金黃色葡萄球菌 ( Staphylococcus aureus )是造成食物中毒的重要病原菌之一，其產生的腸毒素 (enterotoxin ) 依照血清學上的分類可分為SEA、SEB、SEC、SED、SEE五種傳統型腸毒素與中毒性休克症候群毒素 ( TSST-1 )，而近年來有許多的新型腸毒素被發現，如：SEG、SEH、SEI、SEJ、SEK、SEL、SEM、SEN、SEO、SEP、SEQ、SER及SEU等。本研究目的在於設計用於PCR之新型腸毒素基因sen、seo和sep之特異性引子組，並對金黃色葡萄球菌臨床分離株進行新型腸毒素基因的檢測，來探討新型腸毒素基因的分佈情形，且同時使用已發表之TSST-1引子組，對金黃色葡萄球菌臨床菌進行分佈調查，同時了解其分佈與傳統型腸毒素基因的關聯性。
本研究中針對新型腸毒素所設計的引子組可以正確篩選出腸毒素基因sen、seo和sep。利用非金黃色葡萄球菌之葡萄球菌屬菌株及其他的腸內菌進行PCR檢測，結果得知除了金黃色葡萄球菌之實驗室標準菌會有PCR產物( sen, 135 bp; seo, 172 bp; sep, 148 bp )之外，其餘的非金黃色葡萄球菌皆無PCR產物的出現。至於在靈敏度的測試方面，在未進行增殖培養的條件下，引子組的PCR檢測靈敏度可達到N╳103~104 CFU/ml，而另外進行10小時的增殖培養後，引子組的PCR靈敏度更增加到約N╳100 CFU/ml，由以上的結果可得知引子組利用PCR，篩檢金黃色葡萄球菌腸毒素基因sen、seo、sep其靈敏度佳。本研究也另外針對食品樣本來了解引子組檢測的靈敏度，我們選用市售鮮乳及肉品以額外添加的方式加入金黃色葡萄球菌，在添加之前菌株先以TSB預培養10小時，然後以引子組檢測鮮乳及肉品中之金黃色葡萄球菌腸毒素基因sen、seo、sep，而其靈敏度也可達到N╳100 CFU/g，結果表示本研究所設計的特異性引子組，可應用在食品樣品的檢測。
本研究對於帶有金黃色葡萄球菌新型腸毒素基因sen、seo及sep的臨床菌，進行反轉錄聚合鏈鎖反應( RT-PCR )，我們各選11株PCR為正反應的金黃色葡萄球菌，實驗結果顯示，原本帶有腸毒素基因的菌株利用RT-PCR檢測皆會有mRNA的表現。
關於調查金黃色葡萄球菌毒素性休克症候群毒素( TSST-1 )和新型腸毒素( SEN、SEO、SEP )基因的分佈相關性，所採用的臨床菌株的來源，為台中榮民總醫院及疾病管制局第三分局所提供的金黃色葡萄球菌282株，而帶有TSST-1毒素基因的菌株總共佔126株 ( 44.6 % )，其中同時帶有sea或sec的菌株共有48株 (38 %)；腸毒素基因sen有40株 ( 14 % )，帶有腸毒素基因seo有44株 ( 15.6 % )，帶有腸毒素基因sep有91株 ( 32 % )，在同時具有sen和seo的29株菌株中，有15株是連同sec一起出現。
我們藉腸毒素基因的分佈情形，可了解腸毒素基因之間的關聯性，例如當檢測出帶有TSST-1毒素基因的菌株，往往也都有腸毒素基因sea或sec的出現，因此TSST-1毒素基因的分佈甚為廣泛，以及本研究所針對的sen,seo和sep腸毒素基因，在臨床及食物中毒分離株的分佈情況中，也發現帶有sep腸毒素基因的菌株數量比帶有sen和seo腸毒素基因的菌株數要多；另外帶有腸毒素基因sen及seo的菌株中，也能發現腸毒素基因seg、sei和sem的存在，這可說明金黃色葡萄球菌通常帶有一種或一種以上的腸毒素基因。

Staphylococcus aureus is one of the important pathogens which cause food poisoning. The staphylococcal enterotoxins (SEs) produced by strains of S. aureus can be divided into SEA, SEB, SEC, SED, SEE, classical enterotoxins, and toxic shock syndrome toxin (TSST-1). Recently, many new type of SEs have been found, for example: SEG, SEH, SEI, SEJ, SEK, SEL, SEM, SEN, SEO, SEP, SEQ, SER and SEU, etc. The purposes of this study is to design specific primers for SEN, SEO and SEP genes, detection and use them for the survey of the distribution of SE genes and TSST of S. aureus isolates in Taiwan.
In this study, PCR primers specific for sen, seo and sep are designed. Non-staphylococcus species and staphylococcus species other than S. aureus would not generate any fales positive results. Moleclar sizes of the PCR products were 135 bp for sen, 172 bp for seo and 148 bp for sep respectively. Detection sensitivity was N╳103~104 CFU/ml, for samples without preincubation, and N╳100 CFU/ml for samples with the preculture step. These primers were also used for the monitoring of sen, seo and sep S.aureus strains in food smples, such as whole milk and pork.
Eleven PCR positive strains of S. aureus were also assayed by RT-PCR. It was found that all these randomly selected S. aureus strains with sen, seo and sep genes were positive by RT-PCR.
