meat species identification (23)

 

10886144077?profile=RESIZE_584xIn this paper, which also utilised CRISPR, the researchers combined loop-mediated isothermal amplification (LAMP) with a sensitive CRISPR-Cas12a silver nanocluster fluorescence sensing system, for testing for adulteration in meat samples. The specific sequence recognition of CRISPR-Cas12a allowed accurate identification of the target DNA. 

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 10861696675?profile=RESIZE_400xThe sale of halal foods has increased considerably making it a priority to develop methods to authenticate halal certification. This review exams the development of rapid methods based on DNA analysis, which principally determine meat species.  In this review, several rapid DNA techniques such as multiplex PCR, convection PCR, PCR-RFLP, PCRstrip, real-time PCR, LAMP, nanotechnology, and commercial rapid test kits for the detection of porcine DNA and DNA from other animals for halal verification are discussed.

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This study aimed to construct a two-tube hexaplex PCR (polymerase chain reaction) assay to authenticate twelve meat species. Primers were specifically designed to identify the target 12 species - horse, pigeon, camel, rabbit, ostrich, beef, as well as turkey, dog, chicken, duck, cat, and goose, and no untargeted primers were used. The assay was found to be specific, sensitive and reproducible. The assay was also tested on DNA extracted from meat which had been boiled or  microwave-cooked, and was still able to identify the meat species. The proof of principle to  identify many species simultaneously has been shown in this study, and could be adapted to cover different or less/more species.

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Russian scientists have developed a rapid test for chicken and pork in processed meat products, which takes 33 minutes to complete and can be done in-situ without the need of laboratory equipment. The test consists of a 3 min of crude DNA extraction, 20 min of recombinase polymerase amplification (RPA) at 39 °C, and 10 min of lateral flow assay (LFA) detection. The RPA–LFA assay is based on designed fluorescein and biotin-labelled primers developed from the cytochrome B gene, which can identify as little as 0.001% w/w  of the target meat component. The assay was tested on  processed meat products and to meat after heating, and the results were confirmed by real-time PCR.

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Meat adulteration/substitution has long been a source of concern for a variety of reasons, including public health, religious considerations, wholesomeness, unhealthy competitiveness in the meat industry, and fraud. Therefore, a range of kits have been developed to permit rapid, precise, and specific identification of meat species. These are based on DNA methods (PCR and Real-Time PCR) and Enzyme-Linked Immunosorbent Assay (ELISA). In response to these developments, Advance Market Analytics has published a new research publication on “Global Meat Species Identification Kits Market Insights, to 2026”.

The report has 9 chapters:

Chapter 1: Introduction, market driving force product Objective of Study and Research Scope the Meat Species Identification Kits market

Chapter 2: Exclusive Summary – the basic information of the Meat Species Identification Kits Market.

Chapter 3: Displaying the Market Dynamics- Drivers, Trends and Challenges & Opportunities of the Meat Species Identification Kits

Chapter 4: Presenting the Meat Species Identification Kits Market Factor Analysis, Porters Five Forces, Supply/Value Chain, PESTEL analysis, Market Entropy, Patent/Trademark Analysis.

Chapter 5: Displaying the by Type, End User and Region/Country 2015-2020

Chapter 6: Evaluating the leading manufacturers of the Meat Species Identification Kits market which consists of its Competitive Landscape, Peer Group Analysis, BCG Matrix & Company Profile

Chapter 7: To evaluate the market by segments, by countries and by Manufacturers/Company with revenue share and sales by key countries in these various regions (2021-2026)

Chapter 8 & 9: Displaying the Appendix, Methodology and Data Source

 A free example copy of the report can be obtained.

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This assay was developed to have a cheaper, simpler and more rapid assay to detect meat species substitution. A novel direct lysis (DL) method was used to extract DNA directly from meat tissue, and to obtain a sample of DNA for multiplex PCR within 15 min, which is more rapid than using a commercial lysis kit.  Four pairs of high-specificity primers for the mitochondrial D-loop region of beef, pork, chicken and duck were designed. When optimised, the assay could detect pork, chicken and duck down to 0.1% (w/w) in meat mixtures even when the meat sample had undergone freezing, heating and autoclaving. The assay was evaluated by testing 79 commercial beef products collected in local markets, and found that nearly 28% of these contained pork or chicken to varying degrees. The accuracy of the results was verified by repeating the analysis using a standard real-time PCR. 

