超广谱TEM-内酰胺酶检测及分型

Detection and typing of plasmid mediated TEM extended spectrum β lactamase

目的 建立有效和实用的方法，检测质粒介导的ＴＥＭ 型超广谱β内酰胺酶并对其分型。方法 采用聚合酶链反应序列特异性寡核苷酸探针杂交（ＰＣＲＳＳＯ） 技术，首先用ＴＥＭ β内酰胺酶基因的通用引物进行扩增，再用序列特异性寡核苷酸探针与扩增产物进行杂交，对多株标准产酶菌株进行分型。结果 产ＴＥＭβ内酰胺酶的菌株扩增阳性，而产其他β内酰胺酶的菌株扩增阴性，探针杂交的结果符合标准菌株的产酶型别。结论 ＰＣＲＳＳＯ 是一种有效的检测ＴＥＭ 超广谱β内酰胺酶及分型的方法。为临床检验、流行病学调查和研究提供了依据。

Objective To establish an effective method for the detection and typing of TEM extended spectrum β lactamase(ESBL). Methods The polymerase chain reaction sequence specific oligonucleotide (PCR SSO) was used. PCR was performed with β lactamase common primers. Then the PCR production was hybridized with digoxigenin labeled sequence specific oligonucleotide. Results Strains which produce TEM β lactamase were positive by PCR, whereas others were negative. The oligotypings which determined by hybridization accorded with their real kinds. Conclusion The PCR SSO is an effective method for the detection and typing of TEM ESBL, can be used in clinical detection and epidemiological study .