To investigate the distribution of TSST-1 and new SEs (SEN , SEO , SEP) genes in S. aureus, 284 isolates obtained from Veternary General Hospital in Taichung and Center for Disease Control, the third branch in Taiwan, were analyzed by PCR. In 126 isolates (44.3 %)found positive for TSST-1 gene, 48 isolates (38%) were found positive for sea and sec. Forty isolates were positive for sen, 44 for seo and 91 for sep. In 29 isolates with sen and seo, 15 strains were with sec.
Results of SE distribution study, indicate that strains with TSST-1 toxin gene are often with sea or sec. The ratio of sep producing S. aureus strains are higher than the sen and seo producing strains. Coexist of seg, sei and sem in strains with sen and seo were found. According to the above facts, S aureus strains are often found with one or more SEs genes.

目 次
頁次
中文摘要………………………………………………………………….…………I
英文摘要……………………………………………………………………………III
Ι、文獻整理………………………………………………………………………1
一、葡萄球菌的特色及所造成的疾病…………………………………….....1
二、金黃色葡萄球菌之檢測……………………………………………….....3
三、金黃色葡萄球菌產生的毒性因子…………………………………….. ...4
四、葡萄球菌的抗生素耐性………………………….………..………………7
五、金黃色葡萄球菌腸毒素分佈情形…………………………………….....7
六、各型腸毒素之介紹…………………………………..................................8
1. A型腸毒素 （SEA）……………………………………………........8
2. B型腸毒素 （SEB）……………………………………………........9
3. C 型腸毒素 （SEC）……………………………………………........9
4. D 型腸毒素 （SED）……………………………………………......10
5. E 型腸毒素 （SEE）……………………………………………….11
6. G 型腸毒素 （SEG）……………………………………………......11
7. H 型腸毒素 （SEH）…………………………………………...........11
8. I 型腸毒素 （SEI）…………………………………..………..........12
9. J型腸毒素 （SEJ）…………………………………………............12
10. K型腸毒素 （SEK）…………………………………………….......12
11. L型腸毒素 （SEL）………………………………………..………..13
12. M型腸毒素 （SEM）………………………………………………..13
13. N,O型腸毒素 (SEN, SEO)……………………….…………………..14
14. P型腸毒素 (SEP)……………………………………….…………….14
15. Q型腸毒素 （SEQ）……………………………………………........15
16. R型腸毒素 (SER)……………………………………………………..15
17. U型腸毒素 (SEU)……………………………………………….........16
七、金黃色葡萄球菌腸毒素之檢測………………………...............................16
Ⅱ、金黃色葡萄球菌新型腸毒素N、O、P基因引子組之設計及其應用於食品及臨床菌株腸毒素型之分佈調查和應用
壹、前言…………………………………………………………………………22
貳、材料與方法……………………………………………….………………..24
一、 實驗材料……………………………………………………………...24
(一) 菌種………………………………………………………………….24
(二) 培養基……………………………………………………………….24
(三) 藥品………………………………………………………………….25
(四) 緩衝液及試劑……………………………………………………….26
(五) 儀器………………………………………………………………….27
二、 實驗方法……………………………………………………………...27
(一) PCR引子組之設計與合成…………………………………………..27
(二) 聚合鏈鎖反應 ( PCR )……………………………………………27
1、 葡萄球菌的保存與鑑定……………………………………....28
2、 葡萄球菌DNA之製備………………………………………..28
(1) Phenol/ chloroform 法…………………………………………28
(2) 商業套組法 ( Viogene, Blood & Tissue Genomic Mini )……..28
3、 非葡萄球菌DNA之製備 ( Viogene, Blood & Tissue Genomic Mini )………………………………………………………….29
4、 引子組SEN1/2、SEO1/2、SEP1/2之PCR特異性試驗……..29
5、 引子組SEN1/2、SEO1/2、SEP1/2之PCR靈敏度試驗……..30
(1) 直接檢測……………………………………………………….30
(2) 二次活化……………………………………………………….30
(三) SEN、SEO、SEP特異性引子組於食品檢測之應用………………31
1、 豬肉樣品之檢測應用…………………………………………31
(1) 豬肉中生菌數之計算………………………………………….31
(2) 肉品中凝固正反應之葡萄球菌總菌數……………………32
(3) 檢測之靈敏度………………………………………………….32
2、 鮮乳樣品的檢測應用………………………………………..32
(1) 鮮乳樣品中生菌數與S. aureus之計算………………………..32
(2) 菌培養……………………………………………………….32
(四) 反轉錄聚合鏈鎖反應 ( Reverse transcription-Polymerase Chain Reaction; RT-PCR )…………………………………………………33
