Detection of Leishmania (L.) infantum in stray dogs by molecular techniques with sensitive species-specific primers

Rodrigo C. Silva; Virgínia B. Richini-Pereira; Mariana Kikuti; Pâmela M. Marson; Helio Langoni

doi:10.1080/01652176.2016.1252073

Detection of Leishmania (L.) infantum in stray dogs by molecular techniques with sensitive species-specific primers

Rodrigo C. Silva, Virgínia B. Richini-Pereira, Mariana Kikuti, Pâmela M. Marson, Helio Langoni

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

Abstract

Background: Canine visceral leishmaniasis (CVL) is a worldwide parasitic zoonosis caused by Leishmania (Leishmania) infantum around the world. Canids are the definitive hosts and sand flies the intermediate hosts. Objective: To test the hypothesis that a new species-specific primers (Lch14:Lch15, targeting a multiple alignment for L. infantum kDNA minicircle) is an efficient diagnostic tool for L. infantum. Methods: The presence of L. infantum DNA was assessed in blood samples of 69 stray dogs using the conventional PCR (cPCR) and quantitative PCR (qPCR). Additional 50 lymph nodes and 50 bone marrow samples (positive and negative samples for parasitological tests) from dogs from endemic and nonendemic areas for CVL were also used. Results: L. infantum strains, and all positive lymph node and bone marrow samples for parasitological test gave positive results for cPCR and qPCR, presenting analytical sensitivity of ∼10° parasite mL⁻¹. For the blood samples, 40/69 (58%; CI 95%; 46%-69%) resulted positive for L. infantum in both tests. All positive samples were confirmed by sequencing. Conclusion: This study showed the importance of the specific detection of L. infantum based on species-specific primers by molecular techniques, highlighting the application as a confirmation method in epidemiological studies and to adopt the best control measures.

Original language	English (US)
Pages (from-to)	23-30
Number of pages	8
Journal	Veterinary Quarterly
Volume	37
Issue number	1
DOIs	https://doi.org/10.1080/01652176.2016.1252073
State	Published - Nov 1 2017
Externally published	Yes

Bibliographical note

Funding Information:
We thank Oswaldo Cruz Foundation, Leishmania Collection of the Oswaldo Cruz Institute (CLIOC), and Trypanosoma Collection of Wild and Domestic Mammals, and Vectors (ColTryp) for providing all 10 panels of Leishmania spp., and Trypanosoma cruzi strains, and S?o Paulo State University (UNESP) for the logistical support.

Publisher Copyright:
© 2016 Informa UK Limited, trading as Taylor & Francis Group.

Keywords

Canine
KDNA
Leishmania infantum
PCR
Stray dogs

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10.1080/01652176.2016.1252073

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title = "Detection of Leishmania (L.) infantum in stray dogs by molecular techniques with sensitive species-specific primers",

abstract = "Background: Canine visceral leishmaniasis (CVL) is a worldwide parasitic zoonosis caused by Leishmania (Leishmania) infantum around the world. Canids are the definitive hosts and sand flies the intermediate hosts. Objective: To test the hypothesis that a new species-specific primers (Lch14:Lch15, targeting a multiple alignment for L. infantum kDNA minicircle) is an efficient diagnostic tool for L. infantum. Methods: The presence of L. infantum DNA was assessed in blood samples of 69 stray dogs using the conventional PCR (cPCR) and quantitative PCR (qPCR). Additional 50 lymph nodes and 50 bone marrow samples (positive and negative samples for parasitological tests) from dogs from endemic and nonendemic areas for CVL were also used. Results: L. infantum strains, and all positive lymph node and bone marrow samples for parasitological test gave positive results for cPCR and qPCR, presenting analytical sensitivity of ∼10° parasite mL−1. For the blood samples, 40/69 (58%; CI 95%; 46%-69%) resulted positive for L. infantum in both tests. All positive samples were confirmed by sequencing. Conclusion: This study showed the importance of the specific detection of L. infantum based on species-specific primers by molecular techniques, highlighting the application as a confirmation method in epidemiological studies and to adopt the best control measures.",

keywords = "Canine, KDNA, Leishmania infantum, PCR, Stray dogs",

author = "Silva, {Rodrigo C.} and Richini-Pereira, {Virg{\'i}nia B.} and Mariana Kikuti and Marson, {P{\^a}mela M.} and Helio Langoni",

