【摘要】 无乳链球菌(Streptococcus agalactiae)引起的奶牛乳房炎主要表现为隐性型,需要通过实验室检查才能确诊。研究建立了一种特异、敏感和快速的巢式PCR方法用于检测奶样中的无乳链球菌。利用链球菌属(Streptococcus)16S rRNA基因的保守区设计通用引物作为链球菌属的阳性控制,在保守区内的可变区设计无乳链球菌特异的引物。研究结果表明,通过奶样中无乳链球菌的增菌培养,简便、快速提取DNA和特异性PCR反应,可以检测到奶样中1CFU/mL的无乳链球菌,可以用作牛群中无乳链球菌感染的早期流行病学调查。更多还原

【Abstract】 The bovine mastitis caused by Streptococcus agalactiae is mainly subclinical and can be diagnosed only in the laboratory. A nested PCR-based method for specific , sensitive and rapid detection of Streptococcus agalactiae in raw milk was developed. The general streptococci primers, which anneal to conserved areas within the 16S rRNA subunit gene were used as positive controls. The specificity S. agalactiae primers were designed based on various areas within conserved areas of the 16S rRNA genes of S. agalactiae. The results indicatied that this method enabled the detection of one bacterium in 1 mL of raw milk, after enrichment of Streptococcus agalactiae, DNA extraction using a rapid and simple procedure developed for this purpose, and specific PCR reaction. The developed method could be used for the early infectious investigation of S.agalactiae in the dairy herd and be effectively used in preventing and controlling of S. agalactiae spread in a herd.更多还原