(五) 金黃色葡萄球菌之腸毒素之分布情形…………………………….33
參、結果與討論……………………………………………………………….34
金黃色葡萄球菌新型腸毒素N、O、P特異性引子組之設計與應用
一、標準菌的選定與PCR引子組之設計…………………………………34
二、聚合鏈鎖反應 ( PCR )……………………………………………..35
(一) PCR引子組特異性試驗…………………………………………..35
(二) PCR引子組靈敏度試驗………………………………………......35
三、反轉錄聚合鏈鎖反應 ( Reverse transcription PCR )……………...36
金黃色葡萄球菌腸毒素之分佈
一、 中毒型休克症候群毒素(TSST-1)基因之分佈情形…………………38
二、 新型腸毒素基因之分佈……………………………………………...38
三、 中毒性休克症候群毒素及新型腸毒素基因之相關性與分布關係...38
肆、結論……………………………………………………………………….42
Ⅲ、參考文獻……………………………………………………………….......77








表 次
頁次
表1-1、民國70年至93年台灣地區食品中毒案件病因物質分類表................18
表1-2、金黃色葡萄球菌相關的超級抗原位置…………………………………19
表1-3、金黃色葡萄球菌各腸毒素胺基酸序列相似性之比較………………….20
表1-4、金黃色葡萄球菌腸毒素之主要特性…………………………………….21
表2-1、本實驗用於篩選實驗室標準菌株之腸毒素基因之PCR引子組……….44
表2-2、本實驗所使用之PCR引子組…………………………………………….45
表2-3、本研究所使用之PCR反應條件.................................................................46
表2-4、本研究所使用之非金黃色葡萄球菌葡萄球菌屬標準菌株………………47
表2-5、金黃色葡萄球菌腸毒素標準菌…………………………………………….48
表2-6、金黃色葡萄球菌腸毒素標準菌偵測sen,seo和sep腸毒素基因…………49
表2-7、以PCR檢測腸毒素型S. aureus之靈敏度……………………………….50
表2-8、以PCR技術檢測1992-1993年台中榮總臨床檢體分離之金黃色葡萄球菌腸毒素型基因布………………………………………………………………….51
表2-9、以PCR技術檢測2003年台中榮總臨床檢體分離之金黃色葡萄球菌腸毒素型基因分布……………………………………………………………………….53
表2-10、以PCR技術檢測2003年疾病管制局食品中毒案件臨床檢體球菌分離之金黃色葡萄腸毒素型基因布……………………………………………………55
表2-11、PCR引子組檢測金黃色葡萄球菌臨床分離株腸毒素基因之分布……61表2-12、腸毒素基因sen, seo and sep與傳統型毒素基因(sea~see)之分佈…….62
圖 示
頁次
圖2-1、利用 (a)SEN1/SEN2,(b)SEO1/SEO2及(c)SEP1/SEP2引子組對實驗室標準菌及非金黃色葡萄球菌之PCR增幅作用…………………………………….63
圖2-2、SEN1/SEN2引子組對實驗室標準菌株STA67之靈敏度……………..64
圖2-3、SEO1/SEO2引子組對實驗室標準菌株STA34之靈敏度……………..65
圖2-4、SEP1/SEP2引子組對實驗室標準菌株HYT59之靈敏度……………..66
圖2-5、SEN1/SEN2引子組檢測牛乳及豬肉中腸毒素sen基因之靈敏度……67
圖2-6、SEO1/SEO2引子組檢測牛乳及豬肉中腸毒素seo基因之靈敏度……68
圖2-7、SEP1/SEP2引子組檢測牛乳及豬肉中腸毒素sep基因之靈敏度……..69
圖2-8、利用(a)SEN1/SEN2, (b)SEO1/SEO2及(c)SEP1/SEP2引子組對金黃色葡萄球菌之RT-PCR增幅作用...................................................................................70
圖 2-9、定序結果 (STA67)與N型腸毒素基因序列之比對...............................71
圖 2-10、定序結果 (STA34)與O型腸毒素基因序列之比對………………….72
圖 2-11、定序結果 (HYT59)與P型腸毒素基因序列之比對………………….73
圖 2-12、PCR結果與N型腸毒素基因序列之比對……………………………..74
圖 2-13、PCR結果與O型腸毒素基因序列之比對……………………………..75
圖 2-14、PCR結果與P型腸毒素基因序列之比對……………………………..76

張長泉。1996。金黃色葡萄球菌快速檢測之最近發展狀況。食品工業月刊，40-49。
張立同Chang, L. T. 2004。金黃色葡萄球菌新型腸毒素K、L、M基因引子之設計及其應用於食品及臨床菌株腸毒素型之分布調查。中興大學食品科學研究所碩士論文。
Abigail, A. S. and D. D. Whitt. 2001. Bacterial Pathogenesis: a Molecular Approach, Second Edition. P. 216-231. ASM Press.
Bayles K. W and J. J. Iandolo. 1989. Genetic and molecular analyses of the gene encoding staphylococcal enterotoxin D. J. Bacteriol. 171: 4799-4806.
Becker K, A. W. Friedrich, G. Peters and C. von Eiff. 2004. Systematic survey on the prevalence of genes coding for staphylococcal enterotoxins SElM, SElO, and SElN. Mol. Nutr. Food Res. 48(7):488-95.
Bennett, R. W., and Lancette, G. A. 1992. Staphylococcus aureus, pp. 161-166. In Bacteriological Analytical Manual, 7th ed. Association Official Analytical Chemists International, Arlington, VA..
Bergdoll, M. S., M. J. Surgalla, and G. M. Dack. 1959. Identification of staphylococcal enterotoxin B. J. Immunol. 83: 334-338.
Bergdoll, M. S., E. J. Schantz, W. G. Roellser, J. Wagman, L. Spero and D. A. Dunnery. 1965. Purification of staphylococcal enterotoxin B. Biochemistry. 4(6):1011-6.
