Standardization and International Multicenter Validation of a PulseNet Pulsed-Field Gel Electrophoresis Protocol for Subtyping Shigella flexneri Isolates

Details

• Alternative Title:
  Foodborne Pathog Dis
• Personal Author:
  Pichel, Mariana ; Brengi, Silvina P. ; Cooper, Kara L.F. ; Ribot, Efrain M. ; Al-Busaidy, Suleiman ; Araya, Pamela ; Fernández, Jorge ; Vaz, Tania Ibelli ; Kam, Kai Man ; Morcos, Myriam ; Nielsen, Eva M. ; Nadon, Celine ; Pimentel, Guillermo ; Pérez-Gutiérrez, Enrique ; Gerner-Smidt, Peter ; Binsztein, Norma
• Corporate Authors:
  Shigella flexneri PulseNet PFGE Protocol Working Group
• Description:
  Shigella flexneri is one of the agents most frequently linked to diarrheal illness in developing countries and often causes outbreaks in settings with poor hygiene or sanitary conditions. Travel is one of the means by which S. flexneri can be imported into developed countries, where this pathogen is not commonly seen. A robust and discriminatory subtyping method is needed for the surveillance of S. flexneri locally and regionally, and to aid in the detection and investigation of outbreaks. The PulseNet International network utilizes standardized pulsed-field gel electrophoresis (PFGE) protocols to carry out laboratory-based surveillance of foodborne pathogens in combination with epidemiologic data. A multicenter validation was carried out in nine PulseNet laboratories located in North and South America, Europe, and Asia, and it demonstrated that a new protocol is highly robust and reproducible for subtyping of S. flexneri. This protocol, already approved for PulseNet laboratories, applies NotI and XbaI as primary and secondary restriction enzymes, respectively, under electrophoresis conditions of initial switch time of 5 s to final switch time of 35 s, at 6 volts/cm.
• Subjects:
  Bacterial Typing Techniques Denmark Deoxyribonucleases, Type II Site-Specific DNA, Bacterial Dysentery, Bacillary Electrophoresis, Gel, Pulsed-Field Hong Kong Middle East North America Quality Control Reproducibility Of Results Shigella Flexneri South America Time Factors

  More +
• Source:
  Foodborne Pathog Dis. 9(5):418-424
• Pubmed ID:
  22506731
• Pubmed Central ID:
  PMC11359307
• Document Type:
  Journal Article
• Funding:
  CC999999/ImCDC/Intramural CDC HHSUnited States/
• Volume:
  9
• Issue:
  5
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha-512:d3972953ff0489442b0caeff7e94d3232af3d417f6b09447c783eab2f0212d3349af18a6f5fc3f425dde191705e764fc4bf46b0301cce0461afb060217081fd6
• Download URL:
  https://stacks.cdc.gov/view/cdc/160687/cdc_160687_DS1.pdf
• File Type:
  [PDF - 629.79 KB ]