ABSTRACT
The genetic diversity within the genus Trichinella was studied using cleavage fragment length polymorphism (CFLP) analysis. The CFLP method generates specific fingerprints based on single nucleotide mutations. By this method the amplified intergenic regions of the 5S rRNA genes of the eight different genotypes of Trichinella were analysed. The CFLP pattern of T. spiralis was completely different compared with the sylvatic species T. britovi, T. nativa, T. nelsoni, and the genotypes Trichinella T5, Trichinella T6 and Trichinella T8. The T. pseudospiralis intergenic region can be differentiated by size from the other species of Trichinella.
Subject(s)
Genetic Variation , Polymorphism, Genetic , Trichinella/genetics , Animals , DNA Fingerprinting , DNA, Helminth/genetics , DNA, Helminth/isolation & purification , DNA, Ribosomal/genetics , Polymerase Chain Reaction , Reproducibility of ResultsABSTRACT
A reverse line blot (RLB) assay was developed to identify different Trichinella genotypes. The RLB assay accomplishes detection and specific identification of the different Trichinella genotypes and relies on hybridization of the amplified 5S ribosomal DNA intergenic spacer regions to specific, membrane-bound oligonucleotide probes. After one single amplification, we were able to detect and genetically identify six sibling species, i.e., T. spiralis, T. britovi, T. nativa, T. murrelli, T. nelsoni, and T. pseudospiralis, and two additional Trichinella genotypes, T6 and T8. Twenty-four Trichinella strains of different genotypes were unequivocally identified evaluated using one simple PCR-based assay based on single larvae. This assay allows the specific identification of Trichinella species without the need to passage larvae in laboratory animals.
Subject(s)
Trichinella/genetics , Trichinellosis/parasitology , Animals , DNA, Helminth/genetics , DNA, Ribosomal/genetics , Genotype , Humans , Introns , Nucleic Acid Hybridization , RNA, Helminth/genetics , RNA, Ribosomal, 5S/genetics , Trichinella/classification , Trichinella/isolation & purification , Trichinella spiralis/genetics , Trichinella spiralis/isolation & purificationABSTRACT
Echinococcus multilocularis was demonstrated in 5 out of 272 foxes in The Netherlands close to the border with Germany and Belgium. Besides microscopic examination of mucosal scrapings, two different PCR assays were used based on the detection of E. multilocularis DNA in colon content. Two distinct areas in The Netherlands were positive for E. multilocularis. Two positive foxes were found in the northern province of Groningen and three positive foxes were found in the southern province of Limburg. Both PCR assays detected more positive foxes compared to microscopic examination of the intestinal content. This is the first report of E. multilocularis in foxes occurring in The Netherlands.
