摘要
以16S rRNA基因为检测靶基因,设计10种常见细菌的DNA探针,将探针固定于硝酸纤维素膜条;PCR扩增细菌的16S rRNA基因片段并标记生物素后与膜条杂交;采用碱性磷酸酶标记的链亲和素检测生物素标记,以NBT/BCIP显色。该膜条不仅能单独检测10种细菌中的任何一种,也能同时检测5种细菌。该方法具备高通量、低成本、快速、准确等特点,具有良好的临床应用前景。
16S rRNA gene was taken as test target to design DNA probes for detecting 10 common bacteria then fixed the probes on nitrocellulose membrane strip.16S rRNA gene fragments of the 10 kinds of bacteria were amplified by PCR and labeled with biotin,then hybridized with the membrane strip.The biotin label was checked by SA-AP binding and developed with NBT/BCIP.This membrane strip could detect independently not only anyone of the 10 kinds of bacteria,but also 5 kinds of bacteria at the same time.This method possessed the characters of high-flux,low cost,rapidity and accuracy etc.and had broad future of clinical application.
出处
《微生物学杂志》
CAS
CSCD
2011年第4期58-63,共6页
Journal of Microbiology
基金
湖南省科技计划(2009SK3192)
教育部新教师基金(20090162120028)
中南大学贵重仪器设备开放共享基金(2011-29)
