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

Autor:	UNNO, Hirotaka, INADA, Mika, NAKAMURA, Akiyoshi, HASHIMOTO, Michie, ITO, Keiko, HASHIMOTO, Koji, NIKAIDO, Masaru, HAYASHI, Tomohito, HATA, Eiji, KATSUDA, Ken, KIKU, Yoshio, TAGAWA, Yuichi, KAWAI, Kazuhiro
Jazyk:	angličtina
Rok vydání:	2015
Předmět:	DNA Bacterial Staphylococcus aureus cow milk Bacteriology Staphylococcal Infections Note mastitis Milk Animals Cattle Female loop-mediated isothermal amplification (LAMP) DNA chip DNA extraction Mastitis Bovine Nucleic Acid Amplification Techniques pathogen
Zdroj:	The Journal of Veterinary Medical Science
ISSN:	1347-7439 0916-7250
Popis:	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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=pmid________::5ddb877e26883e556e4acce94b3ed8f4 http://europepmc.org/articles/PMC4565803 Zobrazit plný text záznamu