[Show abstract][Hide abstract] ABSTRACT: To identify the aetiological agents of cutaneous leishmaniasis and to investigate the genetic polymorphism of Leishmania (Viannia) parasites circulating in an area with endemic cutaneous leishmaniasis (CL) in the Atlantic rainforest region of northeastern Brazil.
Leishmania spp. isolates came from three sources: (i) patients diagnosed clinically and parasitologically with CL based on primary lesions, secondary lesions, clinical recidiva, mucocutaneous leishmaniasis and scars; (ii) sentinel hamsters, sylvatic or synanthropic small rodents; and (iii) the sand fly species Lutzomyia whitmani. Isolates were characterised using monoclonal antibodies, multilocus enzyme electrophoresis (MLEE) and polymerase chain reaction-restriction fragment length polymorphism of the internal transcribed spacer region rDNA locus.
Seventy-seven isolates were obtained and characterised. All isolates were identified as Leishmania (Viannia) braziliensis serodeme 1 based on reactivity to monoclonal antibodies. MLEE identified 10 zymodemes circulating in the study region. Most isolates were classified as zymodemes closely related to L. (V.) braziliensis, but five isolates were classified as Leishmania (Viannia) shawi. All but three of the identified zymodemes have so far been observed only in the study region. Enzootic transmission and multiclonal infection were observed.
Our results confirm that transmission cycle complexity and the co-existence of two or more species in the same area can affect the level of genetic polymorphism in a natural Leishmania population. Although it is not possible to make inferences as to the modes of genetic exchange, one can speculate that some of the zymodemes specific to the region are hybrids of L. (V.) braziliensis and L. (V.) shawi.
Tropical Medicine & International Health 09/2009; 14(10):1278-86. DOI:10.1111/j.1365-3156.2009.02361.x · 2.33 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: It is uncertain whether Leishmania parasites ever disappear after clinical cure of American cutaneous leishmaniasis (ACL). Recently, sensitive molecular techniques have allowed the identification of Leishmania parasites directly in specimens from patients' scars.
Scars of 32 patients from northeastern Brazil who were treated and clinically cured of ACL were analyzed by use of polymerase chain reaction (PCR), culture, and histopathologic examination.
DNA specific for Leishmania (Viannia) was detected in scars of 30 (93.7%) of 32 patients. In specimens from 3 of the scars, Leishmania parasites could be isolated by culture; PCR results also were positive for those 3 specimens. No parasites were found by histopathologic examination, and fibrotic alterations were present in all cases, with slight inflammatory foci observed in 4 of the cases studied.
The results suggest that clinical cure of ACL is rarely associated with sterile cure. The implications of persistence of parasites for the clinical evolution, relapse, and transmission of leishmaniasis deserves further studies, particularly with the increasing incidence of coinfection with leishmaniasis and acquired immunodeficiency syndrome.
The Journal of Infectious Diseases 04/2004; 189(6):1018-23. DOI:10.1086/382135 · 6.00 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: PCR-based approaches targeting kinetoplast DNA were evaluated for the diagnosis of American cutaneous leishmaniasis (ACL)
in regions of endemicity in northeastern Brazil. A total of 119 cutaneous biopsy specimens from patients with ACL and nonleishmaniasis
cutaneous lesions were studied. Two PCR-based systems were used; one was specific for the subgenus Viannia, and the other was specific for the genus Leishmania. The PCR specific for the subgenus Viannia had a sensitivity of 95.4%, whereas the genus-specific PCR detected the target DNA in 88.2% of the samples tested. The specificities
of the assays, determined with samples from a group with nonleishmaniasis cutaneous lesions, was 100%. The results of the
conventional tests indicate that the sensitivities of the PCR-based methods were significantly higher than those of smear
examination, histological staining, and isolation by culture (P < 0.05). Antibodies specific for Leishmania braziliensis were detected by indirect immunofluorescence in 82.9% of the patients tested. Parasites were isolated from 40 of 86 patients
(46.5%). Sixty-seven percent of dermal scrapings and 66.2% of stained tissue sections were positive by microscopy. Amplified
products from the subgenus-specific PCR hybridized with the Leishmania panamensis minicircle, confirming infection consistent with L. braziliensis. The evidence available at present incriminates L. braziliensis as the only causative agent of ACL in the state of Pernambuco in Brazil.
[Show abstract][Hide abstract] ABSTRACT: The antigen specificity and the level of the antibody response were analysed in Perambuco State, Brazil, in sera collected in 1995-96 from 58 patients with clinical American cutaneous leishmaniasis (ACL), 25 ACL patients with apparent cure after chemotherapy with meglumine antimonate, and 10 ACL patients with spontaneous cure. Assessment was by immunoblot analysis, ELISA and indirect immunofluorescence, with Leishmania (Viannia) braziliensis antigens, with a particular interest in evaluating whether the dynamics of the antibody response could be useful to monitor clinical cure. A clear decrease of IgG antibody reactivity was noticed after clinical healing, for all of the antigens analysed, with the exception of the 19 kDa antigen, whose recognition frequency in fact increased in the spontaneously cured patients, suggesting that this antigen may play a role in protective immunity against cutaneous leishmaniasis. The recognition frequencies of the most frequently recognized antigens (27 and 30 kDa antigens) diminished approximately 2-fold in patients clinically healed, suggesting that they could be useful as a marker of cure of ACL. In addition, some of the healthy individuals living in endemic areas presented the same immunoblotting pattern of reactivity observed in active ACL, possibly representing asymptomatically infected individuals.
Transactions of the Royal Society of Tropical Medicine and Hygiene 03/2001; 95(2):203-6. DOI:10.1016/S0035-9203(01)90168-3 · 1.84 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The antibody response in patients with American cutaneous leishmaniasis was analyzed by immunoblotting with soluble and insoluble
antigens of Leishmania braziliensis. The recognition of the 27- and/or 30-kDa soluble antigens was considered relevant for the diagnosis of cutaneous leishmaniasis.
Immunoblotting was found to be significantly more sensitive and specific than indirect immunofluorescence and enzyme-linked