Bergdoll, M. S., 1967. The staphylococcal enterotoxins. In: Mateles, R. I., Wogan, G. N. (ed.), Biochemistry of Some Foodborne Microbial Toxins. MIT Press, Cambridge, pp. 1– 25.
Bergdoll, M. S., C. R. Borja, R. N. Robbins, and K. F. Weiss. 1971. Identification of staphylococcal enterotoxin E. Infect. Immunol. 4: 593-595.
Bergdoll, M. S. 1983. Enterotoxins, p.559-598. In C., Adlam, and C. S. F., Easmon (ed). Staphylococci and staphylococcal infections, Vol. 2. Academic press, Inc., London.
Betley, M. J., S. Lofdahl, B. N. Kreiswirth, M. S. Bergdoll, and R. P. Novick. 1984. Staphylococcal enterotoxin A gene is associated with a variable genetic element. Proc. Natl. Acad. Sci. USA. 81: 5197-5183.
Betley, M. J., and J. J. Mekalanos. 1985. Staphylococcal enterotoxin A is encoded by phage. Science. 12;229(4709):185-7.
Betley, M. J., and J. J. Mekalanos. 1988. Nucleotide sequence of the type A staphylococcal enterotoxin gene. J. Bacteriol. 170: 34-41.
Betley, M. J., D. W. Borst, and L. B. Regassa. 1992. Staphylococcal enterotoxins, toxic shock syndrome toxin and streptococcal pyrogenic exotoxins: a comparative study of their molecular biology. Chem. Immunol. 55: 1-35.
Betley, M. J., and T. O. Harris. 1994. Staphylococcal enterotoxins: genetic characterization and relationship between structure and emetic activity. J. Food Microbiol. 11: 109-121.
Blaiotta, G., D. Ercolini, C. Pennacchia, V. Fusco, A. Casaburi, O. Pepe and F. Villani. 2004. PCR detection of staphylococcal enterotoxin genes in Staphylococcus spp. strains isolated from meat and dairy products. Evidence for new variants of seG and seI in S. aureus AB-8802. J. Appl. Microbiol. 97(4):719-30.
Bohach, G. A., and P. M. Schlievert. 1987. Nucleotide sequence of the staphylococcal enterotoxin C1 gene and relatedness to other pyrogenic toxins. Mol. Gen. Genet. 209: 15-20.
Bohach, G. A., and P. M. Schlievert. 1989. Conservation of the biologically active portions of staphylococcal enterotoxins C1 and C2. Infect. Immunol. 57: 2249-2252.
Borja, C. R., and M. S. Bergdoll. 1967. Purification and partial characterization of enterotoxin C produced by Staphylococcus aureus strain 137. Biochemistry. 6: 1467-1473.
Borja, C. R., E. Fenning, I. Y. Huang, and M. S. Bergdoll. 1972. Purification and some physicochemical properties of staphylococcal enterotoxin E. J. Biol. Chem. 247: 2456-2463.
Borst, D. W., and Betley M. J. 1993. Mutations in the promoter spacer region and early transcribed region increase expression of staphylococcal enterotoxin A. Infect. Immunol. 61: 5421-5.
Borst, D. W. and M. J. Betley. 1994. Phage-associated differences in staphylococcal enterotoxin A gene (sea) expression correlate with sea allele class. Infect Immunol. 1994 Jan;62(1):113-8.
Brakstad O. G., K. Aasbakk and J. A. Maeland . 1992. Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene. J. Clin. Microbiol. 30: 1654-60.
Casman, E. P. 1960. Identification of staphylococcal enterotoxin A. J. Bacteriol. 79: 849-856.
Casman, E. P., R. W. Bennett, A. E. Dorsey, and J. A. Issa. 1967. Identification of a fourth staphylococcal enterotoxin, enterotoxin D. J. Bacteriol. 94: 1875-1882.
Chang, H. C., and M. S. Bergdoll. 1979. Purification and some physiochemical properties of staphylococcal enterotoxin D. Biochemistry. 18: 1937-1942.
Chu, F. S., K-Thadhani, E. J. Schantz, and M. S. Bergdoll. 1966. Purification and characterization of staphylococcal enterotoxin A. Biochemistry. 5: 3281-3289.
Couch, J. L., M. T. Soltis, and M. J. Betley. 1988. Cloning and nucleotide sequence of the type E staphylococcal enterotixn gene. J. Bacteriol. 170: 2954-2960.
Couch, J. L., and M. J. Betley. 1989. Nucleotide sequence of the type C3 staphylococcal enterotoxin gene suggests that intergenic recombination causes antigenic variation. J. Bacteriol. 171: 4507-4510.
Crass, B. A., and M. S. Bergdoll. 1986. Involvement of staphylococcal enterotoxin in nonmenstrual toxic shock syndrome. J. Clin. Microbiol. 23: 1138-1139.
Czop, J. K., and M. S. Bergdoll. 1974. Staphylococcal enterotoxin synthesis during the exponential, transitional, and stationary growth phases. Infect. Immunol. 9: 229-235.
De Boer, M. L. and A. W. Chow. (1994) Toxic shock syndrome toxin 1—producing Staphylococcus aureus isolates contain the staphylococcal enterotoxin B genetic element but do not express staphylococcal enterotoxin B. J. Infect. Dis. 170, 818–827.
