Design of primer pairs for species-specific diagnosis of Leishmania (Leishmania) infantum chagasi using PCR

Neto, Osvaldo José da Silveira; Duarte, Sabrina Castilho; Costa, Hérika Xavier da; Linhares, Guido Fontgalland Coelho

doi:10.1590/s1984-29612012000300024

Cited by 5 publications

(3 citation statements)

References 18 publications

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“…The sensitivity obtained (0.2 fg) was similar to the related by OLIVEIRA et al 25 (2 fg) and DE BRUJIN & BARKER 11 (1 fg) but it was higher than the one related by SILVEIRA NETO et al 31 (1 pg/μL) in a PCR for L. infantum . The high sensitivity was checked by the positive results obtained with low concentrations of DNA from L. infantum , demonstrating the ability to detect parasite DNA in peripheral blood and lesions, even in situations where the number of parasites is low.…”

Section: Discussionsupporting

confidence: 86%

NEW PRIMERS FOR DETECTION OF Leishmania infantum USING POLYMERASE CHAIN REACTION

Gualda

Marcussi

Neitzke-Abreu

et al. 2015

Rev. Inst. Med. trop. S. Paulo

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SUMMARY Leishmania infantum causes visceral leishmaniasis (VL) in the New World. The diagnosis of VL is confirmed by parasitological and serological tests, which are not always sensitive or specific. Our aim was to design new primers to perform a Polymerase Chain Reaction (PCR) for detecting L. infantum. Sequences of the minicircle kinetoplast DNA (kDNA) were obtained from GenBank, and the FLC2/RLC2 primers were designed. Samples of DNA from L. infantum, Leishmania amazonensis,Leishmania braziliensis, Leishmania guyanensis, Leishmania naiffi, Leishmania lainsoni, Leishmania panamensis,Leishmania major and Trypanosoma cruzi were used to standardize the PCR. PCR with FLC2/RLC2 primers amplified a fragment of 230 bp and the detection limit was 0.2 fg of L. infantum DNA. Of the parasite species assayed, only L. infantum DNA was amplified. After sequencing, the fragment was aligned to GenBank sequences, and showed (99%) homology with L. infantum. In the analysis of blood samples and lesion biopsy from a dog clinically suspected to have VL, the PCR detected DNA from L. infantum. In biopsy lesions from humans and dogs with cutaneous leishmaniasis, the PCR was negative. The PCR with FLC2/RLC2 primers showed high sensitivity and specificity, and constitutes a promising technique for the diagnosis of VL.

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Section: Discussionsupporting

confidence: 86%

NEW PRIMERS FOR DETECTION OF Leishmania infantum USING POLYMERASE CHAIN REACTION

Gualda

Marcussi

Neitzke-Abreu

et al. 2015

Rev. Inst. Med. trop. S. Paulo

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“…From a public health perspective and considering the control of visceral leishmaniasis, this would be relevant in order to avoid the unnecessary culling of L. braziliensis-infected dogs (MADEIRA et al, 2006). In this regard, the use of specific PCR-based methods should be advisable to distinguish dogs infected by L. braziliensis from those infected by L. infantum (SILVEIRA NETO et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Clinical and hematological findings in Leishmania braziliensis-infected dogs from Pernambuco, Brazil

Figueredo

Paiva-Cavalcanti

Almeida

et al. 2012

Rev. Bras. Parasitol. Vet.

