Direct and Highly Species-Specific Detection of Pork Meat and Fat in Meat Products by PCR Amplification of Mitochondrial DNA

2000 ◽  
Vol 48 (7) ◽  
pp. 2829-2832 ◽  
Author(s):  
J. F. Montiel-Sosa ◽  
E. Ruiz-Pesini ◽  
J. Montoya ◽  
P. Roncalés ◽  
M. J. López-Pérez ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Meat Products ◽  
Pcr Amplification ◽  
Pork Meat ◽  
Specific Detection ◽  
Species Specific

Cloth-Based Hybridization Array System for Expanded Identification of the Animal Species Origin of Derived Materials in Feeds

2007 ◽  
Vol 70 (12) ◽  
pp. 2900-2905 ◽  
Author(s):  
JOHANNA MURPHY ◽  
JENNIFER ARMOUR ◽  
BURTON W. BLAIS
Keyword(s):  
Mitochondrial Dna ◽  
Dna Sequences ◽  
Microscopic Examination ◽  
Animal Species ◽  
Specific Detection ◽  
Animal Feeds ◽  
Species Origin ◽  
Pcr Products ◽  
Species Specific ◽  
Immunoenzymatic Assay

A cloth-based hybridization array system (CHAS) previously developed for the detection of animal species for which prohibited materials have been specified (cattle, sheep, goat, elk, and deer) has been expanded to include the detection of animal species for which there are no prohibitions (pig and horse) in Canadian and American animal feeds. Animal species were identified by amplification of mitochondrial DNA sequences by PCR and subsequent hybridization of the amplicons with an array of species-specific oligonucleotide capture probes immobilized on a polyester cloth support, followed by an immunoenzymatic assay of the bound PCR products. The CHAS permitted sensitive and specific detection of meat meals from different animal species blended in a grain-based feed and should provide a useful adjunct to microscopic examination for the identification of prohibited materials in animal feeds.

Defeating numts: Semi-pure mitochondrial DNA from eggs and simple purification methods for field-collected wildlife tissues

Genome ◽  
2006 ◽  
Vol 49 (11) ◽  
pp. 1438-1450 ◽  
Author(s):  
Gabriela Ibarguchi ◽  
Vicki L. Friesen ◽  
Stephen C. Lougheed
Keyword(s):  
Mitochondrial Dna ◽  
Gradient Methods ◽  
Pcr Amplification ◽  
Cesium Chloride ◽  
Pcr Cloning ◽  
Degenerate Primers ◽  
Purification Methods ◽  
Mitochondrial Origin ◽  
Reference Sequences ◽  
Species Specific

Mitochondrial DNA (mtDNA) continues to play a pivotal role in phylogeographic, phylogenetic, and population genetic studies. PCR amplification with mitochondrial primers often yields ambiguous sequences, in part because of the coamplification of nuclear copies of mitochondrial genes (numts) and true mitochondrial heteroplasmy arising from mutations, hybridization with paternal leakage, gene duplications, and recombination. Failing to detect numts or to distinguish the origin of such homologous sequences results in the incorrect interpretation of data. However, few studies obtain purified mtDNA to confirm the mitochondrial origin of the first reference sequences for a species. Here, we demonstrate the importance and ease of obtaining semi-pure mtDNA from wildlife tissues, preserved under various typical field conditions, and investigate the success of 3 commercial extraction kits, cesium-chloride gradient mtDNA purification, long-template PCR amplification, cloning, and more species-specific degenerate primers. Using more detailed avian examples, we illustrate that unfertilized or undeveloped eggs provide the purest sources of mtDNA; that kits provide an alternative to cesium-chloride gradient methods; and that long-template PCR, cloning, and degenerate primers cannot be used to produce reliable mitochondrial reference sequences, but can be powerful tools when used in conjunction with purified mtDNA stocks to distinguish numts from true heteroplasmy.

Application of mitochondrial DNA analysis for microbial source tracking purposes in shellfish harvesting waters

2010 ◽  
Vol 61 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Craig Baker-Austin ◽  
Rachel Rangdale ◽  
James Lowther ◽  
David N. Lees
Keyword(s):  
Mitochondrial Dna ◽  
Real Time ◽  
Real Time Pcr ◽  
Dna Analysis ◽  
Pcr Amplification ◽  
Source Tracking ◽  
Pcr Analysis ◽  
Taqman Probes ◽  
Faecal Matter ◽  
Species Specific

We present a method for the reliable detection and source characterisation of faecal pollution in water and shellfish matrices, utilising real-time PCR analysis of mitochondrial DNA targets. In this study we designed real-time PCR (TaqMan) probes to target human, bovine, ovine and swine mtDNA. PCR amplification using species-specific TaqMan probes on faecal matter and mixed effluent slurries revealed no cross-reactions between species of interest and other vertebrate faecal matter. Performed as a single blind experiment we were able to correctly identify faecal material in 17/20 effluents (85% correct). mtDNA degrades relatively quickly in faecally-spiked water samples (∼2 weeks), a similar timeframe of environmental persistence to several bacterial faecal indictors, highlighting its applicability. The procedure described here is specific, rapid (<5 hours) and sensitive. These results confirm the suitability of using species-specific mtDNA as an indicator in source tracking studies in surface waters, shellfish harvesting areas and shellfish matrices.

