Differentiation of Trichinella genotypes by polymerase chain reaction using sequence-specific primers.
A method was developed to identify species and genotypes within the genus Trichinella using polymerase chain reaction (PCR) and specific primers. Enzymatic amplification of 2 partially conserved and repetitive genomic DNA sequences that have been shown to be variable in length within the different Trichinella genotypes form the basis of this test. Within these regions of the genome, 4 sets of primers were evaluated from which 2 were chosen for their ability to differentiate among the genotypes under stringent primer annealing conditions while maintaining high yields of amplification product. Differences in the size of PCR products from multiple isolates of each genotype indicate sufficient variation to identify 7 of the 8 parasite groups within this genus. One primer set can differentiate among some genotypes working from a single larva. Identification of Trichinella genotypes will assist in distinguishing between sylvatic and synanthropic life cycles. Such information will be critical in tracing sources of trichinellosis by easily and unambiguously identifying likely host reservoirs and will provide valuable information for instituting methods of control.
Appleyard
G D
GD
Centre for Animal Parasitology, Canadian Food Inspection Agency, Saskatoon.
Zarlenga
D
D
Pozio
E
E
Gajadhar
A A
AA
D000818:Animals; D017931:DNA Primers / Q000737:chemistry*; D018509:DNA, Helminth / Q000032:analysis*; D014644:Genetic Variation; D005838:Genotype; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D014234:Trichinella / Q000145:classification / Q000235:genetics*
D017931:DNA Primers; D018509:DNA, Helminth