Quantitative specific detection of Staphylococcus aureus based on recombinant lysostaphin and ATP bioluminescence
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National Natural Science Foundation of China (No. 81072350), China Mega-Project on Major Drug Development (No. 2011ZX09401-023), China Mega-Project on Infectious Disease Prevention (No. 2011ZX10004-001), State Key Laboratory of Pathogen and Biosecurity Program (No. SKLPBS1113).

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    Abstract:

    Quantitative specific detection of Staphylococcus aureus is based on recombinant lysostaphin and ATP bioluminescence. To produce recombinant lysostaphin, the lysostaphin gene was chemically synthesized and inserted it into prokaryotic expression vector pQE30, and the resulting expression plasmid pQE30-Lys was transformed into E. coli M15 for expressing lysostaphin with IPTG induction. The recombinant protein was purified by Ni2+-NTA affinity chromatography. Staphylococcus aureus was detected by the recombinant lysostaphin with ATP bioluminescence, and plate count method. The results of the two methods were compared. The recombinant lysostaphin was successfully expressed, and a method of quantitative specific detection of S. aureus has been established, which showed a significant linear correlation with the colony counting. The detection method developed has good perspective to quantify S. aureus.

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李玉元,米志强,安小平,周育森,童贻刚. 基于重组溶葡球菌酶和ATP生物发光技术特异定量检测金黄色葡萄球菌[J]. Chinese Journal of Biotechnology, 2014, 30(8): 1283-1290

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  • Received:November 12,2013
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  • Online: July 22,2014
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