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Malayasian researchers have developed an assay to simultaneously determine 7 meat species (beef, buffalo, chicken, duck, sheep, goat and pork) in processed meat products. Species specific primers to the 7 species were designed, which target the mitochondrial cytochrome b (cytb) and NADH dehydrogenase subunit 5 (ND5) genes, to amplify short DNA fragments (73-263 bp) by PCR (polymerase chain reaction). These were then treated with 3 restriction enzymes ( FatI, BfaI, and HPY188I) to cut the amplicons down into smaller fragments, which were separated by gel electrophoresis. The bp (base pair) length and number of these fragments are unique to each of the species. The assay was tested against 25 non-target species to ensure specificity to the 7 target species, and the limit of detection was determined as 0.5% (w/w) in different matrices. The assay worked on heat treated meat products. A survey of local market meat products detected  buffalo DNA in 84% of commercial beef burgers and frankfurter products tested.

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6733964462?profile=RESIZE_400xThis project aimed to develop and validate a multi-species proteomics screening tool for meat species verification in processed meat products. The method covers nine meat species: beef, pork, horse, goat, lamb, donkey, rabbit, chicken and turkey. The research has improved the original database of marker peptides for the nine animal species studied. The work has expanded previous research using high-resolution mass spectrometry to achieve detection of pork, horse, donkey, lamb, rabbit and chicken at 1% (w/w) adulteration levels. It has developed and validated a triple quadrupole mass spectrometry method suitable for simultaneous identification of the nine meat species in processed meat products. Beef, pork, horse and chicken products were used to carry out an intra-laboratory method validation.

Project FA0166 is in the Research Section of the website

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5766949475?profile=RESIZE_400xPolish researchers have used an LC-QTOF-MS/MS (Liquid Chromatography - Quadrupole Time of Flight - Mass Spectrometer) approach for detecting and identifying rabbit-specific peptide-markers from thermally processed meat products to differentiate rabbit from other commonly-consumed animal species. The instrument identified 49 heat stable peptide markers from rabbit myofibrillar and sarcoplasmic proteins. When 11 heat treated rabbit based pâtés were analysed, 3 of the 49 heat-stable peptides were consistently detected in all the pâté samples and hence considered robust markers for rabbit. Pork, lamb and chicken-specific peptides were also monitored in the pâté samples, and undeclared chicken was found in two of the pâtés.

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4439874075?profile=RESIZE_400xA novel method to identify animal species in complex or adulterated processed meat products has been developed by Chinese researchers, which combines a cytochrome oxidase I (COI) mini-barcode with next-generation sequencing (NGS).  A universal primer based on 140 sequences from 51 edible animal species was designed. A mixture of 12 species raw meat samples (beef, water buffalo, pork, sheep, chicken, partridge, grass carp, silver carp, blue scad, tile fish, pomfret, and prawn) were identified both with the clone sequencing and also with the mini-barcode (136 bp) sequence combined NGS method. The NGS method was superior in accuracy, sensitivity, and detection efficiency compared to the clone sequencing method, The edible animal species were identified both in the mixed raw samples and 7 heavily processed food products (different meatballs - beef, pork, fish, and shrimp), 'modified' beefsteak, sausages, and Chinese sausages. Moreover, some unlabelled species and dubious contamination were also detected as well, leading to stringent cleaning procedures.

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3887798641?profile=RESIZE_710xThere is a need for accurate methods to quantify meat species in meat products to check QUID (quantitive ingredient delarations). Chinese researchers have evaluated a real-time PCR method based on species-specific primers and probes from the mitochondrial cytb (cytochrome b) gene fragment for the identification and quantification of beef ingredient in commercial meat products. The method was developed and calibrated using three mixed matrices (pork, donkey and sheep with known proportions of beef, respectively). Results showed that the primers and probes were highly specific for beef in meat products, and the absolute detection limit of the real-time PCR method was 0.025 ng DNA, and the relative detection limit was 0.002% (w/w) of beef.  The assay was validated with 22 commercial beef products, of which 11 were salted, 10 were jerkies (dried) and one meatball, which were collected from local supermarkets. The results indicated the assay had a good stability in detecting and quantifying beef in the commercial samples.  

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This paper reports the performance of a commercial NGS method that has been evaluated as an untargeted tool to identify meat species. The method was tested on pure meat samples, and all species were correctly identified including several exotic species. Closely related meat species were also correctly differentiated. Species were successfully detected and identified in mixtures down to 1% (w/w). The reliability of the method was further confirmed on several proficiency test samples, and promising quantification data were obtained. Finally, 45 minced meat samples sourced from local European and Asian markets were analysed, and 18% of them showed cases of adulteration with undeclared meat species.

3722359646?profile=RESIZE_710x Read the abstract here

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Chinese researchers have developed a multiplex PCR assay using universal primers based on mitochondrial DNA to identify 8 species (dog, chicken, cattle, pig, horse, donkey, fox, and rabbit) simultaneously in meat products. The assay was tested on 103 commercial meat products from the Chinese market, which demonstrated its effectiveness and applicability.

3571056540?profile=RESIZE_710x Read the abstract here

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Chinese researchers have developed a method for the rapid visual detection of adulterated meat based on both the lateral flow strip (LFS) platform and on polymerase chain reaction (PCR). After the rapid extraction of genomic components from meat, the on-site amplification of the target DNA of  duck meat is carried out with the rationally designed functional FITC- and biotin-modified primer set, thereby producing numerous double-stranded DNA  products dually labelled with FITC and biotin. The FITC-labelled terminal end of the products binds to the pre-immobilized FITC antibody on the test line of the strip, and the biotin-labelled terminal end binds to the streptavidin-conjugated gold nanoparticles, resulting in a visible test line on the LFS for the rapid identification of duck meat in adulterated beef. After optimization, an adulteration ratio as low as 0.05% can be easily measured. Twenty two commercial processed meat samples were tested with this new method, and 4 adulterated samples were successfully identified by both the routine PCR method and the new LFS method. The LFS method is simple in design, convenient in operation, and can be easily extended to the identification of other adulterating meat species just by replacing the modified primers. 

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Example of a lateral flow strip for target DNA detection

 

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Researchers have a developed a multiplex PCR (polymerase chain reaction) assay to identify beef, pork, horse and poultry (chicken, turkey) and determine these species quantitatively in meat products. The qualitative assay uses a mitochondrial cytochrome b gene marker. The quantitative assay uses singlecopy markers from chromosomal genes (cyclic-GMP-phosphodiesterase gene for cattle, beta-actin gene for pig, interleukin-2 gene for chicken), and the normaliser is from the myostatin gene for mammals and poultry. The reliability of both methods was confirmed by analysing of mixed samples prepared with or without heat treatment. The assay was tested with 14 meat products from the Czech retail market with two having undeclared species and another 4 products giving an incorrect quantitative declaration. 

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Conventional nucleic acid amplification methods are reliable, but the requirement of complex equipment, skilled technicians and long operation time limit their on-site use. Here, a simple denaturation bubble-mediated strand exchange amplification method (SEA) requiring only a pair of primers and one polymerase was first reported for identifying adulteration of pork source by targeting the species-specific mitochondrial DNA sequence. The SEA method could detect 1% pork total DNA by both colorimetric and fluorescence determination. The whole detection process could be finished within 1 hour by coupling with fast tissue DNA extraction method, only requiring a simple heating block, and hence making it suitable for on-site meat species identification. 

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In this study, NGS (next generation sequencing) using the Ion Torrent semiconductor platform was applied to identify meat species in several highly processed and complex meat products and meat derived broths (a döner kebab, a beef/pork paté, a meat based filling of tortellini, one instant granular preparation of broth stock made from meat, and two ready-to-use meat broths from different producers). Sequence analysis of reads from 6 libraries detected expected and unexpected meat species in the products. A measure of poor hygienic practice during production of the analysed products could be inferred using the number of human reads. In conclusion, NGS data is useful for authentication of highly processed products.

Read the abstract at: NGS of meat products

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New Book on Proteomics in Food Science Published.

Academic Press has published a  book - Proteomics in Food Science: From Farm to Fork, which is  a useful reference, providing concepts and practical applications of proteomics for in various disciplines of food science. The book covers a range of methods for elucidating the identity or composition of specific proteins in foods or cells related to food science, from spoilage organisms, to edible components. Of interest to food authenticity, there are chapters on soya proteomics, meat species identification, and fish and other seafood identification.

Read the abstract at: food proteomics or a preview at: proteomics in food science

 

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This study evaluated the application of relative quantification of unique heat-stable species specific peptides in highly processed meat proteins. Using nano-LC-QTOF-MS/MS, 20 new, heat-stable peptide markers unique to chicken, duck and goose were identified. The method enabled detection of 1% (w/w) of chicken and 1% (w/w) pork in a mixture of the meat of three species, as well as 0.8% (w/w) beef proteins in commercial poultry frankfurters. This method includes a correction factor for each protein, based on the peptide MS detection probabilities, which are influenced by the physicochemical properties of the peptide. Considerable differences in abundance of myofibrillar and sarcoplasmic proteins were observed between samples and illegal proportions of ingredients were discovered.

Read the abstract at: Meat species quantification using peptide markers

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In this study, rapid identification of meat species was achieved using a portable real-time PCR system, following a very simple DNA extraction method. Applying these techniques, beef, pork, chicken, rabbit, horse, and mutton were correctly identified in processed foods in 20 min. This approach is expected to significantly contribute to factory quality control and fraud mitigation.

Read the abstract at: Portable RT-PCR meat species identification

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