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PFGE란? #

PFGE란 Pulsed-Field Gel Electrophoresis(펄스필드 젤 전기영동)의 약칭으로 식중독이나 감염증을 일으키는 병원체 간의 감염원 규명에 활용하는 표준 방법으로 병원체의 동정을 위한 DNA fingerprint 기술이다.

PFGE의 원리 #

PFGE uses molecular scissors, called restriction enzymes, to cut bacterial DNA at certain locations known as restriction sites. These molecular scissors are selected to generate a small number of DNA pieces that can be separated based on size. Usually these DNA pieces, or restriction fragments, are large and need to be specially treated and separated to generate a DNA fingerprint. First the bacteria are loaded into an agarose suspension, similar to gelatin, then the bacterial cell is opened to release the DNA. Once the DNA is released then the agarose and DNA suspension, also known as a plug, is treated with restriction enzymes. The treated plugs are then loaded onto an agarose gel and the restriction fragments are separated based on size using an electric field. What makes PFGE different from how scientists usually separate DNA is because PFGE can separate several large restriction fragments. To do this an electric field that constantly changes direction to the gel is used to generate a DNA fingerprint.


PFGE 의의 #

PFGE subtyping has been successfully applied to the subtyping of many pathogenic bacteria and has high concordance with epidemiological relatedness. PFGE has been repeatedly shown to be more discriminating than methods such as ribotyping or multi-locus sequence typingExternal Web Site Icon for many bacteria. PFGE in the same basic format can be applied as a universal generic method for subtyping of bacteria. Only the choice of the restriction enzyme and conditions for electrophoresis need to be optimized for each species. DNA restriction patterns generated by PFGE are stable and reproducible.

PFGE 한계점 #

Time consuming Requires a trained and skilled technician Does not discriminate between all unrelated isolates Pattern results vary slightly between technicians Can’t optimize separation in every part of the gel at the same time Don’t really know if bands of same size are same pieces of DNA Bands are not independent Change in one restriction site can mean more than one band change “Relatedness” should be used as a guide, not true phylogenetic measure Some strains cannot be typed by PFGE

출처 #