Detection of Leishmania infantum kinetoplast minicircle DNA by Real Time PCR in hair of dogs with leishmaniosis

Belinchón-Lorenzo, Silvia; Iniesta, Virginia; Parejo, Juan Carlos; Fernández-Cotrina, Javier; Muñoz-Madrid, Rubén; Soto, Manuel; Alonso, Carlos; Nieto, Luis Carlos Gómez

doi:10.1016/j.vetpar.2012.11.007

Cited by 46 publications

(25 citation statements)

References 45 publications

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“…The use of qPCR provides the ability to perform very sensitive, accurate, and reproducible measurements of specific DNA present in a sample (Mortarino et al, 2004;Francino et al, 2006). Its high sensitivity and specificity have been reported using many different tissue types, including blood, lymph node, and bone marrow (Rodríguez-Cortés et al, 2007;Belinchón-Lorenzo et al, 2013). Regardless of the clinical form, we found that qPCR yielded the highest positivity for detecting CVL infection.…”

Section: Discussionmentioning

confidence: 54%

Molecular diagnosis of canine visceral leishmaniasis: A comparative study of three methods using skin and spleen from dogs with natural Leishmania infantum infection

Reis

Coura‐Vital

Roatt

et al. 2013

Veterinary Parasitology

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Polymerase chain reaction (PCR) and its variations represent highly sensitive and specific methods for Leishmania DNA detection and subsequent canine visceral leishmaniasis (CVL) diagnosis. The aim of this work was to compare three different molecular diagnosis techniques (conventional PCR [cPCR], seminested PCR [snPCR], and quantitative PCR [qPCR]) in samples of skin and spleen from 60 seropositive dogs by immunofluorescence antibody test and enzyme-linked immunosorbent assay. Parasitological analysis was conducted by culture of bone marrow aspirate and optical microscopic assessment of ear skin and spleen samples stained with Giemsa, the standard tests for CVL diagnosis. The primers L150/L152 and LINR4/LIN17/LIN19 were used to amplify the conserved region of the Leishmania kDNA minicircle in the cPCR, and snPCR and qPCR were performed using the DNA polymerase gene (DNA pol α) primers from Leishmania infantum. The parasitological analysis revealed parasites in 61.7% of the samples. Sensitivities were 89.2%, 86.5%, and 97.3% in the skin and 81.1%, 94.6%, and 100.0% in spleen samples used for cPCR, snPCR, and qPCR, respectively. We demonstrated that the qPCR method was the best technique to detect L. infantum in both skin and spleen samples. However, we recommend the use of skin due to the high sensitivity and sampling being less invasive.

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Section: Discussionmentioning

confidence: 54%

Molecular diagnosis of canine visceral leishmaniasis: A comparative study of three methods using skin and spleen from dogs with natural Leishmania infantum infection

Reis

Coura‐Vital

Roatt

et al. 2013

Veterinary Parasitology

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“…Analysis of dog hair is an optimal and noninvasive method for diagnosis and monitoring treatment response in canine leishmaniasis when molecular techniques are used (Corpas‐López et al., ) and may be very useful in epidemiological studies (Belinchón‐Lorenzo et al., ). In El Borouj, L. tropica was detected in 3.0% of the dogs through the PCR analysis of their hair.…”

Section: Discussionmentioning

confidence: 99%

Risk factors for the expansion of cutaneous leishmaniasis by Leishmania tropica : Possible implications for control programmes

Gijón-Robles

Abattouy

Merino‐Espinosa

et al. 2018

Transbound Emerg Dis

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Cutaneous leishmaniasis (CL) caused by Leishmania tropica is emerging in new areas, initially as outbreaks and then establishing endemic foci. There is little evidence of the risk factors and effectiveness of existing control measures, what limits our ability to generalize in different epidemiological settings. The disease is described as anthroponotic; however, zoonotic outbreaks have been reported in some countries. Our aim was to identify risk factors in a recently reported endemic focus in Morocco in order to design more effective control programmes. A case-control study was conducted from September 2014 to October 2015 for epidemiological data collection from families with and without CL cases. Sandflies were captured and L. tropica infection determined. The presence of potential animal reservoirs was evaluated. 71 CL cases (44 diagnosed between 2013 and 2015) and 137 healthy people were surveyed. The average age of the new cases was 33.1 ± 22.3 years, and 69.0% were women. Phlebotomus sergenti was the most abundant species with a density of 4.27 sandflies/trap/night and differences between houses with and without CL cases were detected (p-value = 0.014). Overall, 2.7% female P. sergenti and 3.0% dogs were positive for L. tropica. Human, cat, rabbit and bird blood was detected in blood-fed P. sergenti females. 45% people used preventive measures that were not translated into a reduction in the individual risk of acquiring CL. Exposure to P. sergenti was the only risk factor found, and the reduction in its density could be achieved through the improvement of water wells management, organic fertilizers' disposal and dogs control. The lack of effectiveness of indoor residual spraying and treated nets are attributable to poor compliance and misuse of them. In addition, result optimization of the awareness campaigns on the public is possible by involving patients with CL to explain their own experience.

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“…Similarly, Ferreira et al (2013), using qPCR, obtained positivity values on the order of 90% for blood samples, 97% for skin biopsy, 98% for conjunctival swab samples and 100% for bone marrow samples [134]. Other studies showed better results for sensitivity to certain tissue, such as Maya et al (2009) that evaluated dogs with different clinical profiles and demonstrated that the use of cPCR for parasite DNA detection on lymph node aspirate would be ideal for the early CVL diagnosis in symptomatic animals [132], a finding later corroborated by Lombardo et al (2012) and Belinchon-Lorenzo (2013) [135,136]. However, in the absence of lymphadenopathy, other studies showed that bone marrow aspirates offered better sensitivity, since it has a higher parasite load in relation to lymph nodes [132,137].…”

Section: Pcrmentioning

confidence: 93%

“…Recently, Belinchon-Lorenzo (2013), using qPCR, demonstrated the presence of Leishmania kDNA in the hair and keratinocytes of infected animals. According to the authors, the use of the non-invasive hair sample for the diagnosis of CVL would be advantageous because it is easy to collect, handle, transport, and store [135]. However, further studies should be conducted to determine the sensitivity of this method.…”

Section: Pcrmentioning

confidence: 99%

New Advances in the Diagnosis of Canine Visceral Leishmaniasis

Veras¹,

Solcà²,

Guedes³

2014

Leishmaniasis - Trends in Epidemiology, Diagnosis and Treatment

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Detection of Leishmania infantum kinetoplast minicircle DNA by Real Time PCR in hair of dogs with leishmaniosis

Cited by 46 publications

References 45 publications

Molecular diagnosis of canine visceral leishmaniasis: A comparative study of three methods using skin and spleen from dogs with natural Leishmania infantum infection

Molecular diagnosis of canine visceral leishmaniasis: A comparative study of three methods using skin and spleen from dogs with natural Leishmania infantum infection

Risk factors for the expansion of cutaneous leishmaniasis by Leishmania tropica : Possible implications for control programmes

New Advances in the Diagnosis of Canine Visceral Leishmaniasis

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