[Objective] This study aimed to establish a real-time PCR method for de- tecting stx2 gene in Shiga toxin-producing E. coli （STEC）. [Method] According to the known STEC stx2 gene sequences published in GenBank, PCR ...[Objective] This study aimed to establish a real-time PCR method for de- tecting stx2 gene in Shiga toxin-producing E. coli （STEC）. [Method] According to the known STEC stx2 gene sequences published in GenBank, PCR primers and probes were designed based on the conserved region to construct recombinant plasmid as a positive template, thus optimizing the reaction conditions and establishing the real- time PCR method. [Result] A standard curve was established based on the opti- mized real-time PCR system, indicting a good linear correlation between the initial template concentration and Ct value, with the correlation coefficient F^e of above 0.995. The established method had a good specificity, without non-specific amplifica- tion for 10 non-STEC intestinal bacterial strains; the detection limit of initial template was 1.0x102 copies/μI, indicating a high sensitivity; furthermore, the coefficients of variation within and among batches were lower than 1% and 5% respectively, sug- gesting a good repeatability. [Conclusion] In this study, a real-time PCR method was successfully established for detecting STEC stx2 gene, which provided technical means for rapid detection of STEC in samples.展开更多
China Oceans Law Review (COLR – ISSN:1813-7350) is a compilation of research focusing on ocean-related laws and policies. This Chinese-English bilingual journal is jointly hosted by Xiamen University, Hong Kong Polyt...China Oceans Law Review (COLR – ISSN:1813-7350) is a compilation of research focusing on ocean-related laws and policies. This Chinese-English bilingual journal is jointly hosted by Xiamen University, Hong Kong Polytechnic University, University of Macao Institute for Advanced Legal Studies, (Taiwan) Sun Yat-sen University Institute of Marine Affairs and Dalian Maritime University Institute of Maritime Law and Ocean Law, and is published semi-annually. Beginning from Spring 2011,COLR is indexed by Westlaw China, Lawinfochina. com, Airiti, Cqvip and Heinonline.展开更多
基金Supported by Agricultural Science and Technology Support Program(Social Development)of Jiangsu Province(BE2011771)~~
文摘[Objective] This study aimed to establish a real-time PCR method for de- tecting stx2 gene in Shiga toxin-producing E. coli （STEC）. [Method] According to the known STEC stx2 gene sequences published in GenBank, PCR primers and probes were designed based on the conserved region to construct recombinant plasmid as a positive template, thus optimizing the reaction conditions and establishing the real- time PCR method. [Result] A standard curve was established based on the opti- mized real-time PCR system, indicting a good linear correlation between the initial template concentration and Ct value, with the correlation coefficient F^e of above 0.995. The established method had a good specificity, without non-specific amplifica- tion for 10 non-STEC intestinal bacterial strains; the detection limit of initial template was 1.0x102 copies/μI, indicating a high sensitivity; furthermore, the coefficients of variation within and among batches were lower than 1% and 5% respectively, sug- gesting a good repeatability. [Conclusion] In this study, a real-time PCR method was successfully established for detecting STEC stx2 gene, which provided technical means for rapid detection of STEC in samples.
文摘China Oceans Law Review (COLR – ISSN:1813-7350) is a compilation of research focusing on ocean-related laws and policies. This Chinese-English bilingual journal is jointly hosted by Xiamen University, Hong Kong Polytechnic University, University of Macao Institute for Advanced Legal Studies, (Taiwan) Sun Yat-sen University Institute of Marine Affairs and Dalian Maritime University Institute of Maritime Law and Ocean Law, and is published semi-annually. Beginning from Spring 2011,COLR is indexed by Westlaw China, Lawinfochina. com, Airiti, Cqvip and Heinonline.
