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Diagnosis of infections with Leishmania infantum using PCR–ELISA

Published online by Cambridge University Press:  12 July 2001

J. MARTIN-SANCHEZ ,
M. C. LOPEZ-LOPEZ ,
C. ACEDO-SANCHEZ ,
J. J. CASTRO-FAJARDO ,
J. A. PINEDA  and
F. MORILLAS-MARQUEZ
J. MARTIN-SANCHEZ
Affiliation:
Departamento de Parasitología, Facultad de Farmacia, C/Manuel Clavero s/n, Universidad de Granada, Campus Universitario de Cartuja, 18.071, Granada, Spain
M. C. LOPEZ-LOPEZ
Affiliation:
Instituto de Parasitología y Biomedicina ‘López Neyra', C/ Ventanilla 11, Consejo Superior de Investigaciones Científicas, 18.001, Granada, Spain
C. ACEDO-SANCHEZ
Affiliation:
Departamento de Parasitología, Facultad de Farmacia, C/Manuel Clavero s/n, Universidad de Granada, Campus Universitario de Cartuja, 18.071, Granada, Spain
J. J. CASTRO-FAJARDO
Affiliation:
Sociedad Protectora de Animales y Plantas ‘Francisco de Asís', Avda Badajoz s/n, Granada, Spain
J. A. PINEDA
Affiliation:
Grupo para el Estudio de Hepatitis Vírica y Sida, Servicio de Medicina Interna, Hospital Universitario Nuestra Señora de Valme, Carretera de Cádiz s/n, 41014 Sevilla, Spain
F. MORILLAS-MARQUEZ
Affiliation:
Departamento de Parasitología, Facultad de Farmacia, C/Manuel Clavero s/n, Universidad de Granada, Campus Universitario de Cartuja, 18.071, Granada, Spain
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Abstract

On the basis of partial amplification of a cloned fragment of kDNA of Leishmania infantum which is specific for this species, we developed a PCR–ELISA technique which avoids the problems associated with classical diagnostic techniques. This technique was tested on 33 L. infantum strains from 19 different zymodemes, which were recognized equally. It was also used on human and canine clinical samples. PCR–ELISA has a higher sensitivity than the other techniques used (IFAT, parasite cultures, optical microscopy of stained samples) and permits detection of a minimum of 0.1 promastigotes or 1 fg of genomic DNA. PCR–ELISA can be used to diagnose human cutaneous leishmaniasis using material obtained by scraping the lesion margin, and human visceral leishmaniasis in HIV(+) individuals and canine leishmaniasis with peripheral blood samples. The presence of L. infantum in dogs with low antibody titres with IFAT technique (20 and 40) was demonstrated indicating that seroprevalence data from epidemiological studies underestimate the true rates of infection.

Keywords

Leishmania infantumkinetoplast DNAPCR–ELISAhuman cutanean leishmaniasishuman visceral leishmaniasiscanine leishmaniasis
Type
Research Article
Information
Parasitology , Volume 122 , Issue 6 , June 2001 , pp. 607 - 615
Copyright
© 2001 Cambridge University Press

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