A single, multiplex PCR for differentiating all species of Trichinella

The genus Trichinella is currently divided into seven species and at least three additional, unclassified genotypes, Trichinella T6, T8 and T9, where both T8 and T9 have been deemed very similar to T. britovi. Other than for the non-encapsulated species, the absence of distinguishing morphological c haracters and the overlapping nature of the biological characters within th is genus make these traits unsuitable for diagnosis. Consequently, we have developed a simple PCR test for the unequivocal differentiation of all curr ently recognized species of Trichinella including Trichinella T6. DNA seque nce data from each Trichinella genotype were generated from infernal transc ribed spacers, ITS1 and ITS2, and from expansion segment V (ESV) of the rDN A repeat, from which five different PCR primer sets were chosen. When used simultaneously, this primer mix generates a simple and unique electrophoret ic DNA banding pattern for each species and genotype. The ESV-derived prime r set contributes at least one band to each agarose gel-derived genotypic p attern and therefore functions as an internal control for PCR integrity. Ge ographical isolates of each Trichinella genotype were used to verify the re liability and reproducibility of respective DNA banding patterns using sing le muscle larvae.