Specific uni- and triplex-PCR for detection of single and three tapeworms transmitted from food to humans including beef and pork meat (cysticerci) (food-borne zoonoses) are needed to be developed. The authors have developed and tested diagnostic kit(s) for single and multiple, i.e. pig (Taenia solium); cattle (T. saginata) and human (T. asiatica) tapeworms, using mitochondrial genetic markers. The reverse primer (namely, TASSR) common for three species, which was used to pair with the forward primer specific to each species in PCR reaction. The PCR product for T. solium (with forward primer, TSOF) is 474 bp, for T. saginata (with TSAF) is 609 bp; and for T. asiatica (with T ASF) is 706 bp. Uniplex-PCR examined with single DNA template and species specific primers, and with crossing templates and crossing primers, generated highly specific products. Based on the length difference of the PCR products specific to each tapeworm, a triplex-PCR kit (designated as D(Tso+Tsa+Tas)) was developed and tested with a mix of three genomic DNA templates and with a mix of primers for two or three species, repectively. As results, triplex-PCR kit, namely kit D(Tso+Tsa+Tas) had very high specificity and sensitivity. It revealed that only the target species- specific primers could generate PCR product from the target species in the template-mix. Sensitivity of the kit allows PCR to obtain product with the limit of diluted genomic DNA template mix of three species as little as 0.8 ng. This kit is accurate, easy performing and sensitive, which can detect any target Taenia species among the above three species in a form of cysticerci in food (meat) and adult worms in humans and animals.
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