Trypanosoma vivax: characterization of the spliced-leader gene of a Brazilian stock and species-specific detection by PCR amplification of an intergenic spacer sequence.
The sequence of the spliced-leader gene repeat of a Brazilian Trypanosoma vivax stock from cattle showed high similarity to sequences of West African T. vivax in both intron and intergenic sequences. This is the first evidence based on DNA sequences of close-relatedness between Brazilian and West African T. vivax stocks. A T. vivax-specific diagnostic PCR assay based on spliced-leader gene intergenic sequences was able to amplify DNA from T. vivax stocks from South America (Brazil, Bolivia, and Colombia) and West Africa. Species-specificity of this method was confirmed by results obtained by testing 15 other trypanosomes, including other species and subspecies that can also infect cattle. The PCR assay developed presented high sensitivity, detecting the DNA content of only one parasite and also revealing T. vivax infection in asymptomatic animals without detectable parasitemia by microhematocrit or in Giemsa-stained blood smears. Use of crude preparations from field-blood samples collected on both filter paper and glass slides as DNA template suggested that this method could be useful for the diagnosis of T. vivax in large epidemiological studies.
Ventura
R M
RM
Departamento de Parasitologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, 05508-900, Brazil.
Paiva
F
F
Silva
R A
RA
Takeda
G F
GF
Buck
G A
GA
Teixeira
M M
MM
D000818:Animals; D001483:Base Sequence; D001938:Brazil; D002417:Cattle; D015342:DNA Probes / Q000592:standards; D021901:DNA, Intergenic / Q000737:chemistry / Q000235:genetics*; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D020038:RNA, Spliced Leader / Q000235:genetics*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D012756:Sheep; D013045:Species Specificity; D016689:Trypanosoma vivax / Q000145:classification / Q000235:genetics*
D015342:DNA Probes; D021901:DNA, Intergenic; D016054:DNA, Protozoan; D020038:RNA, Spliced Leader