Use of transfer RNA-intergenic spacer PCR combined with capillary electrophoresis to identify coagulase-negative Staphylococcus species originating from bovine milk and teat apices
- Author
- Karlien Supré, Sarne De Vliegher (UGent) , Otlis C Sampimon, Ruth N Zadoks, Mario Vaneechoutte (UGent) , Margo Baele (UGent) , Evelyne De Graef (UGent) , Sofie Piepers (UGent) and Freddy Haesebrouck (UGent)
- Organization
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- Department of Clinical chemistry, microbiology and immunology (ceased 1-10-2018)
- Department of Pathology, bacteriology and poultry diseases (ceased 1-1-2022)
- Department of Obstetrics, reproduction and herd health (ceased 1-1-2022)
- Department of Internal Medicine, Reproduction and Population Medicine
- Abstract
- Coagulase-negative staphylococci (CNS) are the most frequently isolated bacteria in milk samples from cows with and without mastitis. Elucidating their relevance in bovine udder health is hampered because identification at species level, if done at all, used to be performed based on phenotypic features. In order to provide a rapid, cheap, and easy-to-use genotypic technique that can be used to identify CNS species from milk and teat apices from cows, the performance of tRNA-intergenic spacer PCR (tDNA-PCR) in combination with capillary electrophoresis was evaluated. After updating the tDNA library with CNS reference strains, 288 field isolates were identified with tDNA-PCR and gene sequencing, and the latter was used as the reference method. The field isolates were divided in 2 groups of 144. Isolates of the first group were identified with tDNA-PCR with a typeability of 81.9% and an accuracy of 94.1%. Peak patterns of these isolates were then added to the tDNA library with species identity as determined by DNA sequencing. The second group was identified with the updated tDNA library, resulting in 91.0% typeability and 99.2% accuracy. This study showed that the updated tDNA-PCR in combination with capillary electrophoresis was almost as accurate as gene sequencing but faster and cheaper (only $3 per isolate), and is a useful tool in observational studies concerning the epidemiology of bovine CNS species.
- Keywords
- IDENTIFICATION, LENGTH POLYMORPHISM ANALYSIS, transfer RNA-intergenic spacer polymerase chain reaction, teat apex, bovine coagulase-negative staphylococci, REGION, NOV., milk
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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-676958
- MLA
- Supré, Karlien, et al. “Use of Transfer RNA-Intergenic Spacer PCR Combined with Capillary Electrophoresis to Identify Coagulase-Negative Staphylococcus Species Originating from Bovine Milk and Teat Apices.” JOURNAL OF DAIRY SCIENCE, vol. 92, no. 7, 2009, pp. 3204–10, doi:10.3168/jds.2008-1923.
- APA
- Supré, K., De Vliegher, S., Sampimon, O. C., Zadoks, R. N., Vaneechoutte, M., Baele, M., … Haesebrouck, F. (2009). Use of transfer RNA-intergenic spacer PCR combined with capillary electrophoresis to identify coagulase-negative Staphylococcus species originating from bovine milk and teat apices. JOURNAL OF DAIRY SCIENCE, 92(7), 3204–3210. https://doi.org/10.3168/jds.2008-1923
- Chicago author-date
- Supré, Karlien, Sarne De Vliegher, Otlis C Sampimon, Ruth N Zadoks, Mario Vaneechoutte, Margo Baele, Evelyne De Graef, Sofie Piepers, and Freddy Haesebrouck. 2009. “Use of Transfer RNA-Intergenic Spacer PCR Combined with Capillary Electrophoresis to Identify Coagulase-Negative Staphylococcus Species Originating from Bovine Milk and Teat Apices.” JOURNAL OF DAIRY SCIENCE 92 (7): 3204–10. https://doi.org/10.3168/jds.2008-1923.
- Chicago author-date (all authors)
- Supré, Karlien, Sarne De Vliegher, Otlis C Sampimon, Ruth N Zadoks, Mario Vaneechoutte, Margo Baele, Evelyne De Graef, Sofie Piepers, and Freddy Haesebrouck. 2009. “Use of Transfer RNA-Intergenic Spacer PCR Combined with Capillary Electrophoresis to Identify Coagulase-Negative Staphylococcus Species Originating from Bovine Milk and Teat Apices.” JOURNAL OF DAIRY SCIENCE 92 (7): 3204–3210. doi:10.3168/jds.2008-1923.
- Vancouver
- 1.Supré K, De Vliegher S, Sampimon OC, Zadoks RN, Vaneechoutte M, Baele M, et al. Use of transfer RNA-intergenic spacer PCR combined with capillary electrophoresis to identify coagulase-negative Staphylococcus species originating from bovine milk and teat apices. JOURNAL OF DAIRY SCIENCE. 2009;92(7):3204–10.
- IEEE
- [1]K. Supré et al., “Use of transfer RNA-intergenic spacer PCR combined with capillary electrophoresis to identify coagulase-negative Staphylococcus species originating from bovine milk and teat apices,” JOURNAL OF DAIRY SCIENCE, vol. 92, no. 7, pp. 3204–3210, 2009.
@article{676958, abstract = {{Coagulase-negative staphylococci (CNS) are the most frequently isolated bacteria in milk samples from cows with and without mastitis. Elucidating their relevance in bovine udder health is hampered because identification at species level, if done at all, used to be performed based on phenotypic features. In order to provide a rapid, cheap, and easy-to-use genotypic technique that can be used to identify CNS species from milk and teat apices from cows, the performance of tRNA-intergenic spacer PCR (tDNA-PCR) in combination with capillary electrophoresis was evaluated. After updating the tDNA library with CNS reference strains, 288 field isolates were identified with tDNA-PCR and gene sequencing, and the latter was used as the reference method. The field isolates were divided in 2 groups of 144. Isolates of the first group were identified with tDNA-PCR with a typeability of 81.9% and an accuracy of 94.1%. Peak patterns of these isolates were then added to the tDNA library with species identity as determined by DNA sequencing. The second group was identified with the updated tDNA library, resulting in 91.0% typeability and 99.2% accuracy. This study showed that the updated tDNA-PCR in combination with capillary electrophoresis was almost as accurate as gene sequencing but faster and cheaper (only $3 per isolate), and is a useful tool in observational studies concerning the epidemiology of bovine CNS species.}}, author = {{Supré, Karlien and De Vliegher, Sarne and Sampimon, Otlis C and Zadoks, Ruth N and Vaneechoutte, Mario and Baele, Margo and De Graef, Evelyne and Piepers, Sofie and Haesebrouck, Freddy}}, issn = {{0022-0302}}, journal = {{JOURNAL OF DAIRY SCIENCE}}, keywords = {{IDENTIFICATION,LENGTH POLYMORPHISM ANALYSIS,transfer RNA-intergenic spacer polymerase chain reaction,teat apex,bovine coagulase-negative staphylococci,REGION,NOV.,milk}}, language = {{eng}}, number = {{7}}, pages = {{3204--3210}}, title = {{Use of transfer RNA-intergenic spacer PCR combined with capillary electrophoresis to identify coagulase-negative Staphylococcus species originating from bovine milk and teat apices}}, url = {{http://doi.org/10.3168/jds.2008-1923}}, volume = {{92}}, year = {{2009}}, }
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