Molecular identification of Leishmania spp. clinical isolates from Colombia based on hsp70 gene.
BACKGROUND
Leishmaniasis is highly prevalent in Colombia, where at least six different species can cause disease of varying clinical presentations in humans. The identification of the infecting species is quite important for prognosis, therapeutics and epidemiology. Different techniques with variable discriminatory power have been used for the identification.


OBJECTIVE
To carry out the molecular identification of Leishmania species through the amplification of a fragment of the hsp70 gene.


METHODS
Molecular amplification of the hsp70 gene fragment (PCR-hsp70) followed by restriction fragment length polymorphism analysis (RFLP) was done for identification purposes using DNA from 81 clinical isolates of Leishmania.


RESULTS
A single amplicon was obtained for all samples analyzed. The enzymatic restrictions of the 81 PCR products identified 70 with a banding pattern corresponding to L. braziliensis with two different patterns (62 and eight isolates, respectively), nine isolates compatible with L. panamensis and two with L. guyanensis. The geographical origin of the isolates is consistent with previous reports about the distribution of the corresponding species in Colombia.


CONCLUSIONS
The PCR-hsp70/RFLP technique used is a valid tool for the identification of Leishmania species isolated from clinical samples of patients in Colombia, which may also be applicable to the study of strains obtained from vectors and reservoirs with epidemiological significance.
Departamento de Parasitología, Instituto de Medicina Tropical Pedro Kourí, La Habana, Cuba. biomedica@ins.gov.co.
Ana M
AM
Montalvo
Jorge
J
Fraga
Ivón
I
Montano
Lianet
L
Monzote
Gert
G
Van der Auwera
Marcel
M
Marín
Carlos
C
Muskus
D000818:Animals; D003105:Colombia; D016054:DNA, Protozoan / Q000032:analysis* / Q000737:chemistry; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D007891:Leishmania / Q000737:chemistry*; D016773:Leishmaniasis, Cutaneous / Q000235:genetics / Q000469:parasitology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length
D016054:DNA, Protozoan; D018840:HSP70 Heat-Shock Proteins
Leishmania; diagnosis; polymerase chain reaction; polymorphism; restriction fragment length