Dinges, M. M., Orwin, P. M., Schlievert, P. M. 2000. Exotoxins of Staphylococcus aureus. Clin. Microbiol. Rev. 13:16-34.
Elek, S. D. 1959. Staphylococcus aureus and its relation to disease. Churchill Livingstone, New York.
Evenson, M. L., Hinds, M.W., Bernstein, R. S., Bergdoll, M. S. 1988. Estimation of human dose of staphylococcal enterotoxin A from a large outbreak of staphylococcal food poisoning involving chocolate milk. Int. J. Food Microbiol. 7: 311–6.
Fitzgerald, J. R., S. R. Monday, T. J. Foster, G. A. Bohach, P. J. Hartigan, W. J. Meaney, and C. J. Smyth. 2001. Characterization of a putative pathogenicity island from bovine Staphylococcus aureus encoding multiple superantigens. J. Bacteriol. 183: 63-70.
Freed, R. C., M. L. Evenson, R. F. Reiser, and M. S. Bergdoll. 1982. Enzyme-linked immunosorbent assay for detection of staphylococcal enterotoxins in foods. Appl. Environ. Microbiol. 44: 1349-1355.
Fueyo J. M., M. C. Mendoza, M. R. Rodicio, J. Muiz, M. A. Alvarez, and M. C. Martn. 2005. Cytotoxin and pyrogenic toxin superantigen gene profiles of staphylococcus aureus associated with subclinical mastitis in dairy cows and relationships with macrorestriction genomic profiles. J. Clin. Microbiol. 43(3): 1278–1284.
Fung-Tomo, J. H., Radelski, E. G., Denbleyker, K., Bonner, D. P. and Kessler, R. E. 1991. Emergence of homogeneously methicillin-resistant Staphylococcus aureus. J. Clin. Microbiol. 29: 2880-3.
Genigeorgis, C. A. 1989. Present state of knowledge on staphylococcal intoxication. Int. J. Food Microbiol. 9: 327-360.
Garbe, P. L., R. J. Arko, and A. L. Reingold. 1985. Staphylococcus aureus isolates from patients with nonmenstrual TSS. JAMA. 253: 2538-2542.
Gaskill, M. E., and S. A. Khan. 1988. Regulation of the enterotoxin B gene in Staphylococcus aureus. J. Biol. Chem. 263: 6276-6280.
Holtfreter, S., K. Bauer, D. Thomas, C. Feig, V. Lorenz, K. Roschack, E. Friebe, K. Selleng, S. Lvenich, T. Greve, A. Greinacher, B. Panzig, S. Engelmann, G. Lina, and B. M. Brker. 2004. egc-Encoded Superantigens from Staphylococcus aureus Are Neutralized by Human Sera Much Less Efficiently than Are Classical Staphylococcal Enterotoxins or Toxic Shock Syndrome Toxin. Infect. Immunol. 72(7): 4061–4071.
Hoover, D. G., S. R. Tatini and J. B. Maltais. 1983. Characterization of staphylococci. Appl. Environ. Microbiol. 46(3): 649–660.
Hovde, C. J., J. C. Marr, M. L. Hoffmann, S. P. Hackett, Y. I. Chi, K. K. Crum,D. L. Stevents,C. V. Stauffacher, and G. A. Bohach. 1994. Investigation of the role of the disulphide bond in the activity and structure of staphylococcal enterotoxin C1. Mol. Microbiol. 13: 897-909.
Hovde, C. J., S. P. Hackett, and G. A. Bohach. 1990. Nucleotide sequence of the staphylococcal enterotoxin C3 gene : sequence comparison of all three type C staphylococcal enterotoxins. Mol. Gen. Genet. 220: 329-333.
Hu, D. L., K. Omoe , A. Nakane, K. Shinagawa and H. T. Omoe. 2003. Identification and characterization of a new staphylococcal enterotoxin-related putative toxin encoded by two kinds of plasmids. Infect. Immunol. 71: 6088-6094.
Huang, I. Y., T. Shih, C. R. Borja, R. M. Avena, and M. S. Bergdoll. 1967. Amino acid composition and terminal amino acids of staphylococcal enterotoxin C. Biochemistry. 6: 1480-1483.
Huang, I. Y., and M. S. Bergdoll. 1970. The primary structure of staphylococcal enterotoxin B. J. Biol. Chem. 245: 3518-3525.
Huang, I. Y., J. L. Hughes, M. S. Bergdoll, and E. J. Schantz. 1987. Complete amino acid sequence of staphylococcal enterotoxin A. J. Biol. Chem. 262: 7006-7013.
Iandolo, J. J. 1989. Genetic analysis of extracellular toxins of Staphylococcus aureus. Annu. Rev. Microbiol. 43: 375-402.
Jarraud, S., G. Cozon, F. Vandenesch, M. Bes, J. Etienne and G. Lina. 1999. Involvement of Enterotoxins G and I in Staphylococcal Toxic Shock Syndrome and Staphylococcal Scarlet Fever. J. Clin. Microbiol. 37. (8):2446-9.
Jarraud, S., M. A. Peyrat, A. Lim, A. Tristan, M. Bes, C. Mougel, J. Etienne, F. Vandenesch, M. Bonneville, and G. Lina. 2001. egc, A highly prevalent operon of enterotoxin gene, forms a putative nursery of superantigens in Staphylococcus aureus. J. Immunol. 166: 669-677.