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Canine cutaneous leishmaniasis by Leishmania braziliensis is a neglected, but widespread disease of dogs in South America. This paper describes clinical and hematological alterations in 17 L. braziliensis-infected dogs from Brazil. The most common hematological findings were thrombocytopenia (82.4%), anemia (70.6%), low packed cell volume (52.9%) and eosinophilia (41.2%). Twelve (70.6%) dogs displayed at least one evident physical alteration; 11 dogs (64.7%) presented skin lesions, four (23.5%) had weight loss and two (11.8%) onychogryphosis. L. braziliensis-infected dogs present clinical and hematological signs often observed in dogs infected by other pathogens. This indicates that veterinarians and public health workers should not consider the presence of non-specific clinical signs as diagnostic criteria for visceral leishmaniasis in dogs living endemic areas to avoid misdiagnosis and subsequent elimination of dogs infected by L. braziliensis.Keywords: Leishmania braziliensis, dogs, Pernambuco, Brazil. ResumoA leishmaniose cutânea canina causada por Leishmania braziliensis é uma doença negligenciada, mas disseminada entre cães na America do Sul. Este artigo descreve alterações clínicas e hematológicas em 17 cães infectados por L. braziliensis do Brasil. As alterações hematológicas mais comuns foram trombocitopenia (82,4%), anemia (70,6%), baixo valor de hematócrito (52,9%) e eosinofilia (41,2%). Doze (70,6%) cães apresentaram pelo menos uma alteração física; 11 (64,7%) apresentaram lesões cutâneas, quatro (23,5%) perda de peso e dois (11,8%) onicogrifose. Cães infectados por L. braziliensis apresentaram alterações clínicas e hematológicas inespecíficas que são comumente observadas em cães infectados por outros patógenos. Isso indica que veterinários e profissionais de saúde pública não deveriam considerar a presença de tais sinais clínicos como critério de diagnóstico para leishmaniose visceral em cães, em áreas endêmicas, no intuito de evitar um diagnóstico equivocado e a subsequente eliminação de cães infectados por L. braziliensis.

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“…For Leishmania infantum DNA, species‐specific primers were used for the 18S ribosomal RNA (rRNA) region, LCS1 (5′‐GCAATGCCAGCTACATATATG‐3′) and LCS3 (5′‐CAGCTTTTTGGGTGGGTAACA‐3′) (Neto et al, 2012). For amplification of Leishmania braziliensis kDNA, species‐specific primers B1 (5′‐GGGGTTGGTGTAATATAGTGG‐3′) and B2 (5′‐CTAATTGTGCACGGGGAGG‐3′) as developed by De Bruijn and Barker (1992), using the PCR methodology of Alexander et al (1998).…”

Section: Methodsmentioning

confidence: 99%

Identification of Leishmania infantum and Leishmania braziliensis in captive primates from a zoo in Brazil

Guiraldi

Santos

Manzini

et al. 2022

American J Primatol

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Wild nonhuman primates (NHP) are considered natural hosts of a protozoan parasite from the genus Leishmania, the etiological agent of leishmaniasis. It is important to study the population of this infectious agent in zoo animals to establish surveillance and control mechanisms in Sorocaba through the application of a One Health approach, this is where human–animal–environment health and disease interface and can aid in the protection of endangered species. This study aimed to identify Leishmania infantum and Leishmania braziliensis in NHP living in a city where leishmaniasis is endemic. DNA was extracted from 48 NHP and analyzed using polymerase chain reaction primers that are specific for the species L. infantum and L. braziliensis. The results of our research revealed the first report of L. infantum and L. braziliensis naturally infecting primates at Sorocaba zoo. One primate from the species Plecturocebus vieirai was positive for L. infantum and five primates (four Alouatta caraya and one Ateles chamek) were positive for L. braziliensis. This indicates a possible role of these animals on the maintenance of these parasites.

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Design of primer pairs for species-specific diagnosis of Leishmania (Leishmania) infantum chagasi using PCR

Cited by 5 publications

References 18 publications

NEW PRIMERS FOR DETECTION OF Leishmania infantum USING POLYMERASE CHAIN REACTION

NEW PRIMERS FOR DETECTION OF Leishmania infantum USING POLYMERASE CHAIN REACTION

Clinical and hematological findings in Leishmania braziliensis-infected dogs from Pernambuco, Brazil

Identification of Leishmania infantum and Leishmania braziliensis in captive primates from a zoo in Brazil

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