Improved rapid and efficient method for Staphylococcus aureus DNA extraction from milk for identification of mastitis pathogens

A rapid and efficient DNA extraction method was developed for detecting mastitis pathogens in milk. The first critical step involved cell wall disruption by bead-beating, as physical disruption using beads was more effective for DNA extraction from Gram-positive bacteria, such as Staphylococcus aure...

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Published in	Journal of Veterinary Medical Science Vol. 77; no. 8; pp. 1007 - 1009
Main Authors	UNNO, Hirotaka, KIKU, Yoshio, NAKAMURA, Akiyoshi, INADA, Mika, TAGAWA, Yuichi, ITO, Keiko, HAYASHI, Tomohito, HATA, Eiji, HASHIMOTO, Michie, KATSUDA, Ken, NIKAIDO, Masaru, KAWAI, Kazuhiro, HASHIMOTO, Koji
Format	Journal Article
Language	English
Published	Japan JAPANESE SOCIETY OF VETERINARY SCIENCE 01.01.2015 Japan Science and Technology Agency The Japanese Society of Veterinary Science
Subjects	Animals Bacteriology Cattle cow milk DNA chip DNA extraction DNA, Bacterial - isolation & purification Female loop-mediated isothermal amplification (LAMP) mastitis Mastitis, Bovine - diagnosis Mastitis, Bovine - microbiology Milk - chemistry Milk - microbiology Nucleic Acid Amplification Techniques - veterinary pathogen Staphylococcal Infections - diagnosis Staphylococcal Infections - microbiology Staphylococcal Infections - veterinary Staphylococcus aureus Staphylococcus aureus - genetics
Online Access	Get full text
ISSN	0916-7250 1347-7439 1347-7439
DOI	10.1292/jvms.14-0159

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Abstract	A rapid and efficient DNA extraction method was developed for detecting mastitis pathogens in milk. The first critical step involved cell wall disruption by bead-beating, as physical disruption using beads was more effective for DNA extraction from Gram-positive bacteria, such as Staphylococcus aureus, than enzymatic disruption using proteinase K. The second critical step involves the use of acetic acid and ammonium sulfate in the purification process, as these reagents effectively and efficiently remove the lipids and proteins in milk. Using these methods, DNA suitable for loop-mediated isothermal amplification was obtained within 30 min. Also, the rapid and sensitive detection of S. aureus in milk was possible at levels as low as 200 cfu/ml.
AbstractList	A rapid and efficient DNA extraction method was developed for detecting mastitis pathogens in milk. The first critical step involved cell wall disruption by bead-beating, as physical disruption using beads was more effective for DNA extraction from Gram-positive bacteria, such as Staphylococcus aureus, than enzymatic disruption using proteinase K. The second critical step involves the use of acetic acid and ammonium sulfate in the purification process, as these reagents effectively and efficiently remove the lipids and proteins in milk. Using these methods, DNA suitable for loop-mediated isothermal amplification was obtained within 30 min. Also, the rapid and sensitive detection of S. aureus in milk was possible at levels as low as 200 cfu/ml. A rapid and efficient DNA extraction method was developed for detecting mastitis pathogens in milk. The first critical step involved cell wall disruption by bead-beating, as physical disruption using beads was more effective for DNA extraction from Gram-positive bacteria, such as Staphylococcus aureus, than enzymatic disruption using proteinase K. The second critical step involves the use of acetic acid and ammonium sulfate in the purification process, as these reagents effectively and efficiently remove the lipids and proteins in milk. Using these methods, DNA suitable for loop-mediated isothermal amplification was obtained within 30 min. Also, the rapid and sensitive detection of S. aureus in milk was possible at levels as low as 200 cfu/ml.A rapid and efficient DNA extraction method was developed for detecting mastitis pathogens in milk. The first critical step involved cell wall disruption by bead-beating, as physical disruption using beads was more effective for DNA extraction from Gram-positive bacteria, such as Staphylococcus aureus, than enzymatic disruption using proteinase K. The second critical step involves the use of acetic acid and ammonium sulfate in the purification process, as these reagents effectively and efficiently remove the lipids and proteins in milk. Using these methods, DNA suitable for loop-mediated isothermal amplification was obtained within 30 min. Also, the rapid and sensitive detection of S. aureus in milk was possible at levels as low as 200 cfu/ml. A rapid and efficient DNA extraction method was developed for detecting mastitis pathogens in milk. The first critical step involved cell wall disruption by bead-beating, as physical disruption using beads was more effective for DNA extraction from Gram-positive bacteria, such as Staphylococcus aureus , than enzymatic disruption using proteinase K. The second critical step involves the use of acetic acid and ammonium sulfate in the purification process, as these reagents effectively and efficiently remove the lipids and proteins in milk. Using these methods, DNA suitable for loop-mediated isothermal amplification was obtained within 30 min. Also, the rapid and sensitive detection of S. aureus in milk was possible at levels as low as 200 cfu/m l .
