Diagnosis of New World leishmaniasis: specific detection of species of the Leishmania braziliensis complex by amplification of kinetoplast DNA.
We have sequenced single kinetoplast DNA minicircles from three species and part of a minicircle from the fourth major species within the Leishmania braziliensis complex. Alignment of these sequences with each other and with those of other kinetoplastids allowed the selection of a pair of oligonucleotides suitable as primers in a polymerase chain reaction which is highly specific for the Leishmania braziliensis complex. The reaction is capable of detecting less than one femtogramme of kinetoplast DNA. It has been tested with crude specimens from South American leishmaniasis patients, potential wild animal reservoirs and sandfly vectors. The tests indicate that these primers are suitable for diagnosis of leishmaniasis and potentially useful in epidemiological surveys.
MRC Outstation of N.I.M.R., Molteno Laboratories, Cambridge, U.K.
M H
MH
de Bruijn
D C
DC
Barker
D000818:Animals; D001483:Base Sequence; D004270:DNA, Circular / Q000302:isolation & purification; D016054:DNA, Protozoan / Q000302:isolation & purification*; D005784:Gene Amplification; D006801:Humans; D007892:Leishmania braziliensis / Q000235:genetics / Q000302:isolation & purification*; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis* / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D004270:DNA, Circular; D016054:DNA, Protozoan