ABSTRACT
The aim of this work was to establish a modified pre-diagnostic polymerase chain reaction (PCR) protocol using a single primer set that enables successful amplification of a highly conserved mammalian sequence in order to determine overall sample DNA quality for multiple mammalian species that inhabit areas endemic for leishmaniasis. The gene encoding interphotoreceptor retinoid-binding protein (IRBP), but not other conserved genes, was efficiently amplified in DNA samples from tail skin, ear skin, bone marrow, liver and spleen from all of the species tested. In tissue samples that were PCR-positive for Leishmania, we found that DNA from 100 percent, 55 percent and 22 percent of the samples tested resulted in a positive PCR reaction for the IRBP, beta-actin and beta-globin genes, respectively. Nucleotide sequencing of an IRBP amplicon resolved any questions regarding the taxonomical classification of a rodent, which was previously based simply on the morphological features of the animal. Therefore, PCR amplification and analysis of the IRBP amplicon are suitable for pre-diagnostically assessing DNA quality and identifying mammalian species living in areas endemic to leishmaniasis and other diseases.
Subject(s)
Animals , Dogs , Actins , DNA, Protozoan , Eye Proteins , Leishmaniasis/veterinary , Polymerase Chain Reaction/veterinary , Retinol-Binding Proteins , beta-Globins , Actins , DNA Primers , Endemic Diseases , Eye Proteins , Leishmaniasis , Marsupialia , Polymerase Chain Reaction/methods , Rodentia , Retinol-Binding Proteins , beta-GlobinsSubject(s)
Humans , Animals , Antibodies, Protozoan/blood , Flow Cytometry/methods , Immunoglobulin G/blood , Leishmania braziliensis/immunology , Leishmaniasis, Cutaneous/diet therapy , Antibodies, Protozoan/immunology , Case-Control Studies , Fluorescent Antibody Technique, Indirect , Immunoglobulin G/immunology , Leishmaniasis, Cutaneous/immunology , Predictive Value of Tests , Sensitivity and SpecificityABSTRACT
Although cases of leishmaniasis co-infection have been described in acquired immunodeficiency syndrome patients as well as those who have undergone organ transplants, to our knowledge, the present report is the first documented case of simultaneous cutaneous, visceral and ocular leishmaniasis due to Leishmania (Viannia) braziliensis in a transplant patient. The patient had been using immunosuppressive drugs since receiving a transplanted kidney. The first clinical signs of leishmaniasis included fever, thoracic pain, hepatosplenomegaly, leucopenia and anemia. The cutaneous disease was revealed by the presence of amastigotes in the skin biopsy. After three months, the patient presented fever with conjunctive hyperemia, intense ocular pain and low visual acuity. Parasites isolated from iliac crest, aqueous humor and vitreous body were examined using a range of molecular techniques. The same strain of L. (V.) braziliensis was responsible for the different clinical manifestations. The immunosuppressive drugs probably contributed to the dissemination of Leishmania