Visceral leishmaniasis in Teresina, n. e. Brazil: towards a DNA probe kit and its adaptation to processing blood-contaminated samples.
The Lmet2 chemiluminescent DNA probe is a valuable tool for identifying parasites of the Leishmania donovani -complex in sand flies, dogs and human samples. Recent blood meals in sand flies or blood contamination of tissue samples inhibited probe sensitivity, whether radiolabelled or chemiluminescent detection systems were used. Treatment of membranes with protease before hybridisation restored positive signal. Alternatively samples could be lysed with protease and applied to membranes with a vacuum blotting apparatus. The Lmet2 protocol provides the basis for a DNA probe kit that is adaptable for use with a wide range of other probes.
McNerney
R
R
London School of Hygiene and Tropical Medicine, UK.
Frame
I A
IA
Vexenat
J A
JA
Fonseca de Castro
J A
JA
Howard
M K
MK
Dillon
R
R
Wilson
S
S
Miles
M A
MA
D000818:Animals; D015982:Bias; D001706:Biopsy; D002985:Clinical Protocols; D015342:DNA Probes; D016054:DNA, Protozoan; D004195:Disease Models, Animal; D004283:Dog Diseases / Q000473:pathology*; D004285:Dogs; D010450:Endopeptidases; D005069:Evaluation Studies as Topic; D007303:Insect Vectors / Q000469:parasitology*; D007893:Leishmania donovani; D018314:Leishmania infantum / Q000235:genetics*; D007898:Leishmaniasis, Visceral / Q000473:pathology*; D011576:Psychodidae / Q000469:parasitology*; D011933:Reagent Kits, Diagnostic / Q000592:standards*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods*
D015342:DNA Probes; D016054:DNA, Protozoan; D011933:Reagent Kits, Diagnostic; D010450:Endopeptidases