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Evaluation of Pulsed-Field Gel Electrophoresis Analysis Performed at Selected Prefectural Institutes of Public Health for Use in PulseNet Japan

2005; National Institute of Infectious Diseases; Volume: 58; Issue: 3 Linguagem: Inglês

10.7883/yoken.jjid.2005.180

1884-2836

Masakado Matsumoto, Yasumoto Suzuki, Hideki Nagano, Jun Yatsuyanagi, Hájíme Kurosawa, Kazuhiro Kobayashi, Koji Yamaoka, Kazumi Horikawa, Jun Kudaka, Jun Terajima, Haruo Watanabe, Yutaka Miyazaki,

Listeria monocytogenes in Food Safety

In order to evaluate the 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.