Duplex PCR approach for the detection and quantification of donkey, horse and mule in raw and heat-processed meat products

2014 ◽  
Vol 50 (3) ◽  
pp. 834-839 ◽  
Author(s):  
Ailiang Chen ◽  
Chengbin Wei ◽  
Gang Chen ◽  
Yan Zhao ◽  
Shuming Yang
Keyword(s):  
Meat Products ◽  
Processed Meat ◽  
Duplex Pcr ◽  
Detection And Quantification

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

Biomolecules ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 804 ◽  
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.

scholarly journals AUTENTIKASI DAGING AYAM SEGAR DARI KONTAMINASI DAGING BABI MENGGUNAKAN GEN CYT-B DENGAN ANALISIS DUPLEX- POLYMERASE CHAIN REACTION

2017 ◽  
Vol 41 (2) ◽  
pp. 113
Author(s):  
Bayu Setya Hertanto ◽  
Rizky Aulia Fitra ◽  
Lilik Retna Kartikasari ◽  
Muhammad Cahyadi
Keyword(s):  
Meat Products ◽  
Chicken Meat ◽  
Pcr Analysis ◽  
Processed Meat ◽  
Duplex Pcr ◽  
Cyt B ◽  
Polymerase Chain ◽  
Contamination Levels ◽  
The City ◽  
Dna Isolation Protocol

Halal is one of important aspects in consumer protection. Meat and processed meat products are food that should be controlled strictly because those are prone to be adulterated by pork contamination. Therefore, it is necessary to provide detection technique which is accurate, fast and cheap. The objective of this research was to identify the presence of impurities of pork meat on raw chicken meat using gene Cyt-b with duplex-PCR analysis. This research used six samples of raw chicken meat and raw pork. Raw chicken meat was bought from supermarkets in the city of Surakarta and raw pork was obtained from pig slaughterhouse. The percentage of raw pork contamination on raw chicken meat was designed as much as 1, 5, 10, and 25%, respectively. The DNA genome was isolated according to DNA isolation protocol from Genomic DNA Mini Kit. In addition, duplex-PCR was performed based on protocol of KAPA2G Fast Multiplex PCR kit. The data was descriptively analyzed by directly looking the DNA bands on the gel documentation apparatus. The result showed that specific DNA bands for chicken and pig were completely appeared on 1.5% of agarose gels. Duplex-PCR detect contamination of pork on raw meat of chicken at all contamination levels. This research proved that the duplex-PCR detect the contamination of pork until the level of 1%.

scholarly journals Low cost and comprehensive pork detection in processed food products with a different food matrix

2018 ◽  
Vol 23 (1) ◽  
pp. 21
Author(s):  
Fenny Aulia Sugiana ◽  
Henni Widyowati ◽  
Muhammad Ali Warisman ◽  
Suryani Suryani ◽  
Desriani Desriani
Keyword(s):  
18S Rrna ◽  
Low Cost ◽  
Meat Products ◽  
Positive Control ◽  
Negative Control ◽  
Dna Integrity ◽  
Processed Meat ◽  
Duplex Pcr ◽  
Processed Food ◽  
Food Matrix

The adulteration of processed beef-based meat products with pork is a sensitive issue in Indonesia. In this study, we developed a detection method for the low cost identification of pork in processed meat products. We used the cost-efficient Taq DNA polymerase, DreamTaq Green PCR master mix (2x), and duplex PCR method to recognize pork simultaneously with 18S rRNA detection. A positive control containing a pork gene inserted into pGEM®-T easy was prepared, along with a negative control. The results of the duplex PCR were used to assess its specificity, detection limit, and its ability to recognize pork in processed meat products with a different food matrix. 18S rRNA detection was for confirming DNA integrity of DNA extracted from the processed food, while the positive control confirmed that the reagents were working well and the negative control confirmed a non-contamination problem. Following this, the duplex PCR was optimized and the optimum concentration primer for duplex PCR detection was found to be 3 µm for pork and 0.2 µm for 18S rRNA. As little as 3.125 ng of the DNA template could be used to detect whether a sample contained pork. Duplex PCR is a simple, fast, sensitive, specific, and low cost method of detecting pork in processed meat products.

