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Article

New Molecular Approach for the Detection of Kinetoplastida Parasites of Medical and Veterinary Interest

1
IHU Méditerranée Infection - Microbes, Evolution, Phylogeny and Infection (MEΦI), 13385 Marseille CEDEX 05, France
2
UMR Aix-Marseille Université, IRD, APHM -19-21, Bd Jean Moulin, 13385 Marseille CEDEX 05, France
3
PADESCA Laboratory, Veterinary Science Institute, University Constantine 1, El Khroub 25100, Algeria
4
CEVA Animal Health, 33500 Libourne, France
*
Author to whom correspondence should be addressed.
Microorganisms 2020, 8(3), 356; https://doi.org/10.3390/microorganisms8030356
Received: 15 February 2020 / Revised: 26 February 2020 / Accepted: 1 March 2020 / Published: 2 March 2020
(This article belongs to the Special Issue Canine Vector Borne Diseases)
Kinetoplastids are protozoa containing a range of ubiquitous free_living species–pathogens of invertebrates, vertebrates and even some plants. Some of them are causative agents of canine vector-borne diseases. Their diagnosis is often missing in a gold standard. Here, we proposed a molecular approach for the diagnosis and study of Kinetoplastida. The TaqMan qPCR assays target the following genes: 24Sa LSU of Kinetoplastida, 28S LSU of Leishmania/ Trypanosoma spp., 5.8S rRNA of Trypanosoma spp., 18S SSU of Leishmania spp., kinetoplast minicircle DNA (kDNA) of L. donovani complex and kDNA of L. infantum, were designed, validated for their sensitivity (Se) and specificity (Sp) in silico and in vitro using a panel of known DNAs. They were then used to screen 369 blood samples (358 dogs, 2 equids, 9 monkeys). In addition, new 28S LSU primer sets are presented to use for Kinetoplastida’s identification by PCR/sequencing. All qPCRs showed consistently high analytical sensitivities and reproducibility. They detect approximately 0.01 parasite/ mL blood for the kDNA based- qPCRs and at least a single cell-equivalent of rDNA for the other systems. Based on the sequencing results, after screening, Se and Sp were: 0. 919 and 0.971, 0.853 and 0.979, 1.00 and 0.987, 0.826 and 0.995 for all of Kinetoplastida, Leishmania/ Trypanosoma, Trypanosoma, Leishmania spp. specific qPCRs, respectively. kDNA based qPCRs were more sensitive and specific (Se: 1.00; Sp: 0.997). PCR/sequencing allowed the detection of Kinetoplastids in animal blood samples such as L. infantum, L. guyanensis, T. congolense, T. evansi and Bodo spp. The molecular approach proposed here is useful for epidemiological studies, fundamental research such as screening for new Kinetoplastida species, diagnosis and therapeutic follow-up. In addition, researchers are free to choose the molecular tools adapted to their aims. View Full-Text
Keywords: Kinetoplastida; diagnostic; qPCR; PCR; Leishmania; Trypanosoma Kinetoplastida; diagnostic; qPCR; PCR; Leishmania; Trypanosoma
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MDPI and ACS Style

Medkour, H.; Varloud, M.; Davoust, B.; Mediannikov, O. New Molecular Approach for the Detection of Kinetoplastida Parasites of Medical and Veterinary Interest. Microorganisms 2020, 8, 356. https://doi.org/10.3390/microorganisms8030356

AMA Style

Medkour H, Varloud M, Davoust B, Mediannikov O. New Molecular Approach for the Detection of Kinetoplastida Parasites of Medical and Veterinary Interest. Microorganisms. 2020; 8(3):356. https://doi.org/10.3390/microorganisms8030356

Chicago/Turabian Style

Medkour, Hacène, Marie Varloud, Bernard Davoust, and Oleg Mediannikov. 2020. "New Molecular Approach for the Detection of Kinetoplastida Parasites of Medical and Veterinary Interest" Microorganisms 8, no. 3: 356. https://doi.org/10.3390/microorganisms8030356

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