Publication: Sensitivity of two methods to detect Mycoplasma agalactiae in goat milk
Authors
Tatay-Dualde, Juan ; Prats- Van der Ham, Miranda ; Gómez-Martín, Angel ; Paterna, Ana ; Corrales, Juan Carlos ; Sánchez, Antonio ; Fe, Christian de la ; Amores, Joaquín
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Amores, Joaquín
Publisher
BioMed Central
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DOI
10.1186/s13620-015-0049-y
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info:eu-repo/semantics/article
Description
© 2015 Tatay-Dualde et al. Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver
(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Abstract
Background: Laboratory diagnostic techniques able to detect Mycoplasma agalactiae are essential in contagious
agalactia in dairy goats. This study was designed: 1) to determine the detection limits of PCR and culture in goat
milk samples, 2) to examine the effects of experimental conditions including the DNA extraction method, PCR
technique and storage conditions (fresh versus frozen stored milk samples) on these methods and 3), to establish
agreement between PCR and culture techniques using milk samples from goats with mastitis in commercial dairy
herds. The study was conducted both on artificially inoculated and field samples.
Results: Our findings indicate that culture is able to detect M. agalactiae in goat milk at lower concentrations than
PCR. Qualitative detection of M.agalactiae by culture and PCR was not affected by sample freezing, though the
DNA extraction method used significantly affected the results of the different PCR protocols. When clinical samples
were used, both techniques showed good agreement.
Conclusions: The results from this study indicate that both culture and PCR are able to detect M. agalactiae in clinical
goat mastitis samples. However, in bulk tank milk samples with presumably lower M. agalactiae concentrations, culture
is recommended within the first 24 h of sample collection due to its lower limit of detection. To improve the diagnostic
sensitivity of PCR in milk samples, there is a need to increase the efficiency of extracting DNA from milk samples using
protocols including a previous step of enzymatic digestion.
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Mycoplasma , Sensibilidad , Goat , Contagious agalactia , PCR , Culture , Milk , Mycoplasma agalactiae
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