Literaturdatenbank
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duPre, S. A., Tracy, C. R., & Hunter, K. W. (2011). Quantitative pcr method for detection of mycoplasma spp. dna in nasal lavage samples from the desert tortoise (gopherus agassizii). Journal of Microbiological Methods, (Article in Press, Uncorrected Proof).
Added by: Sarina Wunderlich (25 Jun 2011 12:41:27 UTC) |
| Resource type: Journal Article BibTeX citation key: duPre2011 View all bibliographic details  |
Categories: General Keywords: Bakterien = bacteria, Gopherus, Gopherus agassizii, Schildkröten = turtles + tortoises, Testudinidae, Veterinärmedizin = veterinary medicine Creators: duPre, Hunter, Tracy Collection: Journal of Microbiological Methods |
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| Abstract |
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Testudinidae Mycoplasma agassizii and M. testudineum have been associated with upper respiratory tract disease (URTD) in the threatened desert tortoise (Gopherus agassizii). Because microbiological culture methods have proven difficult to employ in wild desert tortoises, our goal was to develop a sensitive and specific qPCR method for detecting and quantifying mycoplasma DNA in nasal lavage fluid collected in the field. Primers for 16S ribosomal RNA gene sequences specific for M. agassizii and M. testudineum were designed, together with primers that recognize conserved sequences of both microorganisms. Standard curves generated with DNA extracted from known numbers of mycoplasma cells revealed a lower detection limit of approximately 5 fg. The qPCR method did not recognize normal flora DNA, and nasal lavage fluid contained no interfering substances. Nasal lavage samples collected from 20 captive desert tortoises housed at the Desert Tortoise Conservation Center (Clark County, Nevada, USA) revealed the presence of M. agassizii DNA in 100% of the tortoises. Concentrations ranged from a low of 6 pg ml- 1 to a high of 72,962 pg ml- 1. Only one of the tortoises was positive for M. testudineum. Interestingly, not all of the qPCR positive tortoises showed evidence of seroconversion, suggesting that they were colonized but not infected. This new quantitative method will provide a critical tool for managing threatened populations of the desert tortoise.
Added by: Sarina Wunderlich |