Establishment of Nested PCR of Streptococcus uberis for the Cow Subclinical Mastitis

In order to research sensitivity that detect cow subclinical mastitis by nested PCR, Streptococcus uberis, Streptococcus agalactiae, Escherichia coli and Staphylococcus aureus separated from fresh dairy samples which have been identified by physiological and biochemical methods were used as tested materials. In this experiment, the bacterial DNA was extracted, then universal primer was designed according to bacteria 16S rRNA conserved region, nested PCR technique was used to amplify the nucleotide sequence. The specificity primers were designed based on various areas within conserved areas of the 16S rRNA genes of four pathogenic bacteria. The result showed nested PCR technique could detect various areas of conserved areas of the 16S rRNA genes of S. uberis. The result was the same as physiological and biochemical methods. This theory could be rapidly and efficiently applied in detecting S. uberis for the cow subclinical mastitis.