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

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.

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Species-specific duplex PCR for the detection of Pratylenchus penetrans

Nematology ◽

10.1163/156854109x428016 ◽

2009 ◽

Vol 11 (6) ◽

pp. 847-857 ◽

Cited By ~ 21

Author(s):

Lieven Waeyenberge ◽

Nicole Viaene ◽

Maurice Moens

Keyword(s):

Internal Control ◽

Dna Sequences ◽

Rrna Gene ◽

Duplex Pcr ◽

Specific Primers ◽

5.8S Rrna Gene ◽

5.8S Rrna ◽

Wide Range ◽

Pcr Products ◽

Species Specific

Abstract ITS1, the 5.8S rRNA gene and ITS2 of the rDNA region were sequenced from 20 different Pratylenchus species. Additionally, the same region was sequenced from seven populations of P. penetrans. After purifying, cloning and sequencing the PCR products, all sequences were aligned in order to find unique sites suitable for the design of species-specific primers for P. penetrans. Since ITS regions showed variability between and even within populations of P. penetrans, only three small DNA sequences were suitable for the construction of three potentially useful species-specific primers. New species-specific primers were paired with existing universal ITS primers and tested in all possible primer combinations. The best performing primer set, supplemented with a universal 28S rDNA primer set that served as an internal control, was tested in duplex PCR. The ideal annealing temperature, Mg2+ concentration and primer ratios were then determined for the most promising primer set. The optimised duplex PCR was subsequently tested on a wide range of different Pratylenchus spp. and 25 P. penetrans populations originating from all over the world. To test the sensitivity, the duplex PCR was conducted on DNA extracted from a single P. penetrans nematode mixed with varying amounts of nematodes belonging to another Pratylenchus species. Results showed that a reliable and sensitive P. penetrans species-specific duplex PCR was constructed.

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Quantitative Detection of Species-Specific DNA in Feedstuffs and Fish Meals

Journal of Food Protection ◽

10.4315/0362-028x-68.6.1217 ◽

2005 ◽

Vol 68 (6) ◽

pp. 1217-1221 ◽

Cited By ~ 27

Author(s):

PAVEL KRCMAR ◽

EVA RENCOVA

Keyword(s):

Mitochondrial Dna ◽

Real Time ◽

Dna Sequences ◽

Real Time Pcr ◽

Single Species ◽

Absolute Quantification ◽

Quantitative Detection ◽

Vertebrate Species ◽

Target Species ◽

Species Specific

A sensitive and rapid method for the quantitative detection of bovine-, ovine-, swine-, and chicken-specific mitochondrial DNA sequences based on real-time PCR has been developed. The specificity of the primers and probes for real-time PCR has been tested using DNA samples of other vertebrate species that may also be present in rendered products. The quantitative detection was performed with dual-labeled probes (TaqMan) using absolute quantification with external standards of single species meat-and-bone meals. This method facilitates the detection of 0.01% of the target species–derived material in concentrate feed mixtures and fish meals.

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Species-specific detection of Candida tropicalis using evolutionary conserved intein DNA sequences

Letters in Applied Microbiology ◽

10.1111/lam.12861 ◽

2018 ◽

Vol 66 (5) ◽

pp. 378-383

Author(s):

S.K. Rajasekharan ◽

A.K. Ray ◽

S. Ramesh ◽

S. Kannappan Mohanvel

Keyword(s):

Dna Sequences ◽

Candida Tropicalis ◽

Specific Detection ◽

Species Specific

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

Journal of Agricultural and Food Chemistry ◽

10.1021/jf9907438 ◽

2000 ◽

Vol 48 (7) ◽

pp. 2829-2832 ◽

Cited By ~ 94

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

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A Novel Pan-FlavivirusDetection and Identification Assay Based on RT-qPCR and Microarray

BioMed Research International ◽

10.1155/2017/4248756 ◽

2017 ◽

Vol 2017 ◽

pp. 1-12 ◽

Cited By ~ 16

Author(s):

Ariel Vina-Rodriguez ◽

Konrad Sachse ◽

Ute Ziegler ◽

Serafeim C. Chaintoutis ◽

Markus Keller ◽

...

Keyword(s):

External Quality Assessment ◽

Virus Detection ◽

Test Results ◽

Rt Pcr ◽

Specific Detection ◽

Field Samples ◽

Pcr Products ◽

Mosquito Pool ◽

Species Specific ◽

Validation Experiments

The genusFlavivirusincludes arthropod-borne viruses responsible for a large number of infections in humans and economically important animals. While RT-PCR protocols for specific detection of mostFlavivirusspecies are available, there has been also a demand for a broad-rangeFlavivirusassay covering all members of the genus. It is particularly challenging to balance specificity at genus level with equal sensitivity towards each target species. In the present study, a novel assay combining a SYBR Green-based RT-qPCR with a low-density DNA microarray has been developed. Validation experiments confirmed that the RT-qPCR exhibited roughly equal sensitivity of detection and quantification for all flaviviruses tested. These PCR products are subjected to hybridization on a microarray carrying 84 different oligonucleotide probes that represent all knownFlavivirusspecies. This assay has been used as a screening and confirmation tool forFlaviviruspresence in laboratory and field samples, and it performed successfully in international External Quality Assessment of NAT studies. Twenty-sixFlavivirusstrains were tested with the assay, showing equivalent or superior characteristics compared with the original or even with species-specific RT-PCRs. As an example, test results on West Nile virus detection in a panel of 340 mosquito pool samples from Greece are presented.