Trypanosoma vivax: Characterization of the Spliced-Leader Gene of a Brazilian Stock and Species-Specific Detection by PCR Amplification of an Intergenic Spacer Sequence

2001 ◽  
Vol 99 (1) ◽  
pp. 37-48 ◽  
Author(s):  
Rogéria M Ventura ◽  
Fernando Paiva ◽  
Roberto A.M.S Silva ◽  
Gentilda F Takeda ◽  
Gregory A Buck ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Intergenic Spacer ◽  
Trypanosoma Vivax ◽  
Specific Detection ◽  
Spliced Leader ◽  
Species Specific

Species-specific Detection of Campylobacters Important in Veterinary Medicine by PCR Amplification of 23S rDNA Areas

1995 ◽  
Vol 17 (4) ◽  
pp. 563-568 ◽  
Author(s):  
Katrin Bastyns ◽  
Sabine Chapelle ◽  
Peter Vandamme ◽  
Herman Goossens ◽  
Rupert De Wachter
Keyword(s):  
Veterinary Medicine ◽  
Pcr Amplification ◽  
Specific Detection ◽  
Species Specific

scholarly journals Listeriosis in a peri-urban area: Cultural and molecular characterization of Listeria monocytogenes isolated from encephalitic goats

2020 ◽  
Vol 13 (9) ◽  
pp. 1743-1749
Author(s):  
Nagendra Nath Barman ◽  
Anjan Jyoti Nath ◽  
Sharmita Doley ◽  
Shameem Ara Begum ◽  
Parikshit Kakati ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
16S Rrna ◽  
Molecular Characterization ◽  
Early Stage ◽  
Meat Products ◽  
Pcr Amplification ◽  
High Dose ◽  
Molecular Tests ◽  
Species Specific

Background and Aim: Listeriosis in food animals bears a significant threat to human health. Detailed investigations into the cause facilitate proper management of the disease. This study reports the cultural, pathological, and molecular characterization of Listeria monocytogenes isolated from encephalitic goats from peri-urban Guwahati, Assam. Materials and Methods: Out of nine suspected samples, five positive isolates of L. monocytogenes were subjected to bacteriological, biochemical, and molecular tests. The genus and species-specific L. monocytogenes 16S rRNA and prs genes were amplified by polymerase chain reaction (PCR) to yield 1200 and 370 bp sized products, respectively. The encephalitic form of the disease was characterized by circling movement, high fever, and terminal recumbence. Results: All the five isolates were confirmed to be L. monocytogenes based on PCR amplification of genus and species-specific 16S rRNA and prs gene products. The isolates were sensitive to ciprofloxacin, oxytetracycline (OTC), and norfloxacin, but resistant to doxycycline and erythromycin. A high dose of OTC was used in a goat at the early stage of clinical symptom and the animal recovered clinically. Conclusion: Listeriosis in goats could pose a significant public health threat as the meat (occasionally milk) or meat products from goats are widely consumed by the people of Assam. Understanding the molecular epidemiological aspects of L. monocytogenes infections of food animal species should, therefore, be the priority in this part of the country.

64: Rapid, Species-Specific Detection of Uropathogens from Clinical Urine Specimens Using an Electrochemical Microsensor Array

2005 ◽  
Vol 173 (4S) ◽  
pp. 18-18
Author(s):  
Joseph C. Liao ◽  
Mitra Mastali ◽  
David A. Haake ◽  
Bernard M. Churchill
Keyword(s):  
Specific Detection ◽  
Species Specific ◽  
Urine Specimens

scholarly journals Multispecies Identification of Oilseed- and Meat-Specific Proteins and Heat-Stable Peptide Markers in Food Products

Molecules ◽  
2021 ◽  
Vol 26 (6) ◽  
pp. 1577
Author(s):  
Klaudia Kotecka-Majchrzak ◽  
Natalia Kasałka-Czarna ◽  
Agata Sumara ◽  
Emilia Fornal ◽  
Magdalena Montowska
Keyword(s):  
Limit Of Detection ◽  
Raw Materials ◽  
Meat Products ◽  
Food Products ◽  
Plant Raw Materials ◽  
Heat Stable ◽  
2S Albumins ◽  
Specific Proteins ◽  
Meat Species ◽  
Species Specific

Consumer demand for both plant products and meat products enriched with plant raw materials is constantly increasing. Therefore, new versatile and reliable methods are needed to find and combat fraudulent practices in processed foods. The objective of this study was to identify oilseed species-specific peptide markers and meat-specific markers that were resistant to processing, for multispecies authentication of different meat and vegan food products using the proteomic LC-MS/MS method. To assess the limit of detection (LOD) for hemp proteins, cooked meatballs consisting of three meat species and hemp cake at a final concentration of up to 7.4% were examined. Hemp addition at a low concentration of below 1% was detected. The LOD for edestin subunits and albumin was 0.9% (w/w), whereas for 7S vicilin-like protein it was 4.2% (w/w). Specific heat-stable peptides unique to hemp seeds, flaxseed, nigella, pumpkin, sesame, and sunflower seeds, as well as guinea fowl, rabbit, pork, and chicken meat, were detected in different meat and vegan foods. Most of the oilseed-specific peptides were identified as processing-resistant markers belonging to 11S globulin subunits, namely conlinin, edestin, helianthinin, pumpkin vicilin-like or late embryogenesis proteins, and sesame legumin-like as well as 2S albumins and oleosin isoforms or selected enzymic proteins.

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