Detection of DNA from Leishmania (Viannia): accuracy of polymerase chain reaction for the diagnosis of cutaneous leishmaniasis.

Cutaneous leishmaniasis (CL) can occur in skin and mucosa, causing disfiguring lesions. The laboratory diagnosis of CL involves immunological methods and optical detection of the parasite, al of which have limitations. There is a need for more effective diagnostic methods for CL which wil allow treatment to be initiated more promptly in order to help prevent the development of severe forms of mucosal disease, and to estimate the prognosis of the infection. The polymerase chain reaction (PCR) has been widely used to diagnose CL, because of its higher sensitivity. This study estimated the accuracy and compared PCRs of samples from lesion scarification (PCR-L) and blood sample-enriched leukocytes (PCR-B) with three conventional diagnostic techniques: parasite direct search (DS), Montenegro skin test (MST), and indirect immunofluorescence reaction (IIF). The study included 276 patients under suspicion of CL. We conducted a cross-sectional study, in which patients were selected by convenience sampling. We used MP3H/MP1L primers to generate a Leishmania (Viannia) (minicircle kDNA) fragment of 70-bp. Of 106 patients with CL, 83.87%, 51.67%, 64.52%, 85.71%, or 96.10% tested positive by PCR-L, PCR-B, DS, IIF, or MST, respectively. Five patients tested positive only by PCR-L, and two other patients only by PCR-B. PCR-L is indicated for use in patients with chronic lesions or Leishmania reinfection, which may progress to mucosal lesion. PCR-B is indicated for use in patients with negative results in conventional tests or for patients with no apparent lesion. PCR is not only useful in diagnosing CL but also helps to identify the infecting species.

Environmental and animal characteristics as factors associated with American cutaneous leishmaniasis in rural locations with presence of dogs, Brazil.

The aim of the study was to investigate the importance of dogs, other domesticated animals and environmental characteristics as risk factors in the epidemiology of American cutaneous leishmaniasis (ACL). A retrospective survey of cases of human ACL in the last ten years and visits to homes in rural locations were carried out in the municipality of Arapongas (southern Brazil) from 2008 to 2010. ACL in humans was significantly associated with a distance of up to 25 meters from the residence to a forest area (OR 5.08; 95% CI: 1.35-21.04), undergrowth area (OR 6.80; 95% CI: 1.69-45.33) and stream (OR 5.87; 95% CI: 1.15-24.59); banana plants near the residence (OR 5.98; 95% CI: 1.49-39.84), absence of ceiling below the roof in the residence (OR 7.30; 95% CI: 1.26-158.1), the dumping of trash in the forest area (OR 26.33; 95% CI: 7.32-93.46) and presence of ACL in dogs in the surrounding area (OR 4.39; 95% CI: 1.37-13.45). In dogs, ACL was associated with a distance of 25 to 50 meters and 51 to 100 meters, respectively, from the residence to a forest area (OR 2.59; 95% CI: 1.08-5.98; OR 3.29; 95% CI: 1.64-6.62), the presence of a stream up to 25 m from the residence (OR 6.23; 95% CI: 2.34-16.54) and banana plants near the residence (OR 0.45; 95% CI: 0.25-0.80). In the locations studied in the municipality of Arapongas (Brazil), the results reveal that canine infection increases the risk of human infection by ACL and the characteristics surrounding the residence increase the risk of infection in both humans and dogs. Thus, integrated environmental management could be a useful measure to avoid contact between humans and phlebotomines.

Natural infection of Nyssomyia neivai by Leishmania (Viannia) spp. in the state of Paraná, southern Brazil, detected by multiplex polymerase chain reaction.

Natural sandfly infection by Leishmania spp. in an area endemic for American cutaneous leishmaniasis was analyzed using multiplex polymerase chain reaction (PCR). The sandflies were captured using Falcão light traps in an endemic area of the municipality of Doutor Camargo during March, April, and June 2008. In total, 1803 females were analyzed; 1755 were Nyssomyia neivai (Pinto) and 48 were Nyssomyia whitmani (Antunes and Coutinho). Multiplex PCR analyses using MP3H-MP1L and 5Llcac-3Llcac primers showed the presence of Leishmania (Viannia) spp. in 4/181 pools of sandflies, all Ny. neivai, that is, a minimal infection rate of 0.22%. This study showed, for the first time, the presence of DNA of Leishmania (Viannia) spp. in Ny. neivai. This suggests the existence of natural infection by Leishmania (Viannia) spp. in Ny. neivai in the state of Paraná. Multiplex PCR is an important tool in the detection of Leishmania infection in sandflies.

[American cutaneous leishmaniasis in municipalities in the northwestern region of Paraná State: use of remote sensing for analysis of vegetation types and places with disease occurrence]. / Leishmaniose tegumentar americana em municípios da região noroeste do estado do Paraná: utilização de sensoriamento remoto para análise do tipo de vegetação e os locais de ocorrência da doença.

American cutaneous leishmaniasis, an endemic disease in the northwestern region of Paraná, Brazil, is transmitted by phlebotomines to man and animals like dogs, armadillos, opossums and wild rodents. This disease has been occurring in places where forests have been felled and on the banks of rivers and lakes with arboreal vegetation, where man comes into contact with infected insects. This disease is a public health problem because of the occurrence of epidemic outbreaks throughout Paraná. Because of the importance of finding out about endemic areas, this study used epidemiological file data on patients attended between 1999 and 2004. These data were correlated with areas of forest felling that were identified by means of satellite remote sensing techniques and products. The results showed that the occurrences of cases in the municipalities of this region coincided with the presumed likely areas for patient infection.

Leishmaniose tegumentar americana em municípios da região noroeste do estado do Paraná: utilização de sensoriamento remoto para análise do tipo de vegetação e os locais de ocorrência da doença / American cutaneous leishmaniasis in municipalities in the northwestern region of Paraná State: use of remote sensing for analysis of vegetation types and places with disease occurrence

A leishmaniose tegumentar americana, doença endêmica da região noroeste do Paraná, é transmitida por flebotomíneos ao homem e a animais como cães, tatu, gambá e roedores silvestres. A doença vem ocorrendo em locais de derrubadas de matas e às margens de rios e lagoas com vegetação arbórea, onde o homem entra em contato com o inseto infectado. Esta doença constitui-se num problema de saúde pública devido à ocorrência de surtos epidêmicos em todo o Paraná. Em decorrência da importância de se conhecer áreas endêmicas, este trabalho utilizou dados de fichas epidemiológicas de pacientes atendidos no período de 1999 a 2004 associado às áreas de desmatamento identificadas por técnicas e produtos de sensoriamento remoto por satélite. Os resultados mostraram que a ocorrência de casos em municípios da região coincidem com as prováveis áreas supostas de infecção dos pacientes.

American cutaneous leishmaniasis, an endemic disease in the northwestern region of Paraná, Brazil, is transmitted by phlebotomines to man and animals like dogs, armadillos, opossums and wild rodents. This disease has been occurring in places where forests have been felled and on the banks of rivers and lakes with arboreal vegetation, where man comes into contact with infected insects. This disease is a public health problem because of the occurrence of epidemic outbreaks throughout Paraná. Because of the importance of finding out about endemic areas, this study used epidemiological file data on patients attended between 1999 and 2004. These data were correlated with areas of forest felling that were identified by means of satellite remote sensing techniques and products. The results showed that the occurrences of cases in the municipalities of this region coincided with the presumed likely areas for patient infection.