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A Simple and Reliable Single Tube Septuple PCR Assay for Simultaneous Identification of Seven Meat Species

Foods ◽

10.3390/foods10051083 ◽

2021 ◽

Vol 10 (5) ◽

pp. 1083

Author(s):

Zhendong Cai ◽

Song Zhou ◽

Qianqian Liu ◽

Hui Ma ◽

Xinyi Yuan ◽

...

Keyword(s):

Dna Sequences ◽

Animal Species ◽

Meat Product ◽

Meat Products ◽

Pcr Assay ◽

Specific Primers ◽

Meat Species ◽

Pcr Method ◽

Species Specific ◽

Simultaneous Identification

Multiplex PCR methods have been frequently used for authentication of meat product adulteration. Through screening of new species-specific primers designed based on the mitochondrial DNA sequences, a septuple PCR method is ultimately developed and optimized to simultaneously detect seven species including turkey (110 bp), goose (194 bp), pig (254 bp), sheep (329 bp), beef (473 bp), chicken (612 bp) and duck (718 bp) in one reaction. The proposed method has been validated to be specific, sensitive, robust and inexpensive. Taken together, the developed septuple PCR assay is reliable and efficient, not only to authenticate animal species in commercial meat products, but also easily feasible in a general laboratory without special infrastructures.

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Co-structure analysis and genetic associations reveal insights into pinworms (Trypanoxyuris) and primates (Alouatta palliata) microevolutionary dynamics

BMC Ecology and Evolution ◽

10.1186/s12862-021-01924-4 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Brenda Solórzano-García ◽

Ella Vázquez-Domínguez ◽

Gerardo Pérez-Ponce de León ◽

Daniel Piñero

Keyword(s):

Mitochondrial Dna ◽

Host Specificity ◽

Dna Sequences ◽

Evolutionary History ◽

Alouatta Palliata ◽

Howler Monkey ◽

Genetic Associations ◽

Howler Monkeys ◽

Mantled Howler ◽

Species Specific

Abstract Background In parasitism arm race processes and red queen dynamics between host and parasites reciprocally mold many aspects of their genetics and evolution. We performed a parallel assessment of population genetics and demography of two species of pinworms with different degrees of host specificity (Trypanoxyuris multilabiatus, species-specific; and T. minutus, genus-specific) and their host, the mantled howler monkey (Alouatta palliata), based on mitochondrial DNA sequences and microsatellite loci (these only for the host). Given that pinworms and primates have a close co-evolutionary history, covariation in several genetic aspects of their populations is expected. Results Mitochondrial DNA revealed two genetic clusters (West and East) in both pinworm species and howler monkeys, although population structure and genetic differentiation were stronger in the host, while genetic diversity was higher in pinworms than howler populations. Co-divergence tests showed no congruence between host and parasite phylogenies; nonetheless, a significant correlation was found between both pinworms and A. palliata genetic pairwise distances suggesting that the parasites’ gene flow is mediated by the host dispersal. Moreover, the parasite most infective and the host most susceptible haplotypes were also the most frequent, whereas the less divergent haplotypes tended to be either more infective (for pinworms) or more susceptible (for howlers). Finally, a positive correlation was found between pairwise p-distance of host haplotypes and that of their associated pinworm haplotypes. Conclusion The genetic configuration of pinworm populations appears to be molded by their own demography and life history traits in conjunction with the biology and evolutionary history of their hosts, including host genetic variation, social interactions, dispersal and biogeography. Similarity in patterns of genetic structure, differentiation and diversity is higher between howler monkeys and T. multilabiatus in comparison with T. minutus, highlighting the role of host-specificity in coevolving processes. Trypanoxyuris minutus exhibits genetic specificity towards the most frequent host haplotype as well as geographic specificity. Results suggest signals of potential local adaptation in pinworms and further support the notion of correlated evolution between pinworms and their primate hosts.

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Design of primer pairs for species-specific diagnosis of Leishmania (Leishmania) infantum chagasi using PCR

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-29612012000300024 ◽

2012 ◽

Vol 21 (3) ◽

pp. 304-307 ◽

Cited By ~ 4

Author(s):

Osvaldo José da Silveira Neto ◽

Sabrina Castilho Duarte ◽

Hérika Xavier da Costa ◽

Guido Fontgalland Coelho Linhares

Keyword(s):

18S Rrna ◽

18S Rrna Gene ◽

Reference Sequence ◽

Rrna Gene ◽

Analytical Sensitivity ◽

Specific Detection ◽

Pcr Products ◽

Specific Amplification ◽

Pcr Assays ◽

Species Specific

The objective of this study was to design and evaluate new primers for species-specific detection of L. infantum chagasi using PCR. Two combinations of primer pairs were established with the aim of obtaining specific amplification products from the L. infantum chagasi 18S rRNA gene. The combinations of the primer pairs and the respective sizes of the PCR products, based on the U422465 GenBank reference sequence of L. infantum chagasi, were: LCS1/LCS3 (259 bp) and LCS2/LCS3 (820 bp). It was concluded that the new PCR assays optimized using the primer pairs LCS1/LCS3 and LCS2/LCS3 were effective for specific detection of L. infantum chagasi, with analytical sensitivity to detect 1 pg/µL of DNA.

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