Detection and chronology of parasitic kinetoplast DNA presence in hair of experimental Leishmania major infected BALB/c mice by Real Time PCR
The present work describes a new sensitive qPCR method for Leishmania major kDNA detection in hair samples from experimental infected BALB/c mice. •Hair samples from Leishmania major experimental infected BALB/c mice were analyzed by qPCR.•Hairs were collected and analyzed weekly for 35 days after c...
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Veröffentlicht in: | Acta tropica 2013-12, Vol.128 (3), p.468-472 |
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Sprache: | eng |
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Zusammenfassung: | The present work describes a new sensitive qPCR method for Leishmania major kDNA detection in hair samples from experimental infected BALB/c mice.
•Hair samples from Leishmania major experimental infected BALB/c mice were analyzed by qPCR.•Hairs were collected and analyzed weekly for 35 days after challenge.•Samples proceeded both from ear (site of inoculation) and leg (healthy skin).•Parasite kDNA was detected in all ear and leg hair samples at the final timepoint.•Hair is a suitable sample for the monitoring of L. major infection.
Hair can accumulate foreign chemical or biological substances. Recently, it has been reported that parasite DNA can also be detected in the hair of Leishmania infantum infected dogs. The aim of this work has been to find out whether parasite DNA incorporates in the hair of Leishmania major experimentally infected animals. For this purpose, a group of 4 BALB/c mice, intradermally inoculated in both ears with 1000 L. major V1 strain promastigote forms, was monitored for parameters associated to the infection during 35 days. Weekly, ear swelling was measured, and hair samples from ears and leg were collected. Blood samples were obtained before challenge and at day 35 post infection, when parasite load was measured in ear, lymph node and spleen by limit dilution. Ear swelling and other parameters observed in the infected mice were consistent with those described for this model. The presence of parasite kinetoplast DNA (kDNA) was detected by Real Time PCR in all ear and leg hair samples at the final timepoint. These data suggests that hair is a specialized tissue in the sequestration and removal of foreign DNA. Detection of DNA in hair could be, therefore, a useful tool to chronologically record the infection process during experimental mice assays. |
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ISSN: | 0001-706X 1873-6254 |
DOI: | 10.1016/j.actatropica.2013.07.007 |