Use of primers based on the heat shock protein genes hsp70, hsp40, and hsp10, for the detection of bovine mastitis pathogens Streptococcus agalactiae, Streptococcus uberis and Streptococcus bovis.
Streptococcus agalactiae, Streptococcus uberis, and Streptococcus bovis are three of the major pathogens which cause mastitis in dairy herds. Since conventional methods for the detection of these mastitis pathogens are laborious and time-consuming, rapid methods are needed. With an attempt to know if heat shock protein (HSP) genes other than HSP60 gene, could be used for PCR primer designing, in this study, we tried to design PCR primers based on the heat shock protein genes hsp70, hsp40, and hsp10 for the specific detection of S. agalactiae, S. uberis, and S. bovis, respectively. Using these primers, all the randomly selected target strains could be specifically detected. Bacterial species other than the target organisms, including strains of other Streptococcus spp., and strains of non-Streptococcus spp., would not generate any false positive results. As these PCR primers were used for direct detection of mastitis pathogens, the detection limit was N (N=1-9) x 10(3)CFU/ml of cell dilutions. If a 10h pre-enrichment step was performed, the detection limit was N x 10(0)CFU/ml. Thus, these primers could be used for the specific and sensitive detection of bovine mastitis bacteria.
Chiang
Yu-Cheng
YC
Institute of Industrial Biotechnology, Hung Kuang University, Shalu, Taichung County 433, Taiwan, ROC.
Pai
Wan-Yu
WY
Chen
Chih-Yuan
CY
Tsen
Hau-Yang
HY
D000818:Animals; D002417:Cattle; D018835:Chaperonin 10 / Q000032:analysis / Q000235:genetics; D017931:DNA Primers / Q000032:analysis* / Q000235:genetics*; D050956:HSP40 Heat-Shock Proteins / Q000032:analysis / Q000235:genetics; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis / Q000235:genetics; D006360:Heat-Shock Proteins / Q000032:analysis* / Q000235:genetics*; D008414:Mastitis, Bovine / Q000382:microbiology*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013291:Streptococcus / Q000145:classification / Q000235:genetics* / Q000472:pathogenicity*
D018835:Chaperonin 10; D017931:DNA Primers; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins