@article{fdi:010079003,
  title = {{N}ew molecular approach for the detection of {K}inetoplastida parasites of medical and veterinary interest},
  author = {{M}edkour, {H}. and {V}arloud, {M}. and {D}avoust, {B}. and {M}ediannikov, {O}leg},
  editor = {},
  language = {{ENG}},
  abstract = {{K}inetoplastids are protozoa containing a range of ubiquitous free_living species-pathogens of invertebrates, vertebrates and even some plants. {S}ome of them are causative agents of canine vector-borne diseases. {T}heir diagnosis is often missing in a gold standard. {H}ere, we proposed a molecular approach for the diagnosis and study of {K}inetoplastida. {T}he {T}aq{M}an q{PCR} assays target the following genes: 24{S}a {LSU} of {K}inetoplastida, 28{S} {LSU} of {L}eishmania/{T}rypanosoma spp., 5.8{S} r{RNA} of {T}rypanosoma spp., 18{S} {SSU} of {L}eishmania spp., kinetoplast minicircle {DNA} (k{DNA}) of {L}. donovani complex and k{DNA} of {L}. infantum, were designed, validated for their sensitivity ({S}e) and specificity ({S}p) in silico and in vitro using a panel of known {DNA}s. {T}hey were then used to screen 369 blood samples (358 dogs, 2 equids, 9 monkeys). {I}n addition, new 28{S} {LSU} primer sets are presented to use for {K}inetoplastida's identification by {PCR}/sequencing. {A}ll q{PCR}s showed consistently high analytical sensitivities and reproducibility. {T}hey detect approximately 0.01 parasite/m{L} blood for the k{DNA} based-q{PCR}s and at least a single cell-equivalent of r{DNA} for the other systems. {B}ased on the sequencing results, after screening, {S}e and {S}p were: 0. 919 and 0.971, 0.853 and 0.979, 1.00 and 0.987, 0.826 and 0.995 for all of {K}inetoplastida, {L}eishmania/{T}rypanosoma, {T}rypanosoma, {L}eishmania spp. specific q{PCR}s, respectively. k{DNA} based q{PCR}s were more sensitive and specific ({S}e: 1.00; {S}p: 0.997). {PCR}/sequencing allowed the detection of {K}inetoplastids in animal blood samples such as {L}. infantum, {L}. guyanensis, {T}. congolense, {T}. evansi and {B}odo spp. {T}he molecular approach proposed here is useful for epidemiological studies, fundamental research such as screening for new {K}inetoplastida species, diagnosis and therapeutic follow-up. {I}n addition, researchers are free to choose the molecular tools adapted to their aims.},
  keywords = {{K}inetoplastida ; diagnostic ; q{PCR} ; {PCR} ; {L}eishmania ; {T}rypanosoma},
  booktitle = {},
  journal = {{M}icroorganisms},
  volume = {8},
  numero = {3},
  pages = {art. 356 [17 p.]},
  year = {2020},
  DOI = {10.3390/microorganisms8030356},
  URL = {https://www.documentation.ird.fr/hor/fdi:010079003},
}