A PCR based assay for detection and differentiation of African trypanosome species in blood.
Direct PCR analysis of trypanosome infected blood samples in the quantities required for large scale epidemiological study has always been problematic. Current methods for identifying and differentiating trypanosomes typically require several species-specific reactions, many of which rely on mouse passaged samples to obtain quality concentrated genomic DNA. As a consequence important epidemiological information may be lost during the sample preparation stage. Here, we report a PCR methodology that reduces processing and improves on the sensitivity of present screening methods. The PCR technique targets the gene encoding the small ribosomal subunit in order to identify and differentiate all clinically important African trypanosome species and some subspecies. The method is more economical, simple, and sensitive than current screening methods, and yields more detailed information, thereby making it a viable tool for large-scale epidemiological studies.
Cox
Andrew
A
Centre for Tropical Veterinary Medicine, Royal (Dick) School of Veterinary Medicine, University of Edinburgh, Easter Bush, Roslin, Midlothian EH25 9RG, Scotland, UK.
Tilley
Aimee
A
McOdimba
Francis
F
Fyfe
Jenna
J
Eisler
Mark
M
Hide
Geoff
G
Welburn
Susan
S
D000818:Animals; D002417:Cattle; D021903:DNA, Ribosomal Spacer / Q000737:chemistry; D006801:Humans; D018512:Parasitemia / Q000469:parasitology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012335:RNA, Ribosomal / Q000235:genetics; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D013045:Species Specificity; D014345:Trypanosoma / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D014353:Trypanosomiasis, African / Q000469:parasitology*; D014354:Trypanosomiasis, Bovine / Q000469:parasitology*
D021903:DNA, Ribosomal Spacer; D012335:RNA, Ribosomal