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KMID : 0604219950020010069
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Korean Journal Investigative Dermatology
1995 Volume.2 No. 1 p.69 ~ p.74
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Identification of Vibrio Vulnificus Using Polymerase Chain Reaction
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박석돈
김종구/박재훈/전봉길/최병민/정헌택/김신무.
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Abstract
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Beoause Vibrio vulnificus infection can progress rapidly to threaten life of infected person, rapid identification of this bacterium is essential to reduce the mortality. Recent developments in the polymerase chain reaction (PCR) technique
provides
an
opportunity for rapid and accurate detection of several kinds of bacteria in clinical specimens for the diagnosis of bacterial infections. By using the PCR technique, we screened 76 V. vulnificus strains isolated from septic patients, 8 other
Vibrio
species and 10 non-Vibrio bacterial strains that ca be often cultured from the blood or skin. For DNA extrqaction. Two DNA preparation methods alkali and Kato-Ueno, were compared. We did PCR weth two synthetic primers flanking 340-bp and 519-bp
fragments of cytotoxin-hemolysin gene for amplifying V. vulnificus DNA. Kato-Ueno method was a rapid and efficient method for isolating DNA from V. vunificus compare with alkali method. The PCR products were detected in all V. vulnificus strains
hut
neither in other Vibrio species nor other bacteria. PCR products based on 519-bp DNA frgments were more clearly detected than 340-bp products on agarose gel electrophoresis. These dats suggest that amplification of cytotoxin-hemolysin gene by PCR
is a
highly specific and rapid identification method for the detection of V. vulnificus directly from clinical specimens such as blood or skin lesion without the need for prior isolation of the bacterium.
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KEYWORD
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