Johnson, W. M., S. D. Tyler, E. P. Ewan, F. E. Ashton, D. R. Pollard, and K. R. Rozee. 1991 Detection of genes for enterotoxins, exfoliative toxins and toxic shock syndrome toxin 1 in Staphylococcus aureus by the polymerase chain reaction. J. Clin. Microbiol. 29, 426–430.
Kloos, W. E. and K. H. Schleifer. 1975. Simplified scheme for routine identification of human Staphylococcus species. J. Clin. Microbiol. 1(1): 82–88.
Kokan, N. P., and M. S. Bergdoll. 1987. Detecting of low-enterotoxin-producing Staphylococcus aureus strains. Appl. Environ. Microbiol. 53: 2675-2676.
Kuroda, M., T. Ohta, I. Uchiyama, T. Baba, H. Yuzawa, I. Kobayashi, L. Cui, A. Oguchi, K. Aoki, Y. Nagai, J. Lian, T. Ito, M. Kanamori, H. Matsumaru, A. Maruyama, H. Murakami, A. Hosoyama, Y. Mizutani-Ui, N. K. Takahashi, T. Sawano, R. Inoue, C. Kaito, K. Sekimizu, H. Hirakawa, S. Kuhara, S. Goto, J. Yabuzaki, M. Kanehisa, A. Yamashita, K. Oshima, K. Furuya, C. Yoshino, T. Shiba, M. Hattori, N. Ogasawara, H. Hayashi, and K. Hiramatsu. 2001. Whole genome sequencing of methicillin-resistant Staphylococcus aureus. Lancet 357:1225–1240.
Lam, T. J., A. Pengov, A. Schukken, Y. H. Smit, and A. Brand. 1995. The differentiation of Staphylococcus aureus from other Micrococcaceae isolated from bovine mammary glands. J. Appl. Bacteriol. 79:69-72.
Lancette, G. A., and Tatini, S. R., 1990. Staphylococcus aureus, pp. 533-50. In C. Vanderzant and D. F. Splittstoesser(ed.), Compendium of Methods for the Microbiological Examination of Foods, 3rd ed. American Public Health Association, Washington, DC.
Letertre, C., Perelle, S., Dilasser, F. and Fach, P. 2003. Identification of a new putative enterotoxin SEU encoded by the egc cluster of Staphylococcus aureus. J. Appl. Microbiol. 95: 38-43.
Le Loir Y, F. Baron and M. Gautier. 2003. Staphylococcus aureus and food poisoning. Genet. Mol. Res. 31;2(1):63-76.
Lina, G., G.A. Bohach, , S. P. Nair, K. Hiramatsu, E. Jouvin-Marche, and R. Mariuzza. 2004. Standard nomenclature for the superantigens expressed by Staphylococcus. J. Infect. Dis. 189, 2334–2336.
Mackay, A. D., A. Quick, S. H. Gillespie, and C. C. Kibbler. 1993. Coagulase-negative methicillin-resistant Staphylococcus aureus infection. Lancet. 342(8869): 492.
Mallonee, D. H., B. A. Glatz, and P. A. Pattee. 1982. Chromosomal mapping of a gene affecting enterotoxin A production in Staphylococcus aureus. Appl. Environ. Microbiol. 43: 397-402.
Marr, J, C., J. D. Lyon, J. R. Roberson, M. Lupher, W. C. Davis, and G. A. Bohach. 1993. Characterization of novel type C staphylococcal enterotoxins: biological and evolutionary implications. Infect. Immunol. 61: 4254-4262.
Marrack, P., and J. Kappler. 1990. The staphylococcal enterotoxins and their relatives. Science. 248: 705-711.
McLauchlin, J., G. L. Narayanan., V. Mithani., G. O’Neill., 2000. The detection of enterotoxins and toxic shock syndrome toxin genes in Staphylococcus aureus by polymerase chain reaction. J. Food Prot. 63: 479-488
Mempel,M., G. Lina, M. Hojka, C. Schnopp, H.-P. Seidl, T. Schfer, J. Ring, F. Vandenesch and D. Abeck. 2003. High Prevalence of Superantigens Associated with the egc Locus in Staphylococcus aureus Isolates from Patients with Atopic Eczema. Eur. J. Clin. Microbiol. Infect. Dis. 22(5):306-9.
Miller, B. A., R. F. Reiser, and M. S. Bergdoll. 1978. Detection of staphylococcal enterotoxins A, B, C, D, and E in foods by radioimmunoassay, using staphylococcal cells containing protein A as immunoadsorbent. Appl. Environ. Microbiol. 36: 421-426.
Munson, S. H., and M. J. Betley. 1991. Partial characterization of a new staphylococcal enterotoxin gene, abstr. B-36, p.31. In Abstracts of the 91st General meeting of the American Society for Microbiology 1991. American Society for Microbiology, Washington, D.C.
Munson, S. H., M. T. Tremaine, M. J. Betley, and R. A. Welch. 1998. Identification and characterization of Staphylococcal enterotoxin types G and I from Staphylococcus aureus. Infect. Immunol. 66: 3337-3348.
Murono, K., Fujita, K. and Yoshioka, H. 1988. Detection of staphylococcal exfoliative toxin by slide latex agglutination. J. Clin. Microbiol. 26: 271-4.