Author	UNNO, Hirotaka HASHIMOTO, Koji HAYASHI, Tomohito HATA, Eiji INADA, Mika NIKAIDO, Masaru KIKU, Yoshio NAKAMURA, Akiyoshi KAWAI, Kazuhiro HASHIMOTO, Michie TAGAWA, Yuichi ITO, Keiko KATSUDA, Ken
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References	4. Cha, E., Bar, D., Hertl, J. A., Tauer, L. W., Bennett, G., González, R. N., Schukken, Y. H., Welcome, F. L. and Gröhn, Y. T. 2011. The cost and management of different types of clinical mastitis in dairy cows estimated by dynamic programming. J. Dairy Sci. 94: 4476–4487. 14. Schutz, M. M. 1994. Genetic evaluation of somatic cell scores for United States dairy cattle. J. Dairy Sci. 77: 2113–2129. 6. Goto, K., Horiuchi, H., Shinohara, H., Motegi, K., Hashimoto, K., Hongo, S., Gemma, N., Hayashimoto, N., Itoh, T. and Takakura, A. 2007. Specific and quantitative detection of PCR products from Clostridium piliforme, Helicobacter bilis, H. hepaticus, and mouse hepatitis virus infected mouse samples using a newly developed electrochemical DNA chip. J. Microbiol. Methods 69: 93–99. 11. Mori, Y., Nagamine, K., Tomita, N. and Notomi, T. 2001. Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem. Biophys. Res. Commun. 289: 150–154. 1. Bartlett, P. C., Miller, G. Y., Anderson, C. R. and Kirk, J. H. 1990. Milk production and somatic cell count in Michigan dairy herds. J. Dairy Sci. 73: 2794–2800. 10. Losinger, W. C. 2005. Economic impacts of reduced milk production associated with an increase in bulk-tank somatic cell count on US dairies. J. Am. Vet. Med. Assoc. 226: 1652–1658. 13. Okada, J., Horiuchi, H., Hashimoto, K., Hirosawa, D., Kurosaki, Y., Kawamoto, K., Yasuda, J., Makino, S., Gemma, N. and Nikaido, M. 2012. Mobile automatic detection system for bacillus anthracis using electrochemical DNA chip. J. Biosens. Bioelectron. 3: 126. 5. Cremonesi, P., Castiglioni, B., Malferrari, G., Biunno, I., Vimercati, C., Moroni, P., Morandi, S. and Luzzana, M. 2006. Technical note: Improved method for rapid DNA extraction of mastitis pathogens directly from milk. J. Dairy Sci. 89: 163–169. 12. Morin, D. E., Petersen, G. C., Whitmore, H. L., Hungerford, L. L. and Hinton, R. A. 1993. Economic analysis of a mastitis monitoring and control program in four dairy herds. J. Am. Vet. Med. Assoc. 202: 540–548. 7. Hogan, J. S., Gonzalez, R. N., Harmon, R. J., Nickerson, S. C., Oliver, S. P., Pankey, J. W. and Smith, K. L. 1999. Laboratory Handbook on Bovine Mastitis, Revised ed. National Mastitis Council, Madison. 3. Bramley, A. J., Cullor, J. S., Erskine, R. J., Fox, L. K., Harmon, R. J., Hogan, J. S., Nickerson, S. C., Oliver, S. P., Smith, K. L. and Sordillo, L. M. 1996. Microorganisms that cause bovine mastitis. pp. 11–14. In: Current Concepts of Bovine Mastitis, 4th ed. National Mastitis Council, Madison. 2. Beck, H. S., Wise, W. S. and Dodd, F. H. 1992. Cost benefit analysis of bovine mastitis in the UK. J. Dairy Res. 59: 449–460. 9. Kurosaki, Y., Horiuchi, H., Fujinami, Y., Hashimoto, K. and Yasuda, J. 2010. Development and application of rapid and simple method for extracting nucleic acids from microbes. Jpn. J. Forensic Sci. Tech. 15: 135–142. 