Detection and identification of eight Trichinella genotypes by reverse line blot hybridization

• Published in: Journal of clinical microbiology Vol. 39; no. 2; pp. 642 - 646
• Main Authors: ROMBOUT, Y. B, BOSCH, S, VAN DER GIESSEN, J. W. B
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.02.2001
• Subjects: Animals; Biological and medical sciences; DNA, Helminth - genetics; DNA, Ribosomal - genetics; Fundamental and applied biological sciences. Psychology; Genotype; Host parasite relation; pathogenicity; Humans; Introns; Invertebrates; Nemathelminthia. Plathelmintha; Nucleic Acid Hybridization; Parasitology; RNA, Helminth - genetics; RNA, Ribosomal, 5S - genetics; Trichinella - classification; Trichinella - genetics; Trichinella - isolation & purification; Trichinella spiralis - genetics; Trichinella spiralis - isolation & purification; Trichinellosis - parasitology; Reverse line blot hybridization; Molecular hybridization; Genetic variability; Trichinella; Ribosomal DNA; Genotype; Spacer DNA; Identification; Species specificity; 5S-RNA; Polymerase chain reaction; Helmintha; Nemathelminthia; Invertebrata; Nematoda
• ISSN: 0095-1137; 1098-660X
• DOI: 10.1128/JCM.39.2.642-646.2001

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.