Novick, R. P., P. Schlievert and A. Ruzin.2001. Pathogenicity and resistance islands of staphylococci. Microbes Infect. 3: 585-594.
Omoe, K., M. Ishikawa, Y. Shimoda, D. L. Hu, S. Ueda, and K. Shinagawa. 2002. Detection of seg, seh, and sei genes in Staphylococcus aureus isolates and determination of the enterotoxin productivities of S. aureus isolates harboring seg, seh, or sei genes. J. Clin. Microbiol. 40:857-862.
Omoe, K., D. L. Hu, H. T. Omoe, A. Nakane, and K. Shinagawa. 2003. Identification and characterization of a new staphylococcal enterotoxin-related putative toxin encoded by two kinds of plasmids. Infect. Immunol. 71: 6088-6094.
Omoe, K., K. Imanishi, D. L. Hu, H. Kato, H. T. Omoe, A. Nakane, T. Uchiyama and K. Shinagawa. 2004. Biological Properties of Staphylococcal Enterotoxin-Like Toxin Type R. Infect. Immunol. 72(6): 3664–3667.
Omoe, K., K. Imanishi, D. L. Hu, H. Kato, Y. Fugane, Y. Abe, S. Hamaoka, Y. Watanabe, A. Nakane, T. Uchiyama and K. Shinagawa. 2005. Characterization of novel staphylococcal enterotoxin-like toxin type P. Infect. Immunol. 73:5540-5546.
Orwin, P. M., D. Y. M. Leung, H. L. Donahue, R. P. Novick, and P. M. Schlievert. 2001. Biochemical and biological properties of staphylococcal enterotoxin K. Infect. Immunol. 69: 360-366.
Orwin, P. M., D. Y. M. Leung, T. J. Tripp, G. A. Bohach, C. A. Earhart, D. H. Ohlendorf, and P. M. Schlievert. 2002. Characterization of a novel staphylococcal enterotoxin-like superantigen, a member of the group V subfamily of pyrogenic toxins. Biochemistry. 41: 14033-14040.
Orwin, P. M., J. R. Fitzgerald, D. Y. M. Leung, L. A. Gutierrez, G. A. Bohach, and P. M. Schlievert. 2003. Characterization of Staphylococcus aureus enterotoxin L. Infect. Immunol. 71: 2916-2919.
Otero, A., M. L. Garcia, M. C. Farcia, B. Moreno, and M. S. Bergdoll. 1990. Production of staphylococcal enterotoxins C1 and C2 and thermonuclease throughout the growth cycle. Appl. Environ. Microbiol. 56: 555-559.
O''Toole, P. W., and Foster, T. J. 1986. Molecular cloning and expression of epidermolytic toxin A gene of Staphylococcus aureus Microb. Pathog. 1: 583-94.
Park, C. E., and R. Szabo. 1986. Evaluation of the reversed passive latex agglutination (RPLA) test kits for detection of staphylococcal enterotoxins A, B, C, and D in foods. Can. J. Microbiol. 32: 723-727.
Pattee, P. A., and B. A. Glatz. 1980. Identification of a chromosomal determinant of enterotoxin A production in Staphylococcus aureus. Appl. Environ. Microbiol. 39: 186-193.
Recsei, P., B. Kreiswirth, M. O’Reilly, P. Schlievert, A. Gruss, and R. P. Novick. 1986. Regulation of exoprotein gene expression in Staphylococcus aureus by agr. Mol. Gen. Genet. 202: 58-61.
Regassa, L. B., J. L. Couch, and M. J. Betley. 1991. Steady-state staphylococcal enterotoxin type C mRNA is affected by a product of the accessory gene regulator (agr) and by glucose. Infect. Immunol. 59: 955-962.
Reiser, R. F., R. N. Robbins, A. Noleto, G. P. Khoe, and M. S. Bergdoll. 1984. Identification, purification, and some physicochemical properties of staphylococcal enterotoxin C3. Infect. Immunol. 45: 625-630.
Ren, K., J. D. Bannan, V. Pancholi, A. L. Cheung, J. C. Robbins, V. A. Fischetti, J. B. Zabriskie. 1994. Characterization and biological properties of a new staphylococcal exotoxin. J. Exp. Med. 1;180(5):1675-83.
Richard, P. N., P. Schlievert, and A. Ruzin. 2001. Pathogenicity and resistance islands of staphylococci. Microbe. Infect. 3: 585-594.
Roberson, J. R., L. K. Fox, D. D. Hancock, and T. E. Besser. 1992. Evaluation of methods for the differentiation of coagulase-positive staphylococci. J. Clin. Microbiol. 30:3217-3219.
Robbins, R., S. Gould., and M. S. Bergdoll. 1974. Detecting the enterotoxigenicity of Staphylococcus aureus strains. Appl. Microbiol. 28: 946-950.
Rosec J. P., O. Gigaud. 2002. Staphylococcal enterotoxin genes of classical and new types detected by PCR in France. Int. J. Food microbiol. 77: 61-70.
Rosenblum, E. D. and S. Tyrone. 1976. Chromosomal determinants for exfoliative toxin production in two strains of staphylococci. Infect. Immunol. 14(5): 1259–1260.
Schantz, E. J., J. Wagman and R. C. Edwards. 1965. Molecular size, homogeneity, and hydrodynamic properties of purified staphylococcal enterotoxin B. Biochemistry. 4(6):1017-23.