scholarly journals An Ultrasound Assessed Extraction Combined with Ion-Pair HPLC Method and Risk Assessment of Nitrite and Nitrate in Cured Meat

2018 ◽  
Vol 2018 ◽  
pp. 1-6 ◽  
Author(s):  
Babiker Yagoub Abdulkair ◽  
Amin O. Elzupir ◽  
Abdulaziz S. Alamer
Keyword(s):  
Risk Assessment ◽  
Tetrabutylammonium Bromide ◽  
Meat Products ◽  
Correlation Coefficients ◽  
Dietary Exposure ◽  
Hplc Method ◽  
Processed Meat ◽  
Disodium Hydrogen Phosphate ◽  
Cured Meat

An accurate IPC-UV method was developed and validated for the determination of nitrite (NI) and nitrate (NA) in meat products. The best separation was achieved on a phenyl-hexyl column (150 mm × 4.6 mm, 3 µm) with a mobile phase composed of 25% acetonitrile and 75% buffer (2 mM disodium hydrogen phosphate and 3 mM tetrabutylammonium bromide, pH = 4). Eluents were monitored at 205 nm. Linearity ranges were 1.86 × 10−6–7.5 µg·ml−1 and 0.09–5.0 µg·ml−1 for NI and NA, respectively. The correlation coefficients were greater than 0.999 for NI and NA. This method was applied to a number of processed meat products in Riyadh (n = 155). NI ranged from 1.78 to 129.69 mg·kg−1, and NA ranged from 0.76 to 96.64 mg·kg−1. Results showed extensive use of NI and NA; however, concentrations were within the legal limit of Saudi Arabia except for one sample. Further, the risk assessment and dietary exposure have been estimated for both NI and NA.

Occurrence and Survival of Verocytotoxin-Producing Escherichia coli O157 in Meats Obtained from Retail Outlets in The Netherlands

1999 ◽  
Vol 62 (10) ◽  
pp. 1115-1122 ◽  
Author(s):  
A. E. HEUVELINK ◽  
J. T. M. ZWARTKRUIS-NAHUIS ◽  
R. R. BEUMER ◽  
D E. de BOER
Keyword(s):  
Escherichia Coli ◽  
The Netherlands ◽  
Meat Products ◽  
Storage Period ◽  
Processed Meat ◽  
Raw Meat ◽  
Pork Products ◽  
Minced Beef ◽  
Beef Products ◽  
Meat Samples

In 1996 and 1997, 2,941 fresh and processed meat products obtained from supermarkets and butcher shops in The Netherlands were examined for the presence of verocytotoxin-producing Escherichia coli of serogroup O157 (O157 VTEC). Additionally, the fate of O157 VTEC in raw meat products stored at low temperatures and the effect of different additives were evaluated. O157 VTEC strains were isolated from 6 (1.1%) of 571 samples of raw minced beef, 2 (0.5%) of 402 samples of raw minced mixed beef and pork, 1 (1.3%) of 76 samples of raw minced pork, 1 (0.3%) of 393 samples of other raw pork products, and 1 (0.3%) of 328 samples of cooked or fermented ready-to-eat meats. Other raw beef products (n = 223) and meat samples originating from poultry (n = 819), sheep or lamb (n = 46), or wild animals (n = 83) were all found to be negative for O157 VTEC. For the survival experiments we used tartaar (minced beef with a fat content of less than 10%) and filet americain (tartaar mixed with a mayonnaise-based sauce [80 to 20%]). The O157 VTEC strain tested was able to survive in tartaar and filet americain stored at −20, 0, 5, or 7°C for 3 days. At both 7 and at 15°C, O157 VTEC counts in tartaar and filet americain remained virtually unchanged throughout a storage period of 5 days. Addition of acetic acid (to pH 4.0), sodium lactate (1 and 2% [wt/wt]), or components of the lactoperoxidase–thiocyanate–hydrogen peroxide system to filet americain did not result in a reduction of viable O157 VTEC cells during storage at 7 or 15°C. It was concluded that raw meat contaminated with O157 VTEC will remain a hazard even if the meat is held at low or freezing temperatures.

