First detection of Leishmania kDNA in canine cerumen samples by qPCR

Belinchón-Lorenzo, Silvia; Parejo, Juan Carlos; Iniesta, Virginia; Fernández-Cotrina, Javier; Muñoz-Madrid, Rubén; Monroy, Isabel; Baz, Victoria; Gómez-Luque, Adela; Serrano-Aguilera, Francisco Javier; Barneto, José Luis; Gómez-Nieto, Luis Carlos

doi:10.1016/j.vetpar.2016.05.021

Cited by 12 publications

(8 citation statements)

References 23 publications

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“…PCR reactions were carried out in 96 wells PCR plates in a final volume of 20 microlitres (4 microlitres of DNA + 16 microlitres of Reaction Mix), containing 20 microM of each primer, 10 microM of TaqMan Probe and the iTaq Universal Probes Supermix f . Primers, probe and the thermal cycling profile used are described elsewhere 26 . Each amplification run contained positive and negative controls.…”

Section: Methodsmentioning

confidence: 99%

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Association of myocardial parasitic load with cardiac biomarkers and other selected variables in 10 dogs with advanced Canine Leishmaniasis

et al. 2021

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Background: The association between myocardial parasitic load (MPL) and cardiac biomarkers in Canine Leishmaniasis (CanL) has not been studied. Methods: Dogs with advanced CanL were prospectively recruited and were included if they were euthanised. Prior to euthanasia these variables were assessed: hematocrit, globulin, creatinine, N‐terminal‐pro brain natriuretic peptide (NT‐proBNP), cardiac troponin I (cTnI), blood pressure, urine protein/creatinine ratio and echocardiographic parameters. A left ventricular (LV) sample was taken for histopathology and MPL evaluation by quantitative PCR. Correlation of MPL with all variables was analysed. Dogs with lower and higher histopathology scores were compared. Results: Ten dogs were included. NT‐proBNP was 6946 pmol/ (interquartile range [IQR] 3751–9268 pmol/L) and cTnI 4.56 ng/mL (IQR 0.46–13.1 ng/mL). In all dogs, echocardiography showed an increase in LV thickening, and histopathology revealed moderate to severe lympho‐plasmocytic myocarditis and/or myocardial cell degeneration. MPL was 215.53 parasites/gram (IQR 21.2–1372.63 parasites/gram). A strong correlation (p < 0.001; R = 0.90; R20.81) with cTnI was observed but correlation with any of the other variables or differences between the two histopathological scores, were not detected. Conclusions: MPL in dogs with advanced CanL shows variable but generally high levels. A strong association between MPL and cTnI was observed, which encourages the exploration of cTnI as a marker in CanL.

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Section: Methodsmentioning

confidence: 99%

“…Primers, probe and the thermal cycling profile used are described elsewhere. 26 Each amplification run contained positive and negative controls. All qPCR analyses were performed in a Step One Plus Real Time PCR System g .…”

Section: Methodsmentioning

confidence: 99%

Association of myocardial parasitic load with cardiac biomarkers and other selected variables in 10 dogs with advanced Canine Leishmaniasis

et al. 2021

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“…In relation to malaria diagnosis by qPCR, the presence of Plasmodium DNA in other samples besides blood, such as urine and saliva, was recently demonstrated [16][17][18][19]. The recent demonstration of foreign parasitic DNA presence in hair and epidermal keratinocytes of Leishmania-infected animals opened a new suitable scientific field to study the physiology of these epithelial cells during several diseases, acting as a very specialized tissue to sequestrate and eliminate foreign genetic material [20][21][22]36,37]. The detection of P. falciparum DNA in malaria patients' hair confirms again the excretion and purification function of this tissue.…”

Section: Discussionmentioning

confidence: 99%

Method for Malaria Diagnosis Based on Extractions of Samples Using Non-Invasive Techniques: An Opportunity for the Nursing Clinical Practice

