scholarly journals Short Communication: Detection of Salmonella typhimurium ATCC 14028 and Listeria monocytogenes ATCC 7644 in processed meat products using Real-Time PCR Multiplex Method

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Alfi Sophian ◽  
RATNA PURWANINGSIH ◽  
EKA PUTRI JUNIARTI IGIRISA ◽  
MUHAMMAD LUTHFI AMIRULLAH ◽  
BERTHA LOLO LUKITA ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Salmonella Typhimurium ◽  
Real Time ◽  
Melting Temperature ◽  
Real Time Pcr ◽  
Meat Products ◽  
Processed Meat ◽  
Ct Value ◽  
Pcr Multiplex ◽  
Pcr Method

Abstract. Sophian A, Purwaningsih R, Igirisa RPJ, Amirullah ML, Lukita BL, Fitri RA. 2020. Short Communication: Detection of Salmonella typhimurium ATCC 14028 and Listeria monocytogenes ATCC 7644 in processed meat products using Real-Time PCR Multiplex Method. Asian J Nad Prod Biochem 21: 17-20. The detection of Salmonella typhimurium ATCC 14028 and Listeria monocytogenes ATCC 7644 in processed meat products was carried out using Multiplex Real-Time PCR (qPCR) in the Microbiology and Molecular Biology Laboratory at the Indonesian Food and Drug Authority in Gorontalo. The purpose of this study was to provide alternative testing methods for food products circulating in the market. The sample consisted of 25 samples of processed meat products spike with Salmonella typhimurium ATCC 14028 phase 2 and Listeria monocytogenes ATCC 7644 phase 2. The method used in the study was qPCR analysis using the SYBR Green method, while DNA isolation used the direct PCR method. Data analysis was carried out based on Cycle threshold and Melting temperature based on two main criteria. Cycle threshold (Ct) analysis determines the Ct value of the sample and comparing it with the control. Melting temperature (Tm) analysis determines the temperature at which 50% of double-stranded DNA changed to a single standard and comparing it with the melting temperature of positive control. The results showed Salmonella typhimurium ATCC 14028 in the processed meat was detected at an average Ct value of 10.34, and a Tm value of 85.70. The presence of Listeria monocytogenes ATCC 7644 in the samples was recognized at an average Ct value of 14.04, and an average Tm value of 80.07. It can be concluded that the real-time multiplex PCR method can be used to detect Salmonella typhimurium ATCC 14028 and Listeria monocytogenes ATCC 7644 by using the melting curve (Tm) analysis.

scholarly journals Methods of Detecting Meat Species in Food of Animal Origin

2018 ◽  
Vol 75 (2) ◽  
pp. 125
Author(s):  
Lavinia-Maria CHIŞ ◽  
Dan-Cristian VODNAR
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Meat Product ◽  
Meat Products ◽  
Ease Of Use ◽  
Animal Origin ◽  
Processed Meat ◽  
Elisa Method ◽  
Food Of Animal Origin ◽  
Pcr Method

An important factor in the detection of falsification is the control of the composition of the meat at each stage of manufacturing the product. The PCR method is based on the study of proteins and meat nucleic acids used in food for the detection of animal species. Another technique is the Elisa method that works on the principle of identification and measurement of the quantity of molecules in a sample. There are several types of Elisa to increase specificity due to differences in structure and sample characteristics. By comparing the two methods used to identify the processed meat product species, Real Time PCR had the highest prediction as results. However, the Elisa method is more time efficient and easier to use. Real Time PCR is effective in identifying processed meat products that require low detection. The Elisa Kit is useful because of the ease of use.

scholarly journals Real-Time PCR Method Combined with a Matrix Lysis Procedure for the Quantification of Listeria monocytogenes in Meat Products

Foods ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 735
Author(s):  
Mirian Labrador ◽  
Carlota Giménez-Rota ◽  
Carmen Rota
Keyword(s):  
Listeria Monocytogenes ◽  
Real Time ◽  
Real Time Pcr ◽  
Foodborne Pathogen ◽  
Meat Products ◽  
Reference Method ◽  
Meat Industry ◽  
Gene Copies ◽  
Pcr Method ◽  
Meat Samples

