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

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).

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

Asian Journal of Natural Product Biochemistry ◽

10.13057/biofar/f190103 ◽

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.

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Development and validation of two triplex real-time PCR systems for the simultaneous detection of six cereal species in processed meat products

Food Control ◽

10.1016/j.foodcont.2019.02.025 ◽

2019 ◽

Vol 101 ◽

pp. 180-188

Author(s):

K. Dolch ◽

M. Judas ◽

F. Schwägele ◽

D.A. Brüggemann

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Simultaneous Detection ◽

Meat Products ◽

Processed Meat ◽

Cereal Species ◽

Development And Validation

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

PLoS ONE ◽

10.1371/journal.pone.0237077 ◽

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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Development of a real-time PCR assay for the identification and quantification of bovine ingredient in processed meat products

Scientific Reports ◽

10.1038/s41598-020-59010-6 ◽

2020 ◽

Vol 10 (1) ◽

Cited By ~ 1

Author(s):

Xiaoyu Chen ◽

Lixia Lu ◽

Xiaohui Xiong ◽

Xiong Xiong ◽

Yuanjian Liu

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Meat Products ◽

Processed Meat ◽

Pcr Assay ◽

Identification And Quantification

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Methods of Detecting Meat Species in Food of Animal Origin

Bulletin of University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca Food Science and Technology ◽

10.15835/buasvmcn-fst:2018.0009 ◽

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.

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Detection and Quantification of Milk Ingredients as Hidden Allergens in Meat Products by a Novel Specific Real-Time PCR Method

Biomolecules ◽

10.3390/biom9120804 ◽

2019 ◽

Vol 9 (12) ◽

pp. 804 ◽

Cited By ~ 4

Author(s):

Caterina Villa ◽

Joana Costa ◽

Isabel Mafra

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Limit Of Detection ◽

Meat Products ◽

Milk Proteins ◽

12S Rrna ◽

Processed Meat ◽

Rrna Gene ◽

Wide Range ◽

Detection And Quantification

Milk ingredients are often included in a wide range of meat products, such as cooked hams and sausages, to improve technological characteristics. However, milk proteins are also important food allergens. The aim of this study was the development of a highly sensitive and specific real-time PCR system targeting the 12S rRNA gene of Bos domesticus for the detection and quantification of milk as an allergenic ingredient in processed meat products. The method was able to achieve an absolute limit of detection (LOD) of 6 fg of milk DNA. Using a normalized approach (∆Ct method) for the detection of milk protein concentrate (MPC), it was possible to obtain sensitivities down to 0.01% (w/w) of MPC in model hams (raw and cooked) and autoclaved sausages, and 0.005% in raw sausage mixtures. The developed systems generally presented acceptable PCR performance parameters, being successfully validated with blind samples, applied to commercial samples, and further compared with an immunochemical assay. Trace amounts of milk material were quantified in two out of 13 samples, but the results mostly infer the excessive practice of the precautionary labeling.

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Determination of the prevalence of Salmonella spp. and S. aureus in meat products by Real-Time PCR and testing their antibiotic susceptibility

Medycyna Weterynaryjna ◽

10.21521/mw.6533 ◽

2021 ◽

Vol 77 (06) ◽

pp. 6533-2021

Author(s):

REYHAN IRKIN ◽

BERKAY BOZKURT ◽

GULENDAM TUMEN

Keyword(s):

Public Health ◽

Staphylococcus Aureus ◽

Risk Factor ◽

Real Time ◽

Real Time Pcr ◽

Meat Products ◽

Penicillin G ◽

Diffusion Method ◽

Salmonella Spp ◽

And Staphylococcus Aureus

From a public health point of view meat products contain high pathogenic risk factors. This is because they can be contaminated with Salmonella spp. and Staphylococcus aureus microorganisms, and, more importantly, antibiotic resistance has been reported in these microorganisms at an increasing frequency. To examine the presence of Salmonella spp. and Staphylococcus aureus in samples of raw and semi-cooked (chicken doner, meat doner, chicken, beef, and lamb products) meat from markets of Izmir and Balikesir, Turkey were analysed. The presence of microorganisms in the samples was determined by Real-Time PCR method using Salmonella spp. and S. aureus specific primers. Following Real-Time PCR, microorganisms were isolated by selective culture methods and biochemical tests from the positive meat samples and tested for their antibiotic susceptibility using the Kirby-Bauer Disk Diffusion Method. The antibiotic disc diffusion method showed that S. aureus was resistant to penicillin G, oxytetracycline, sulfamethoxazole, tetracycline, erythromycin, and ampicillin, whereas Salmonella spp. was resistant to penicillin G, sulfamethoxazole, erythromycin, and ampicillin. As these products are consumed frequently, their contamination with S. aureus (≥ 5 × 103 cfu/g) and Salmonella spp. can be a risk factor for food poisoning. The contamination of meat products’ with S. aureus and Salmonella spp. can be a risk factor for public health and the antibiotics to be preferred in illness treatment are of critical importance.

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