Chelex-100法直接提取乳样中奶牛乳房炎主要致病菌DNA方法的建立

Abstract

Abstract: In this research,a simple and rapid extraction of microbial DNA for PCR using Chelex-100 chelating resin directly from milk was built.3 common mastitis pathogens (S.aureus,E.coli and S.agalactiae) were submitted to tenfold serial dilu-tions and artificially inoculated to milk samples.Then the sensitivities of the PCR assay using the Chelex-100 DNA extraction method were detected.In the meantime,the sensitivity of the 3 common mastitis using phenol-chloroform DNA extraction method was compared.The results showed that the PCR assay using the Chelex-100 DNA extraction method was more sensitive than that using phenol-chloroform DNA extraction method.The sensitivity of the PCR assay using the Chelex-100 DNA extraction method in detecting S.aureus, E.coli and S.agalactiae was 10³,10² and 10² CFU/mL,repectively.The sensitivity of the PCR assay using phenol-chloroform DNA extraction method in detecting S.aureus,E.coli and S.agalactiae was all 10⁴CFU/mL.The thresholds of the sensitivities using the Chelex-100 DNA extraction method were compatible with their use as an efficient tool of diagnosis in bovine mastitis.The Chelex-100 DNA extraction method was simple,rapid,cheap,involve no organic solvents and meaningful for direct extraction of DNA of other pathogens from milk.Undoubtedly,this method will make sense for rapid diagnose of bovine mastitis by PCR.

Key words: Chelex-100; bovine mastitis; bacterial DNA; PCR; sensitivity

CLC Number:

S858.23

XU Jin-peng, HAO Yong-qing. Development of Chelex-100 as a Medium for Simple DNA Extraction of Major Mastitis Pathogens Directly from Milk[J]. China Animal Husbandry & Veterinary Medicine, 2014, 41(11): 25-29.

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Development of Chelex-100 as a Medium for Simple DNA Extraction of Major Mastitis Pathogens Directly from Milk

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