Elsevier Science B. V.
BAM. 2007. Bacteriological Analytical Manual. Chapter 5. Salmonella. December 2007 Content current as of June 18, 2009 .
Murray,P.R., Baron,E.J., Pfaller,M.A., Tenover,F.C. and Yolken,R.H. 1999. Manual of Clinical Microbiology, ASM Press, Washington D.C. USA.