최확수법과 중합효소연쇄반응을 이용한 비브리오 패혈증균 정량분석법 확립 및 검증

Abstract

Vibrio vulnificus is an opportunistic pathogen which is highly lethal and responsible for the majority of seafood -associated deaths worldwide. The common method used for the detection of V. vulnificus was utilized thiosulfate citrate bile salts sucrose (TCBS). The objective of this study is to establish a quantitative count method of V. vulnificus, and the most frequent method for quantitative cell count was plate count method. However, using TCBS agar plate for viable cell count of V. vulnificus is not suitable to detect the damaged V. vulnificus due to growth inhibition by high selectivity. In this study, I suggested a most probable number-polymerase chain reaction (MPN-PCR) method using alkaline peptone water medium for quantitative cell count of V. vulnificus. And the MPN-PCR method developed in this study was used to analyze the effect of salinity, temperature and organic acid-adjusted pH on V. vulnificus. The MPN-PCR method showed 2 log higher cell number than that of TCBS agar plate. Similar result was also found in the control, which is Luria-Bertani agar containing 2% NaCl. As a result of analyzing the environmental factors using the MPN-PCR method, V. vulnificus was more salinity-dependent than temperature. It can also be seen that low pH such as pH 4, 5 affects V. vulnificus growth. Among them, the water adjusted with acetic acid was controlled V. vulnificus the greatest. Therefore, the MPN-PCR method can be utilized a sensitive method for the quantitative detection of viable cell count of V. vulnificus in fish as well as shellfish samples. In addition, it is possible to suggest a method for enhancing the hygienic safety of aquatic products by conducting V vulnificus control research.