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

Abstract

Trichinellosis is an emergent zoonosis in several regions of the world and it is considered a public health problem. Trichinellosis represent one of the most important zoonotic diseases in Argentina. The purpose of this study was to develop a real-time PCR assay with internal control to detect Trichinella spiralis DNA in samples of swine muscles and its derivatives. PCR amplification of DNA from muscle samples was performed by real-time PCR with an internal porcine DNA amplification control. The developed PCR assay specifically detects T. spiralis showing no amplification with other Trichinella genotypes and showed an estimated sensitivity of 0.024 larvae per gram in swine muscle samples. From the 21 samples analyzed, five samples negative by enzymatic digestion were positive by real-time PCR, which demonstrates that this technique is capable of detecting T. spiralis in cases when enzymatic digestion cannot. In this work, a new molecular assay with internal control for T. spiralis detection was successfully developed.