15. Volling, O. and Krömker, V. 2008. [Udder health management practices in dairy enterprises to reduce the incidence of bovine mastitis]. Dtsch. Tierarztl. Wochenschr. 115: 410–420. 8. Kerro Dego, O., van Dijk, J. E. and Nederbragt, H. 2002. Factors involved in the early pathogenesis of bovine Staphylococcus aureus mastitis with emphasis on bacterial adhesion and invasion. A review. Vet. Q. 24: 181–198. 11 12 13 14 15 1 2 3 4 5 6 7 8 9 10 8449797 - J Am Vet Med Assoc. 1993 Feb 15;202(4):540-8 19024548 - Dtsch Tierarztl Wochenschr. 2008 Nov;115(11):410-20 15906563 - J Am Vet Med Assoc. 2005 May 15;226(10):1652-8 17267057 - J Microbiol Methods. 2007 Apr;69(1):93-9 21854920 - J Dairy Sci. 2011 Sep;94(9):4476-87 2283411 - J Dairy Sci. 1990 Oct;73(10):2794-800 12540135 - Vet Q. 2002 Dec;24(4):181-98 16357279 - J Dairy Sci. 2006 Jan;89(1):163-9 11708792 - Biochem Biophys Res Commun. 2001 Nov 23;289(1):150-4 7929969 - J Dairy Sci. 1994 Jul;77(7):2113-29 1452830 - J Dairy Res. 1992 Nov;59(4):449-60
References_xml	– reference: 3. Bramley, A. J., Cullor, J. S., Erskine, R. J., Fox, L. K., Harmon, R. J., Hogan, J. S., Nickerson, S. C., Oliver, S. P., Smith, K. L. and Sordillo, L. M. 1996. Microorganisms that cause bovine mastitis. pp. 11–14. In: Current Concepts of Bovine Mastitis, 4th ed. National Mastitis Council, Madison. – reference: 10. Losinger, W. C. 2005. Economic impacts of reduced milk production associated with an increase in bulk-tank somatic cell count on US dairies. J. Am. Vet. Med. Assoc. 226: 1652–1658. – reference: 5. Cremonesi, P., Castiglioni, B., Malferrari, G., Biunno, I., Vimercati, C., Moroni, P., Morandi, S. and Luzzana, M. 2006. Technical note: Improved method for rapid DNA extraction of mastitis pathogens directly from milk. J. Dairy Sci. 89: 163–169. – reference: 9. Kurosaki, Y., Horiuchi, H., Fujinami, Y., Hashimoto, K. and Yasuda, J. 2010. Development and application of rapid and simple method for extracting nucleic acids from microbes. Jpn. J. Forensic Sci. Tech. 15: 135–142. – reference: 13. Okada, J., Horiuchi, H., Hashimoto, K., Hirosawa, D., Kurosaki, Y., Kawamoto, K., Yasuda, J., Makino, S., Gemma, N. and Nikaido, M. 2012. Mobile automatic detection system for bacillus anthracis using electrochemical DNA chip. J. Biosens. Bioelectron. 3: 126. – reference: 4. Cha, E., Bar, D., Hertl, J. A., Tauer, L. W., Bennett, G., González, R. N., Schukken, Y. H., Welcome, F. L. and Gröhn, Y. T. 2011. The cost and management of different types of clinical mastitis in dairy cows estimated by dynamic programming. J. Dairy Sci. 94: 4476–4487. – reference: 12. Morin, D. E., Petersen, G. C., Whitmore, H. L., Hungerford, L. L. and Hinton, R. A. 1993. Economic analysis of a mastitis monitoring and control program in four dairy herds. J. Am. Vet. Med. Assoc. 202: 540–548. – reference: 6. Goto, K., Horiuchi, H., Shinohara, H., Motegi, K., Hashimoto, K., Hongo, S., Gemma, N., Hayashimoto, N., Itoh, T. and Takakura, A. 2007. Specific and quantitative detection of PCR products from Clostridium piliforme, Helicobacter bilis, H. hepaticus, and mouse hepatitis virus infected mouse samples using a newly developed electrochemical DNA chip. J. Microbiol. Methods 69: 93–99. – reference: 14. Schutz, M. M. 1994. Genetic evaluation of somatic cell scores for United States dairy cattle. J. Dairy Sci. 77: 2113–2129. – reference: 11. Mori, Y., Nagamine, K., Tomita, N. and Notomi, T. 2001. Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem. Biophys. Res. Commun. 289: 150–154. – reference: 2. Beck, H. S., Wise, W. S. and Dodd, F. H. 1992. Cost benefit analysis of bovine mastitis in the UK. J. Dairy Res. 59: 449–460. – reference: 8. Kerro Dego, O., van Dijk, J. E. and Nederbragt, H. 2002. Factors involved in the early pathogenesis of bovine Staphylococcus aureus mastitis with emphasis on bacterial adhesion and invasion. A review. Vet. Q. 24: 181–198. – reference: 15. Volling, O. and Krömker, V. 2008. [Udder health management practices in dairy enterprises to reduce the incidence of bovine mastitis]. Dtsch. Tierarztl. Wochenschr. 115: 410–420. – reference: 1. Bartlett, P. C., Miller, G. Y., Anderson, C. R. and Kirk, J. H. 1990. Milk production and somatic cell count in Michigan dairy herds. J. Dairy Sci. 73: 2794–2800. – reference: 7. Hogan, J. S., Gonzalez, R. N., Harmon, R. J., Nickerson, S. C., Oliver, S. P., Pankey, J. W. and Smith, K. L. 1999. Laboratory Handbook on Bovine Mastitis, Revised ed. National Mastitis Council, Madison. – ident: 10 doi: 10.2460/javma.2005.226.1652 – ident: 3 – ident: 12 doi: 10.2460/javma.1993.202.04.540 – ident: 14 doi: 10.3168/jds.S0022-0302(94)77154-X – ident: 15 – ident: 5 doi: 10.3168/jds.S0022-0302(06)72080-X – ident: 2 doi: 10.1017/S0022029900027114 – ident: 6 doi: 10.1016/j.mimet.2006.12.009 – ident: 9 doi: 10.3408/jafst.15.135 – ident: 1 doi: 10.3168/jds.S0022-0302(90)78966-7 – ident: 4 doi: 10.3168/jds.2010-4123 – ident: 7 – ident: 13 doi: 10.4172/2155-6210.1000126 – ident: 11 doi: 10.1006/bbrc.2001.5921 – ident: 8 doi: 10.1080/01652176.2002.9695135 – reference: 7929969 - J Dairy Sci. 1994 Jul;77(7):2113-29 – reference: 21854920 - J Dairy Sci. 2011 Sep;94(9):4476-87 – reference: 19024548 - Dtsch Tierarztl Wochenschr. 2008 Nov;115(11):410-20 – reference: 12540135 - Vet Q. 2002 Dec;24(4):181-98 – reference: 17267057 - J Microbiol Methods. 2007 Apr;69(1):93-9 – reference: 16357279 - J Dairy Sci. 2006 Jan;89(1):163-9 – reference: 11708792 - Biochem Biophys Res Commun. 2001 Nov 23;289(1):150-4 – reference: 1452830 - J Dairy Res. 1992 Nov;59(4):449-60 – reference: 8449797 - J Am Vet Med Assoc. 1993 Feb 15;202(4):540-8 – reference: 15906563 - J Am Vet Med Assoc. 2005 May 15;226(10):1652-8 – reference: 2283411 - J Dairy Sci. 1990 Oct;73(10):2794-800
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SubjectTerms	Animals Bacteriology Cattle cow milk DNA chip DNA extraction DNA, Bacterial - isolation & purification Female loop-mediated isothermal amplification (LAMP) mastitis Mastitis, Bovine - diagnosis Mastitis, Bovine - microbiology Milk - chemistry Milk - microbiology Nucleic Acid Amplification Techniques - veterinary pathogen Staphylococcal Infections - diagnosis Staphylococcal Infections - microbiology Staphylococcal Infections - veterinary Staphylococcus aureus Staphylococcus aureus - genetics
Title	Improved rapid and efficient method for Staphylococcus aureus DNA extraction from milk for identification of mastitis pathogens
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