In this study a real-time PCR method has been developed for the specific quantification of the foodborne pathogen Listeria monocytogenes on meat products through the gene hlyA. The PCR was combined with a matrix lysis that allowed the obtaining of the microorganisms without sample dilution and the elimination the PCR inhibitors from dry-cured ham. The qPCR method calibration curve had an efficiency of 100.4%, limits of detection and quantification were 30.1 ± 6.2 CFU/g which is under the legal limit of L. monocytogenes in ready-to-eat products, and an analytical variability <0.25 log hlyA gene copies/reaction. The analysis was performed simultaneously with the reference method ISO 11290-2. The comparison of the qPCR-matrix lysis results with the reference method showed an excellent correspondence, with a relative accuracy between 95.83–105.20%. Finally, the method was applied to commercial derived meat samples and the pathogen was quantified in one of the commercial samples assayed in 69.1 ± 13.9 CFU/g while the reference method did not quantify it. The optimized qPCR showed higher precision and sensitivity than the reference method at low concentrations of the microorganism in a shorter time. Therefore, qPCR-matrix lysis shows a potential application in the meat industry for L. monocytogenes routine control.

scholarly journals Epidemiological survey on the prevalence of Salmonella spp. in the Sardinian pig production chain, using real-time PCR screening method

2019 ◽  
Vol 8 (2) ◽  
Author(s):  
Carlo Pala ◽  
Tiziana Tedde ◽  
Sara Salza ◽  
Maria Teresa Uda ◽  
Stefano Lollai ◽  
...  
Keyword(s):  
Salmonella Typhimurium ◽  
Multiplex Pcr ◽  
Real Time ◽  
Real Time Pcr ◽  
Meat Products ◽  
Processed Meat ◽  
Finishing Pigs ◽  
Salmonella Spp ◽  
Production Chain ◽  
Pig Production

The aim of this study was to evaluate the prevalence of Salmonella spp. in the Sardinian pig production chain in order to establish the incidence of monophasic serovariant of Salmonella Typhimurium on isolates with molecular methods (real-time PCR and multiplex PCR). Samples were collected in three EC slaughterhouses, four small slaughterhouses annexed to farmhouses, one meat distribution center, four meat cutting laboratories and four sausage processing plants. A total of 166 samples were collected and analyzed: 46 environmental samples, 48 finishing pigs, 16 piglets, 24 samples of non-processed meat, 28 meat preparations and 4 meat products. All samples were processed with an initial screening using the real-time PCR MicroSEQ® Salmonella spp detection Kit (Applied biosystems, life technologies) and with the TaqMan® Real-time PCR to confirm the kit results. Samples that tested positive for Salmonella spp were confirmed with cultural method using the standard ISO 6579. Positive samples were submitted to phenotypic identification. One colony from each positive sample was serotyped with multiplex PCR method. Salmonella spp was isolated in 7 on 166 samples (4.22 %). Among the positive samples, two came from finishing pigs, two belonged to the category meat preparations, two to meat products, one was an environmental sample. Multiplex PCR confirmed that the collected strains belonged to the species Salmonella Typhimurium (1), Salmonella derby (3) and monophasic serovariant of Salmonella Typhimurium (3).

A Fast and Reliable Real-Time PCR Method for Detection of Ten Animal Species in Meat Products

2018 ◽  
Vol 83 (2) ◽  
pp. 258-265 ◽  
Author(s):  
Lissandra Sousa Dalsecco ◽  
Rafael Melo Palhares ◽  
Pollyana Carvalho Oliveira ◽  
Lilian Viana Teixeira ◽  
Marcela Gonçalves Drummond ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Animal Species ◽  
Meat Products ◽  
Pcr Method

scholarly journals Detection of meat from horse, donkey and their hybrids (mule/hinny) by duplex real-time fluorescent PCR

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0237077
Author(s):  
Dan Wang ◽  
Liping Wang ◽  
Chenyu Xue ◽  
Yuebei Han ◽  
Hejing Li ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Limit Of Detection ◽  
Meat Products ◽  
Processed Meat ◽  
Similar Species ◽  
Fluorescent Pcr ◽  
Species Specific ◽  
Muscle Gene ◽  
Fluorescence Amplification

Meat adulteration is currently a common practice worldwide. In China, adulteration of donkey meat products with the similar species (horse and mule/hinny) meat and mislabeling are becoming widespread concerns. In this study, a sensitive and species-specific duplex real-time PCR assay based on the simultaneous amplification of fragments of the creatine kinase muscle gene family, was developed and optimized for the identification of horse, donkey and mule /hinny species in raw and heat-processed meat products. Duplex real-time PCR results showed different fluorescence amplification curves for horse and donkey. Both kinds of fluorescence amplification curves appeared simultaneously for mule/hinny. The limit of detection (LOD) of the method was up to 0.01 ng /μL. The method and strategy developed in this study could be applied to detect the presence of adulterants from horse and mule /hinny meat in raw donkey meat and meat products.

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