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標題: 食物中毒相關之金黃色葡萄球菌分析
Food poisoning related to analysis of Staphylococcus sureus
作者: 林穎君
Lin, Ying-Chung
Contributors: 陳錦樹
食品暨應用生物科技學系所
關鍵字: 金黃色葡萄球菌;生化鑑定;複合式聚合酶鏈鎖反應;脈衝式電泳
Staphyococcus aureus;Biochemical identification;Multiplex PCR;pulsed-field gel electrophoresis
日期: 2013
Issue Date: 2013-11-18 14:22:10 (UTC+8)
Publisher: 食品暨應用生物科技學系所
摘要: 食物中毒案件中最常見的為細菌性食物中毒,而其中金黃色葡萄球菌 (Staphylococcus aureus) 是很嚴重的細菌性食物中毒菌。近年金黃色葡萄球菌對 methicillin 有抗藥性之比例急速增加,而對methicillin具有抗藥性的金黃色葡萄球菌即稱為MRSA (Methicillin-resistant Staphylococcus aureus )。MRSA 是全球臨床上極受重視的多重抗藥菌。MRSA的產生是金黃色葡萄球菌獲得一移動性的基因片段: staphylococcal cassette chromosome mec (SCCmec) 對 methicillin 產生抗性。本研究收集98、99年通報食物中毒個案檢體3290件,經臨床培養、分離,以生化鑑定︰staphylase test、凝固酶試驗、觸酶試驗、API ID 32 STAPH 生化鑑定、VITEK 2 革蘭氏陽性菌鑑定組鑑定確認為金黃色葡萄球菌,再以Enterotoxin-F菌株毒素鑑定套組件測定其腸毒素型,117株金黃色葡萄球菌中腸毒素型別中確認出A型 49%、B型 32%、C型 18%、D型 1% 毒素型菌株。而由複合式聚合酶鏈鎖反應中得知含Sccmec基因者有 21%,再進一步分析,發現第三型佔 8%,第四型76%,未分出型別者 16%,由結果顯示76% 為社區感染型。接著以脈衝式電泳分析,分別以地區、腸毒素型、含SCCmec 基因、醫院別進行比對,結果顯示:大多無區別性,可能是因為是由非特定條件中所篩選出之菌株所致。最後進行藥物敏感性試驗,藥敏結果顯示菌株之抗藥性MRSA菌株對於Amoxicillin/Clavulanate、Ampicillin、Cefazolin、Cefoxitin、Oxacillin、Penicillin G已有100%耐藥性,而Daptomycin、Linezolid、Mupirocin High level、Nitrofurantoin、Quinupristin-dalfopristin、Rifampin、Teicoplanin、Trimethoprim-Sulfamethoxazole、Vancomycin等有100%敏感,此次加入之兩種新測試藥物發現:MRSA已經對Cefazolin 產生了抗藥性,而Daptomycin則是現在還可以使用在治療上的藥物。
The most common food poisoning cases of bacterial food poisoning, including staphylococcus aureus is a very serious bacterial food poisoning bacteria. In recent years, the proportion of drug resistance of methicillin in staphylococcus aureus in crease rapidly,and we called methicillin resistant staphylococcus aureus (MRSA). MRSA is very important multi-drug-resistant bacteria in global clinical. MRSA generated is staphylococcus aureus obtained a mobility of a gene fragment: staphylococcal cassette chromosome mec (SCCmec). We collected 3290 food poison cases in 2009 and 2010 . By clinical culture ,separation, biochemical identification ︰ staphylase test, coagulase test, catalase test, API ID 32 STAPH biochemical identification, VITEK 2 Gram-positive bacteria identified kit were recognized as staphylococcus aureus, and enterotoxin type is determined to the Enterotoxin-F strains toxins identification kits.There are identified 117 staphylococcus aureus enterotoxin type : type A 49 %, type B 32 %, type C 18 %, type D 1 % toxin type strain. 21 % was pound containing SCCmec gene with the Multiplex -polymerase chain reaction. And further analysis, the third type was 8%, the fourth type was 76 %, and not-type was 16 %. The results show that 76 % was community-acquired type. Pulsed-field gel electrophoresis(PFGE) was performed to assess and compare by the region , enterotoxin type, containing SCCmec gene and hospitals respectively. The data showed : mostly without distinction. Because of the strains may be screened out with non-specific conditions. Finally, the results of drug sensitivity test susceptibility showed MRSA strains had 100% resistance of Amoxicillin / Clavulanate, Ampicillin, Cefazolin, Cefoxitin, Oxacillin and Penicillin G, and that had 100% sensitive of Daptomycin linezolid, Mupirocin High level, Nitrofurantoin, Quinupristin-dalfopristin, Rifampin, Teicoplanin, Trimethoprim-Sulfamethoxazole and Vancomycin. These results suggested that MRSA had resistance of Cefazolin and Daptomycin can also be used for the treatment of drug.
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