Evaluation of pulsed-field gel electrophoresis analysis performed at selected prefectural institutes of public health for use in PulseNet Japan.
Evaluation of pulsed-field gel electrophoresis analysis performed at selected prefectural institutes of public health for use in PulseNet Japan.
复制标题
对在选定的县公共卫生机构进行的用于 PulseNet Japan 的脉冲场凝胶电泳分析的评估。
DOI：
10.7883/yoken.jjid.2005.180
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发表时间：
2005
影响因子：
2.2
通讯作者：
Y. Miyazaki
中科院分区：
文献类型：
--
作者：
M. Matsumoto;Yasumoto Suzuki;H. Nagano;J. Yatsuyanagi;Hajime Kurosawa;Kazuhiro Kobayashi;K. Yamaoka;K. Horikawa;J. Kudaka;J. Terajima;Haruo Watanabe;Y. Miyazaki
为了评估在不同县公共卫生机构 (PHI) 中使用脉冲场凝胶电泳 (PFGE) 分析用于 PulseNet Japan 监测系统检测肠出血性大肠杆菌 O157 的可靠性，我们比较了在 8 个选定的 PHI 中对 14 种选定的 O157 菌株进行 PFGE 分型的结果，以评估这些 PHI 中使用的不同实验方案的可靠性。通过使用其自己的方案和/或国家传染病研究所 (NIID) 的方案，对 14 个菌株进行了 PFGE，其中 3 个 PHI 中有 14 个 PFGE 类型，5 个 PHI 中有 13 个 PFGE 类型。 8 个 PHI 中的 5 个的 PFGE 指纹已成功对所有 14 个菌株进行基因分型。当使用 NIID 脉冲方案时，来自一个 PHI 的 PFGE 指纹成功进行了基因分型，但当使用 PHI 自己的方案时则没有进行基因分型。 2 个 PHI 的 PFGE 指纹未能对每个菌株进行基因分型。这种基因分型失败的原因被认为是不适当的 PFGE 脉冲方案或染色体 DNA 消化不充分。这些结果表明 PFGE 协议应该标准化以建立 PulseNet Japan。
In order to evaluate reliability of pulsed-field gel electrophoresis (PFGE) analysis performed at different prefectural public health institutes (PHIs) for use in the PulseNet Japan surveillance system to detect enterohemorrhagic Escherichia coli O157, we compared the results of PFGE-typing of 14 selected strains of O157 performed at 8 selected PHIs to evaluate the reliability of different experimental protocols used in these PHIs. PFGE was performed for 14 strains for which there were 14 PFGE types in 3 PHIs, and 13 PFGE types in 5 PHIs by using their own protocols and/or those of the National Institute of Infectious Diseases (NIID). PFGE fingerprints from 5 out of the 8 PHIs were successfully genotyped for all of the 14 strains. A PFGE fingerprint from one PHI was successfully genotyped when the NIID pulsing protocol was used, but was not genotyped when the PHI's own protocols were used. PFGE fingerprints from 2 PHIs failed to be genotyped for one each of the strains. The cause of this genotyping failure was considered to be inappropriate PFGE pulsing protocols or inadequate digestion of chromosomal DNA. These results suggest that PFGE protocols should be standardized for the establishment of PulseNet Japan.