An improved procedure for the pulsed-field gel electrophoresis analysis of Vibrio vulnificus
Vibrio vulnificus is a marine bacterium that causes septicemia with high mortality via wound infection or seafood ingestion. Subspecies typing of V. vulnificus can be performed by pulsed-field gel electrophoresis (PFGE). However, this method is hampered by the degradation of chromosomal DNA in about 12% of strains. This study presents a modified PFGE procedure. The outer membrane of V. vulnificus cells was lysed utilizing a sucrose-EDTA method and components in the periplasmic space were removed prior to making plugs. Cytoplasmic digestive enzymes were inactivated by an extended proteinase K reaction. Experimental results indicated that five out of eight strains exhibiting smeared DNA in a previous study (Appl. Environ. Microbiol. 70: 5153-5158. 2004) achieved clear banding patterns. This modified PFGE procedure can be applied to improve PFGE typing of untypeable strains of V. Vulnificus.
Wong, H.-C. and Kuo, C.-J.
"An improved procedure for the pulsed-field gel electrophoresis analysis of Vibrio vulnificus,"
Journal of Food and Drug Analysis: Vol. 14
, Article 1.
Available at: https://doi.org/10.38212/2224-6614.2480