新設計聚合酶連鎖反應引子對金黃色葡萄球菌腸毒素C1、C2及C3型基因之檢測   全文下載

Novel PCR Primers for Specific Detection of C1, C2 and C3 Enterotoxin Genes in Staphylococcus aureus

金黃色葡萄球菌是引起臨床感染及食物中毒的一主要病原菌，其所產生之腸毒素，包括腸毒素A、B、C、D、E、G、H、I、J、K、L和M型等，其中腸毒素C型依抗原性可進一步區分為三個主要的亞型C1、C2、C3及其他亞型。雖然這些腸毒素C1、C2及C3基因亞型的核?酸序列同源性相似程度高達97%，但我們仍可發展出第二套聚合?連鎖反應引子，用以區分腸毒素C1、C2及C3亞型。這套PCR引子之組合分別為C12F／C1BR, C12F／C23R及C3BF／C23R，可用來檢測金黃色葡萄球菌腸毒素C1、C2及C3亞型。利用所設計的引子檢測1995-1997年間於中台灣所發生食物中毒案例中食物及病人檢體所分離的39株腸毒素C型金黃色葡萄球菌菌株，其檢測結果與先前所發表的引子組之檢測結果完全一致，可見目前的此組引子組可用來檢測這三種亞型腸毒素基因。"

Staphylococcus aureus is the major pathogen that causes clinical infection as well as food poisoning. Enterotoxin produced by Staphylococcus aureus strains are, among others, staphylococcal enterotoxin (SE) A, SEB, SEC, SED, SEE, SEG, SEH, SEI, SEJ, SEK, SEL and SEM. For SEC, there are three major antigentically distinct SEC subtypes, i.e. SEC1, SEC2 and SEC3, in addition to other molecular variants. The nucleotide sequence homology between SEC1, SEC2, and SEC3 genes is higher than 97%; however, we were able to develop a second set of PCR primers that allowed us to differentiate the three SECs, i.e., SEC1, SEC2 and SEC3, S. aureus strains. These PCR primers and their combinations were C12F/C1BR, C12F/C23R and C3BF/C23R, for specific detection of the SEC1, SEC2 and SEC3 genes of S. aureus strains, respectively. Using these primers, we examined 39 SEC S. aureus isolates, which were obtained from foods that were likely the source of food-borne outbreaks between 1995 and 1997 in central Taiwan, and from the patients of the outbreaks. The results were consistent with those obtained from the first set of primers reported earlier, indicating that the present primers could be used for specific detection of these three toxin types. 金黃色葡萄球菌是引起臨床感染及食物中毒的一主要病原菌，其所產生之腸毒素，包括腸毒素A、B、C、D、E、G、H、I、J、K、L和M型等，其中腸毒素C型依抗原性可進一步區分為三個主要的亞型C1、C2、C3及其他亞型。雖然這些腸毒素C1、C2及C3基因亞型的核苷酸序列同源性相似程度高達97%，但我們仍可發展出第二套聚合酶連鎖反應引子，用以區分腸毒素C1、C2及C3亞型。這套PCR引子之組合分別為C12F／C1BR, C12F／C23R及C3BF／C23R，可用來檢測金黃色葡萄球菌腸毒素C1、C2及C3亞型。利用所設計的引子檢測1995-1997年間於中台灣所發生食物中毒案例中食物及病人檢體所分離的39株腸毒素C型金黃色葡萄球菌菌株，其檢測結果與先前所發表的引子組之檢測結果完全一致，可見目前的此組引子組可用來檢測這三種亞型腸毒素基因。