Accordance and concordance of PCR and NASBA followed by oligochromatography for the molecular diagnosis of Trypanosoma brucei and Leishmania.
OBJECTIVE
To evaluate the repeatability and reproducibility of four simplified molecular assays for the diagnosis of Trypanosoma brucei spp. or Leishmania ssp. in a multicentre ring trial with seven participating laboratories.


METHODS
The tests are based on PCR or NASBA amplification of the parasites nucleic acids followed by rapid read-out by oligochromatographic dipstick (PCR-OC and NASBA-OC).


RESULTS
On purified nucleic acid specimens, the repeatability and reproducibility of the tests were Tryp-PRC-OC, 91.7% and 95.5%; Tryp-NASBA-OC, 95.8% and 100%; Leish-PCR-OC, 95.9% and 98.1%; Leish-NASBA-OC, 92.3% and 98.2%. On blood specimens spiked with parasites, the repeatability and reproducibility of the tests were Tryp-PRC-OC, 78.4% and 86.6%; Tryp-NASBA-OC, 81.5% and 89.0%; Leish-PCR-OC, 87.1% and 91.7%; Leish-NASBA-OC, 74.8% and 86.2%.


CONCLUSIONS
As repeatability and reproducibility of the tests were satisfactory, further phase II and III evaluations in clinical and population specimens from disease endemic countries are justified.
Mugasa
Claire M
CM
Department of Veterinary Parasitology and Microbiology, Makerere University, Kampala, Uganda.
Deborggraeve
Stijn
S
Schoone
Gerard J
GJ
Laurent
Thierry
T
Leeflang
Mariska M
MM
Ekangu
Rosine A
RA
El Safi
Sayda
S
Saad
Al-farazdag A
AF
Basiye
Frank L
FL
De Doncker
Simone
S
Lubega
George W
GW
Kager
Piet A
PA
Büscher
Philippe
P
Schallig
Henk D F H
HD
D000818:Animals; D016054:DNA, Protozoan / Q000032:analysis; D006801:Humans; D007893:Leishmania donovani / Q000235:genetics / Q000302:isolation & purification*; D007898:Leishmaniasis, Visceral / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D021161:Self-Sustained Sequence Replication / Q000379:methods*; D013048:Specimen Handling / Q000379:methods; D014347:Trypanosoma brucei gambiense / Q000235:genetics / Q000302:isolation & purification*; D014353:Trypanosomiasis, African / Q000175:diagnosis*
D016054:DNA, Protozoan