Characterisation of nuclear ribosomal DNA sequences from Onchocerca volvulus and Mansonella ozzardi (Nematoda: Filarioidea) and development of a PCR-based method for their detection in skin biopsies.
The internal transcribed spacer region (ITS1, 5.8S gene and ITS2) of the two filarial nematodes Onchocerca volvulus and Mansonella ozzardi was sequenced, and two species-specific primers designed in the ITS2 to develop a PCR-based method for their specific detection and differentiation. When used with a universal reverse primer, the two species-specific primers gave amplification products of different size, which were readily separated in an agarose gel. The PCR was tested on skin biopsies from 51 people from three localities in Brazil where M. ozzardi is present, and results have been compared with those of parasitological examination of blood. The species-specific PCR gave a higher percentage of detection of infection by M. ozzardi than the parasitological examination of blood. No infection with O. volvulus was detected by PCR. This PCR-based assay may assist in determining the nature of infection in areas where both filarial species exist in sympatry.
Department of Entomology, The Natural History Museum, Cromwell Road, SW7 5BD, London, UK. r.morales-hojas@nhm.ac.uk
R
R
Morales-Hojas
R J
RJ
Post
A J
AJ
Shelley
M
M
Maia-Herzog
S
S
Coscarón
R A
RA
Cheke
D000818:Animals; D001706:Biopsy; D017931:DNA Primers; D018509:DNA, Helminth / Q000032:analysis / Q000097:blood; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D006801:Humans; D008366:Mansonella / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D008368:Mansonelliasis / Q000175:diagnosis / Q000469:parasitology; D008969:Molecular Sequence Data; D017181:Onchocerca volvulus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D009855:Onchocerciasis / Q000175:diagnosis / Q000469:parasitology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012340:RNA, Ribosomal, 5.8S / Q000235:genetics*; D017422:Sequence Analysis, DNA; D012867:Skin / Q000469:parasitology*; D013045:Species Specificity
D017931:DNA Primers; D018509:DNA, Helminth; D021903:DNA, Ribosomal Spacer; D012340:RNA, Ribosomal, 5.8S