刘祥, 张玲, 李新琼, 闫国栋, 邹年莉, 张正东, 李群, 熊衍文, 王红. 艾伯特埃希菌快速鉴定方法比较研究[J]. 疾病监测, 2017, 32(8): 678-682. DOI: 10.3784/j.issn.1003-9961.2017.08.016
引用本文: 刘祥, 张玲, 李新琼, 闫国栋, 邹年莉, 张正东, 李群, 熊衍文, 王红. 艾伯特埃希菌快速鉴定方法比较研究[J]. 疾病监测, 2017, 32(8): 678-682. DOI: 10.3784/j.issn.1003-9961.2017.08.016
LIU Xiang, ZHANG Ling, LI Xin-qiong, YAN Guo-dong, ZOU Nian-li, ZHANG Zheng-dong, LI Qun, XIONG Yan-wen, WANG Hong. Comparative study on rapid identification methods for Escherichia albertii[J]. Disease Surveillance, 2017, 32(8): 678-682. DOI: 10.3784/j.issn.1003-9961.2017.08.016
Citation: LIU Xiang, ZHANG Ling, LI Xin-qiong, YAN Guo-dong, ZOU Nian-li, ZHANG Zheng-dong, LI Qun, XIONG Yan-wen, WANG Hong. Comparative study on rapid identification methods for Escherichia albertii[J]. Disease Surveillance, 2017, 32(8): 678-682. DOI: 10.3784/j.issn.1003-9961.2017.08.016

艾伯特埃希菌快速鉴定方法比较研究

Comparative study on rapid identification methods for Escherichia albertii

  • 摘要: 目的 评价目前用于快速鉴定艾伯特埃希菌方法的特异性。方法 利用51株艾伯特埃希菌及其他对照菌株,分别评价基于clpX、lysP和mdh基因的三重聚合酶链式反应(PCR);基于EA 0134基因的单重PCR;基于EACBF 2103-EACBF2104基因的巢式PCR。结果 51株艾伯特埃希菌三重PCR均为阳性,鉴定吻合率为100%;巢式PCR两轮结果均为阳性,鉴定吻合率为100%,对照组第1轮PCR结果全部为阴性,但第2轮出现非特异条带。51株中49株EA 0134基因为阳性,鉴定吻合率为96.08%,对照菌株中伤寒沙门菌和肠出血性大肠埃希菌O157:H7扩增出大小类似的片段。结论 EA 0134基因在一些菌株中缺失,会造成漏判,同时在部分肠道菌株中存在非特异性扩增,会造成误判。三重PCR和巢式PCR的第1轮扩增均具有高特异性,可作为目前艾伯特埃希菌分离株的PCR快速鉴定方法,为艾伯特埃希菌相关的临床诊断和突发公共卫生事件处置,提供准确快速的诊断依据。

     

    Abstract: Objective To understand the specificity of the rapid identification methods for Escherichia albertii. Methods Fifty-one E. albertii strains and other control strains were used to evaluate the specificities of multiplex-PCR based on clpX, lysP and mdh genes, PCR based on EA 0134 gene and nested PCR based on EACBF 2103 -EACBF 2104 gene. Amplification sequences were used for analysis and compare after sequencing. Results All of 51 strains of E. albertii were positive in multiplex-PCR, the identification rate was 100%.The results of 2 round nested PCR were all positive, the identification rate was 100%, and the first round nested PCR results in the control group were all negative, but non-specific band appeared in the second round. In the 51 strains, 49 were positive for EA 0134 gene, the identification rate was 96.08%, and similar fragments were amplified from Salmonella typhi and Escherichia coli O157:H7 in the control strains. Conclusion The deletion of EA 0134 gene in some strains might lead to misdiagnosis. The multiplex-PCR and 1st round nested PCR were highly specific, which can be used as the main method for the rapid identification of suspected E. albertii strains, clinical diagnosis of E. albertii infection and response of related public health emergency.

     

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