Gómez-Luque

Parejo

Clavijo-Chamorro

et al. 2020

IJERPH

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Malaria has been for millennia one of the best known and most destructive diseases affecting humans. Its high impact has aroused great interest for the development of new effective and reliable diagnostic techniques. Recently it has been recently published that hairs from mammal hosts are able to capture, hold and finally remove foreign DNA sequences of Leishmania parasites. The aim of this study was to check if Plasmodium falciparum (P. falciparum) DNA remains stable in blood samples deposited in Whatman paper after suffering different transport and storage conditions, and to compare the sensitivity of these results with those offered by thick a smear and Rapid Diagnostic Test, and besides to examine whether P. falciparum DNA would be detected and quantified by Real time quantitative PCR (qPCR) from hairs of people with different types of malaria. P. falciparum Histidine Repeat Protein II (pHRP-II) antigen detection and P. falciparum DNA were detected in 18 of 19 dry blood samples adhered to Whatman paper (94.74%), besides, Plasmodium DNA was also detected in seven out of 19 hair samples analyzed (36.84%), remaining stable until analysis for several months under the exposure to different environmental conditions. Although the sensitivity of PCR for the diagnosis of malaria in hair samples is not as high as blood analysis, the study of Plasmodium DNA presence in blood and hair could constitute a complementary tool with numerous advantages in sample collection, transport and storage. We suggest that the method could be also applied to medical, forensic and paleo-parasitological diagnosis, not only for malaria but also for searching many other pathogens in hair samples.

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“…• Human and animal leishmaniases. Leishmania DNA has been detected and quantified in the cerumen of infected dogs [336]. A recent publication demonstrates that cerumen-qPCR expresses the highest sensitivity (87.5%) to detect genetic materials, followed by hair (lesions: 78.57%, healthy skin: 62.5%), and blood (68.75%) [337].…”

Section: Ear Swab/cerumenmentioning

confidence: 99%

Noninvasive Biological Samples to Detect and Diagnose Infections due to Trypanosomatidae Parasites: A Systematic Review and Meta-Analysis

Sereno¹,

Akhoundi²,

Sayehmri³

et al. 2020

Preprint

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Unicellular eukaryotes of the Trypanosomatidae family include human and animal pathogens that belong to the Trypanosoma and Leishmania genera. Diagnosis of the diseases they caused requires the sampling of body fluids (blood, lymph, peritoneal fluid, cerebrospinal fluid, etc.) or organ biopsies (bone marrow, spleen, etc.), which are mostly obtained through invasive methods. Body fluids or appendages can be alternatives to these invasive biopsies but appropriateness remains poorly studied. To further address this question, we perform a systematic review on clues evidencing the presence of parasites, genetic material, antibodies, and antigens in body secretions, appendages, or the organs or proximal tissues that produce these materials. Paper selection was based on searches in PubMed, Web of Science, WorldWideScience, SciELO, Embase, Google. The information of each selected article (n=333) was classified into different sections and data were extracted from 77 papers. The presence of Trypanosomatidae parasites has been tracked in most of organs or proximal tissues that produce body secretions or appendages, in naturally or experimentally infected hosts. The meta-analysis highlights the paucity of studies on Human African Trypanosomiasis and a the absence on animal Trypanosomiasis. Among the collected data high heterogeneity in terms of the I2 statistic (100%) is recorded. A high positivity is recorded for antibody and genetic material detection in urine of patients and dogs suffering leishmaniasis, and of antigen for leishmaniasis and Chagas disease. Data on conjunctival swab can be analyzed with molecular methods solely for dogs suffering canine visceral leishmaniasis. Saliva and hair/bristle showed a pretty good positivity that support their potential to be used for leishmaniasis diagnosis. In conclusion, our study pinpoints significant gaps that need to be filled in order to properly address the interest of body secretion and hair or bristle for the diagnosis of infections caused by Leishmania and by other Trypanosomatidae parasites.

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First detection of Leishmania kDNA in canine cerumen samples by qPCR

Cited by 12 publications

References 23 publications

Association of myocardial parasitic load with cardiac biomarkers and other selected variables in 10 dogs with advanced Canine Leishmaniasis

Association of myocardial parasitic load with cardiac biomarkers and other selected variables in 10 dogs with advanced Canine Leishmaniasis

Method for Malaria Diagnosis Based on Extractions of Samples Using Non-Invasive Techniques: An Opportunity for the Nursing Clinical Practice

Noninvasive Biological Samples to Detect and Diagnose Infections due to Trypanosomatidae Parasites: A Systematic Review and Meta-Analysis

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