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Use of transfer RNA-intergenic spacer PCR combined with capillary electrophoresis to identify coagulase-negative Staphylococcus species originating from bovine milk and teat apices

Karlien Supré UGent, 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 (2009) JOURNAL OF DAIRY SCIENCE. 92(7). p.3204-3210
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.
Please use this url to cite or link to this publication:
author
organization
alternative title
Technical note: Use of transfer RNA-intergenic spacer PCR combined with capillary electrophoresis to identify coagulase-negative Staphylococcus species originating from bovine milk and teat apices
year
type
journalArticle (original)
publication status
published
subject
keyword
IDENTIFICATION, LENGTH POLYMORPHISM ANALYSIS, transfer RNA-intergenic spacer polymerase chain reaction, teat apex, bovine coagulase-negative staphylococci, REGION, NOV., milk
journal title
JOURNAL OF DAIRY SCIENCE
J. Dairy Sci.
volume
92
issue
7
pages
3204 - 3210
Web of Science type
Article
Web of Science id
000267022500023
JCR category
AGRICULTURE, DAIRY & ANIMAL SCIENCE
JCR impact factor
2.463 (2009)
JCR rank
2/49 (2009)
JCR quartile
1 (2009)
ISSN
0022-0302
DOI
10.3168/jds.2008-1923
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
676958
handle
http://hdl.handle.net/1854/LU-676958
date created
2009-06-04 08:28:48
date last changed
2012-10-31 09:00:04
@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       = {Supr{\'e}, 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},
  keyword      = {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://dx.doi.org/10.3168/jds.2008-1923},
  volume       = {92},
  year         = {2009},
}

Chicago
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.
APA
Supré, K., De Vliegher, S., Sampimon, O. C., Zadoks, R. N., Vaneechoutte, M., Baele, M., De Graef, E., et al. (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.
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.
MLA
Supré, Karlien, Sarne De Vliegher, Otlis C Sampimon, 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 92.7 (2009): 3204–3210. Print.