scholarly journals Molecular and HPLC-based approaches for detection of aflatoxin B1 and ochratoxin A released from toxigenic Aspergillus species in processed meat

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Abdelazeem M. Algammal ◽  
Mahmoud E. Elsayed ◽  
Hany R. Hashem ◽  
Hazem Ramadan ◽  
Norhan S. Sheraba ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Meat Products ◽  
Its Region ◽  
Processed Meat ◽  
Isolation And Identification ◽  
Beef Meat ◽  
Meat Samples ◽  
Aflatoxin B ◽  
Minced Meat ◽  
Mold Count

Abstract Background Meat-products are considered an enriched media for mycotoxins. This study aimed to investigate the prevalence of toxigenic Aspergillus species in processed meat samples, HPLC-quantitative measurement of aflatoxin B1 and ochratoxin A residues, and molecular sequencing of aflR1 and pks genes. One hundred and twenty processed beef meat specimens (basterma, sausage, and minced meat; n = 40 for each) were collected from Ismailia Province, Egypt. Samples were prepared for total mold count, isolation, and identification of Aspergillus species. All samples were analyzed for the production of both Aflatoxin B1 and Ochratoxin A mycotoxins by HPLC. Molecular identification of Aspergillus flavus and Aspergillus ochraceus was performed using PCR amplification of the internal transcribed spacer (ITS) region; furthermore, the aflR1 and pks genes were sequenced. Results The total mold count obtained from sausage samples was the highest one, followed by minced meat samples. The prevalence of A. flavus was (15%), (7.5%), and (10%), while the prevalence of A. ochraceus was (2.5%), (10%), and (0%) in the examined basterma, sausage, and minced meat samples, respectively. Using PCR, the ITS region was successfully amplified in all the tested A. flavus and A. ochraceus strains. Aflatoxin B1 was detected in six basterma samples (15%). Moreover, the ochratoxin A was detected only in four sausage samples (10%). The aflR1 and pks genes were amplified and sequenced successfully and deposited in the GenBank with accession numbers MF694264 and MF694264, respectively. Conclusions To the best of our knowledge, this is the first report concerning the HPLC-Molecular-based approaches for the detection of aflatoxin B1 and ochratoxin A in processed beef meat in Egypt. The production of aflatoxin B1 and ochratoxin A in processed meat constitutes a public health threat. Aflatoxin B1 is commonly associated with basterma samples. Moreover, ochratoxin A was detected frequently in sausage samples. The routine inspection of mycotoxins in processed meat products is essential to protect human consumers.

DEVELOPMENT OF IMMUNOENZYME ANALYTICAL SYSTEMS FOR CONTROL OF THE CONTENT OF MAIN NON-MEAT COMPOUNDS IN PROCESSED MEAT PRODUCTS

2021 ◽  
Vol 1 (19) ◽  
pp. 322-324
Author(s):  
E.A. Zvereva ◽  
A.V. Zherdev ◽  
B.B. Dzantiev
Keyword(s):  
Connective Tissue ◽  
Enzyme Immunoassay ◽  
Trypsin Inhibitor ◽  
Meat Products ◽  
Soybean Trypsin Inhibitor ◽  
Processed Meat

Methods have been developed to control the content of non-meat components (connective tissue of animals, eggs, soy) in processed meat products, based on enzyme immunoassay of biomarker proteins – collagen, ovalbumin, soybean trypsin inhibitor.

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