note = "Funding Information: We thank Oswaldo Cruz Foundation, Leishmania Collection of the Oswaldo Cruz Institute (CLIOC), and Trypanosoma Collection of Wild and Domestic Mammals, and Vectors (ColTryp) for providing all 10 panels of Leishmania spp., and Trypanosoma cruzi strains, and S?o Paulo State University (UNESP) for the logistical support. Publisher Copyright: {\textcopyright} 2016 Informa UK Limited, trading as Taylor & Francis Group.",

year = "2017",

month = nov,

day = "1",

doi = "10.1080/01652176.2016.1252073",

language = "English (US)",

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T1 - Detection of Leishmania (L.) infantum in stray dogs by molecular techniques with sensitive species-specific primers

AU - Silva, Rodrigo C.

AU - Richini-Pereira, Virgínia B.

AU - Kikuti, Mariana

AU - Marson, Pâmela M.

AU - Langoni, Helio

N1 - Funding Information: We thank Oswaldo Cruz Foundation, Leishmania Collection of the Oswaldo Cruz Institute (CLIOC), and Trypanosoma Collection of Wild and Domestic Mammals, and Vectors (ColTryp) for providing all 10 panels of Leishmania spp., and Trypanosoma cruzi strains, and S?o Paulo State University (UNESP) for the logistical support. Publisher Copyright: © 2016 Informa UK Limited, trading as Taylor & Francis Group.

PY - 2017/11/1

Y1 - 2017/11/1

N2 - Background: Canine visceral leishmaniasis (CVL) is a worldwide parasitic zoonosis caused by Leishmania (Leishmania) infantum around the world. Canids are the definitive hosts and sand flies the intermediate hosts. Objective: To test the hypothesis that a new species-specific primers (Lch14:Lch15, targeting a multiple alignment for L. infantum kDNA minicircle) is an efficient diagnostic tool for L. infantum. Methods: The presence of L. infantum DNA was assessed in blood samples of 69 stray dogs using the conventional PCR (cPCR) and quantitative PCR (qPCR). Additional 50 lymph nodes and 50 bone marrow samples (positive and negative samples for parasitological tests) from dogs from endemic and nonendemic areas for CVL were also used. Results: L. infantum strains, and all positive lymph node and bone marrow samples for parasitological test gave positive results for cPCR and qPCR, presenting analytical sensitivity of ∼10° parasite mL−1. For the blood samples, 40/69 (58%; CI 95%; 46%-69%) resulted positive for L. infantum in both tests. All positive samples were confirmed by sequencing. Conclusion: This study showed the importance of the specific detection of L. infantum based on species-specific primers by molecular techniques, highlighting the application as a confirmation method in epidemiological studies and to adopt the best control measures.

AB - Background: Canine visceral leishmaniasis (CVL) is a worldwide parasitic zoonosis caused by Leishmania (Leishmania) infantum around the world. Canids are the definitive hosts and sand flies the intermediate hosts. Objective: To test the hypothesis that a new species-specific primers (Lch14:Lch15, targeting a multiple alignment for L. infantum kDNA minicircle) is an efficient diagnostic tool for L. infantum. Methods: The presence of L. infantum DNA was assessed in blood samples of 69 stray dogs using the conventional PCR (cPCR) and quantitative PCR (qPCR). Additional 50 lymph nodes and 50 bone marrow samples (positive and negative samples for parasitological tests) from dogs from endemic and nonendemic areas for CVL were also used. Results: L. infantum strains, and all positive lymph node and bone marrow samples for parasitological test gave positive results for cPCR and qPCR, presenting analytical sensitivity of ∼10° parasite mL−1. For the blood samples, 40/69 (58%; CI 95%; 46%-69%) resulted positive for L. infantum in both tests. All positive samples were confirmed by sequencing. Conclusion: This study showed the importance of the specific detection of L. infantum based on species-specific primers by molecular techniques, highlighting the application as a confirmation method in epidemiological studies and to adopt the best control measures.

KW - Canine

KW - KDNA

KW - Leishmania infantum

KW - PCR

KW - Stray dogs

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M3 - Article

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AN - SCOPUS:84994827752

SN - 0165-2176

VL - 37

SP - 23

EP - 30

JO - Veterinary Quarterly

JF - Veterinary Quarterly

IS - 1

ER -

Detection of Leishmania (L.) infantum in stray dogs by molecular techniques with sensitive species-specific primers

Abstract

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