Schantz, E. J., W. G. Rossler, M. J. Woodburn, J. M. Lyneh, H. M. Jacoby, S. J. Silverman, J. C. Gorman, and L. Spero. 1972. Purification and some chemical and physical properties of staphylococcal enterotoxin A. Biochemistry. 11: 860-866.
Schlievert, P. M. 1986. Staphylococcal enterotoxin B and toxic shock syndrome toxin 1 are significantly associated with non-menstrual TSS. Lancet. 1: 1149-1150.
Schmidt, J. J., and L. Spero. 1983. The complete amino acid sequence of staphylococcal enterotoxin C1. J. Biol. Chem. 258: 6300-6306.
Schwalbe, R. S., Stapleton, J. T. and Gilligan, P. H. 1987. Emergence of vancomycin resistance in coagulase-negative staphylococci. N. Engl. J. Med. 316: 927-31.
Schwalbe, R. S., Ritz, W. J., Verma, P. R., Barranco, E. A., and Gilligan, P. H. 1990. Selection for vancomycin resistance in clinical isolates of Staphylococcus haemolyticus. J. Infect. Dis. 161: 45-51.
Shafer, W. M., and J. J. Iandolo. 1979. Genetics of staphylococcal enterotoxin B in methicillin resistant isolates of Staphylococcus aureus. Infect. Immunol. 25: 902-911.
Smeltzer, M. S., M. E. Hart, and J. J. Iandolo. 1993. Phenotypic characterization of xpr, a global regulator of extracellular virulence factors in Staphylococcus aureus. Infect. Immunol. 61: 919-925.
Sundstrom, M., Abrahmsen, L., Antonsson, P., Mehindate, K., Mourad, W., and Dohlsten, M. 1996. The crystal structure of staphylococcal enterotoxin type D reveals Zn2+-mediated homodimerization. EMBO J. 15: 6832-40.
Su, Y. C., and A. C. L. Wong. 1993. Optimal condition for the production of unidentified staphylococcal enterotoxins. J. Food Prot. 56: 313-316.
Su, Y. C., and A. C. L. Wong. 1995. Identification and purification of a new staphylococcal enterotoxin, H. Appl. Environ. Microbiol. 61: 1438-1443.
Smyth, D. S., P. J. Hartigan, W. J. Meaney, J. R. Fitzgerald, C. F. Deobald, G. A. Bohach and C. J. Smyth. 2005. Superantigen genes encoded by the egc cluster and SaPIbov are predominant among Staphylococcus aureus isolates from cows, goats, sheep, rabbits and poultry. J. Med. Microbiol. 54: 401-411.
Tremaine, M. T., D. B. Brockman, and M. J. Betley. 1993. Staphylococcal enterotoxin A gene (sea) expression is not affected by the accessory gene regulator (agr). Infect. Immunol. 61: 356-359.
Tsen, H. Y., T. R. Chen and G. K. Yu. 1994. Detection of B, C types enterotoxigenic Staphylococcus aureus using polymerase chain reaction. J. Chinese Agric. Chem. Soci. 3: 322-331.
Udo, E. E., M. A. Al-Bustan, L. E. Jacob, and T. D. Chugh. 1999. Enterotoxin production by coagulase-negative staphylococci in restaurant workers from Kuwait City may be a potential cause of food poisoning. J. Med. Microbiol. 48(9):819-23.
Vandenesch, F., C. Lebeau, M. Bes, G. Lina, B. Lina, T. Greenland, Y. Benito, Y. Brun, Fleurette J., and J. Etienne. 1994. Clotting activity in Staphylococcus schleiferi subspecies from human patients. J. Clin. Microbiol. 32:388–392.
Wang, S. J., L. W. Chow and L. J. Wu. 2002. Multiplex PCR for the simultaneous detection of the SEA, SEB, SEC, SED and SEE genes of enterotoxigenic Staphylococcus aureus. J. Food. Drug Anal. 10(3): 164~169.
Warren, R., Rogolsky, M., Wiley, B. B. and Glasgow, L. A. 1974. Effect of ethidium bromide on elimination of exfoliative toxin and bacteriocin production in Staphylococcus aureus. J. Bacteriol. 118: 980-985.
Warren, R., Rogolsky, M., Wiley, B. B. and Glasgow, L. A. 1975. Isolation of extrachromosomal deoxyribonucleic acid for exfoliative toxin production from phage group 11 Staphylococcus aureus. J. Bacteriol. 122: 99-105.
Williams, R. J., J. M. Ward, B. Henderson, S. Poole, B. O’Hara, M. Wilson and S. Nair. 2000. Identification of a novel gene cluster encoding staphylococcal enterotoxin-like proteins: characterization of the prototypic gene and its protein product, SET1. Infect. Immunol. 68(8): 4407-15.
Yarwood J. M., J. K. McCormick, M. L. Paustian, P. M. Orwin, V. Kapur, and P. M. Schlievert. 2002. Characterization and expression analysis of Staphylococcus aureus pathogenicity island 3. J. Biol. Chem. 277: 13138-13147.
Zhang, S., J. J. Iandolo, and G. C. Stewart. 1998. The enterotoxin D plasmid of Staphylococcus aureus encodes a second enterotoxin determinant (sej). FEMS Microbiol. Lett. 15;168(2): 227-33.