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On point identification of species origin of food animals by recombinase polymerase amplification-lateral flow (RPA-LF) assay targeting mitochondrial gene sequences.
Kumar, Dhananjay; Kumar, Rajiv Ranjan; Rana, Preeti; Mendiratta, S K; Agarwal, R K; Singh, Praveen; Kumari, Sarita; Jawla, Jyoti.
Afiliación
  • Kumar D; Division of Livestock Products Technology, Indian Veterinary Research Institute, Izatnagar, Bareilly, 243122 India.
  • Kumar RR; Division of Livestock Products Technology, Indian Veterinary Research Institute, Izatnagar, Bareilly, 243122 India.
  • Rana P; Department of Livestock Products Technology, CVASc, DUVASU, Mathura, India.
  • Mendiratta SK; Division of Livestock Products Technology, Indian Veterinary Research Institute, Izatnagar, Bareilly, 243122 India.
  • Agarwal RK; Division of Livestock Products Technology, Indian Veterinary Research Institute, Izatnagar, Bareilly, 243122 India.
  • Singh P; Division of Vet Biotechnology, Indian Veterinary Research Institute, Izatnagar, Bareilly, India.
  • Kumari S; Department of Livestock Products Technology, PGIVER, RAJUVAS, Jaipur, India.
  • Jawla J; Division of Livestock Products Technology, GADVASU, Ludhiana, India.
J Food Sci Technol ; 58(4): 1286-1294, 2021 Apr.
Article en En | MEDLINE | ID: mdl-33746256
The present study was aimed to develop and standardize Recombinase polymerase amplification-lateral flow (RPA-LF) assays for on point identification of species origin of food animals viz: cattle, buffalo and pig. Species specific RPA primers sets for cattle, buffalo and pig were designed by homology comparisons of the sequences of mitochondrial cytochrome b gene and d-loop region from common food species viz: cattle, buffalo, sheep, goat, pig and chicken. The RPA assays for designed primers sets were optimized using the reaction components from Twist Amp basic kit and instructions in its manual. Endpoint detection of species specific amplified RPA products were made by gel electrophoresis and designed species specific RPA-LFA strips. The developed assays were evaluated for their specificity, diagnostic sensitivity, and validated on coded samples and binary meat admixtures with relative percentage of 20, 10, 5 & 1% target species. The developed RPA assays resulted in amplification of DNA template exclusively of cattle, buffalo and pig origin to product sizes of 294, 405 and 283 bp respectively. The diagnostic sensitivities of developed assays were up to 10 pg of genomic DNA and highly correlated with species specific PCR assays taken as gold standard. Developed species specific RPA assays also identified the target species in coded samples and binary meat admixture up to 1%.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Tipo de estudio: Diagnostic_studies / Guideline Idioma: En Revista: J Food Sci Technol Año: 2021 Tipo del documento: Article Pais de publicación: India

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Tipo de estudio: Diagnostic_studies / Guideline Idioma: En Revista: J Food Sci Technol Año: 2021 Tipo del documento: Article Pais de publicación: India