Development of a real-time PCR assay for the detection of Trichinella spiralis in muscle tissue of swine and derivatives.

SILVINA QUINTANA; MARIANA RECAVARREN; EXEQUIEL SCIALFA; VIERA I; M. RIVERO; KRIVOKAPICH M

JOURNAL OF FOOD SAFETY

WILEY-BLACKWELL PUBLISHING, INC

Año: 2015 vol. 36 p. 282 - 287

0149-6085

The purpose of this study was to generate a preventive tool for a sensitive and specific molecular diagnostic for trichinellosis, that represent one of the most important zoonotic diseases in Argentina. A real-time PCR method was developed to detect Trichinella spiralis DNA and was used to analyze 21 muscles samples of swine muscles and its derivatives suspected of trichinellosis. Extraction, amplification and quantification of DNA from muscle samples were performed by real-time PCR and. , Iin order to check the success of DNA extraction and the lack of inhibition, an internal porcine DNA amplification control was included. The developed PCR assay specifically detects T. spiralis showing no amplification with other Trichinella genotypes and showed a sensitivity of 0,024 LPG in swine muscle samples. Five samples negative by enzymatic digestion were positive by PCR, which demonstrates that this technique is more sensitive than gold standard technique. Positive results indicate a higher risk of infection for outdoor farming in areas where Trichinella is endemic. This is the first real-time PCR assay for T. spiralis detection with internal control.