Detection and identification of Leishmania species in field-captured phlebotomine sandflies based on mini-exon gene PCR

Paiva, Byanca Regina de; Secundino, Nágila Francinete Costa; Nascimento, João Cezar do; Pimenta, Paulo Filemon Paolucci; Galati, Eunice Aparecida Bianchi; Andrade, Hosana Aguiar Freitas de; Malafronte, Rosely S.

doi:10.1016/j.actatropica.2006.08.009

Cited by 75 publications

(60 citation statements)

References 33 publications

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“…To minimize possible errors and quantify the Leishmania infection/insect, a minimal infection rate (MR) was estimated using the formula MR=number of positive groups (pools) × 100 / number of total insects (PAIVA et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

Detection of Leishmania infantum in Lutzomyia longipalpis captured in Campo Grande, MS

Cunha

Andreotti

Cominetti

et al. 2014

Rev. Bras. Parasitol. Vet.

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Leishmaniasis is a zoonotic disease caused by protozoa of the genus Leishmania (Ross, 1903) and is the focus of considerable attention in human and veterinary medicine. In the city of Campo Grande, MS, the causative agent of visceral leishmaniasis is Leishmania infantum (= L. chagasi) primary vector, comprising approximately 92.9% of the local sandfly population, is Lutzomyia longipalpis. The aim of this work was to compare real-time PCR with PCR as a tool for the detection of the kinetoplast DNA (kDNA) of L. infantum in sandflies. Sandflies of this species were caught, and a total of 38 samples with 1-4 individuals in each sample were obtained; these were distributed across 13 districts and divided between seven urban areas of the city of Campo Grande, MS. Three positive samples were found by PCR and, when using real-time PCR, this was able to detect the presence of this agent in 6 of the 13 districts sampled, which were all located on the outskirts of the city, where indicates the greater enzootic potential of these regions, as they are closer to natural forest reserves. We conclude that real-time PCR can be used for epidemiological studies of L. infantum.Keywords: PCR, real-time PCR, phlebotomines, epidemiology. ResumoA Leishmaniose é uma zoonose causada por protozoários do gênero Leishmania (Ross 1903), objetos de considerável atenção em medicina humana e veterinária. Na cidade de Campo Grande -MS, o agente etiológico da Leishmaniose Visceral é Leishmania infantum (= L. chagasi), e o principal vetor é a espécie Lutzomyia longipalpis, que representa cerca de 92,9% da população de flebotomíneos. Este trabalho teve como objetivo avaliar a PCR em tempo real como ferramenta para a detecção de kDNA de L. infantum em flebotomíneos, comparando-se com PCR convencional. Flebotomíneos dessa espécie foram capturados, somando 38 amostras de 1 a 4 espécimens cada, distribuídas em 13 bairros, divididos entre as 7 regiões urbanas da cidade de Campo Grande -MS, e armazenados a -70 °C até a extração de ADN e amplificação por PCR e PCR em tempo real. Das 38 amostras testadas, foram encontradas 3 amostras positivas pela PCR convencional e 11 pela PCR em Tempo Real. Na otimização da PCR em tempo real, a temperatura de dissociação do amplificado foi de 82, 89 °C. Neste estudo, utilizando-se a técnica da PCR em tempo real, foi possível detectar a presença desse agente em 6 dos 13 bairros amostrados, todos na periferia da cidade, indicando o maior potencial enzoótico dessas regiões, que têm maior proximidade com reservas de matas naturais. Conclui-se que a PCR em tempo real pode ser utilizada para estudo epidemiológico de L. infantum.

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Section: Discussionmentioning

confidence: 99%

Detection of Leishmania infantum in Lutzomyia longipalpis captured in Campo Grande, MS

Cunha

Andreotti

Cominetti

et al. 2014

Rev. Bras. Parasitol. Vet.

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“…These studies are critical to better understand the dynamics of disease transmission and to identify new parasite-vector associations. 12 The detection of Leishmania in sand fly specimens has traditionally been based on the finding of flagellate forms in sand fly midgut dissections. However, this method requires advanced expertise 13 and is cumbersome and time-consuming.…”

Section: Introductionmentioning

confidence: 99%

Natural Leishmania Infection of Lutzomyia auraensis in Madre de Dios, Peru, Detected by a Fluorescence Resonance Energy Transfer–Based Real-Time Polymerase Chain Reaction

Valdivia

Santos²,

Fernández³

et al. 2012

The American Society of Tropical Medicine and Hygiene

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Abstract. Leishmania species of the Viannia subgenus are responsible for most cases of New World tegumentary leishmaniasis. However, little is known about the vectors involved in disease transmission in the Amazon regions of Peru. We used a novel real-time polymerase chain reaction (PCR) to assess Leishmania infections in phlebotomines collected in rural areas of Madre de Dios, Peru. A total of 1,299 non-blood fed female sand flies from 33 species were captured by using miniature CDC light traps. Lutzomyia auraensis was the most abundant species (63%) in this area. Seven of 164 pools were positive by PCR for Leishmania by kinetoplast DNA. The real-time PCR identified four Lu. auraensis pools as positive for L. (Viannia) lainsoni and L. (V.) braziliensis. The minimum infection prevalence for Lu. auraensis was estimated to be 0.6% (95% confidence interval = 0.20-1.42%). Further studies are needed to assess the importance of Lu. auraensis in the transmission of New World tegumentary leishmaniasis in hyperendemic areas of Peru.

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“…Several DNA targets such as the rRNA gene, minexon-derived RNA gene, repeated genome sequences and the kDNA minicircle (Aransay et al 2000, Paiva et al 2006) are used in the PCR detection and identification of Leishmania spp.…”

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confidence: 99%

The molecular detection of different Leishmania species within sand flies from a cutaneous and visceral leishmaniasis sympatric area in Southeastern Brazil

Saraiva

Filho

Silva

et al. 2010

Mem. Inst. Oswaldo Cruz

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Detection and identification of Leishmania species in field-captured phlebotomine sandflies based on mini-exon gene PCR

Cited by 75 publications

References 33 publications

Detection of Leishmania infantum in Lutzomyia longipalpis captured in Campo Grande, MS

Detection of Leishmania infantum in Lutzomyia longipalpis captured in Campo Grande, MS

Natural Leishmania Infection of Lutzomyia auraensis in Madre de Dios, Peru, Detected by a Fluorescence Resonance Energy Transfer–Based Real-Time Polymerase Chain Reaction

The molecular detection of different Leishmania species within sand flies from a cutaneous and visceral leishmaniasis sympatric area in Southeastern Brazil

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