Application of a Chimeric Protein Construct having Enterotoxin B and Toxic Shock Syndrome Toxin Domains of S. aureus in Immunodiagnostics
详细信息 查看全文
- 作者：R. Shylaja (1)
Devi Kalyan Kumar Thakasi (1)
H. S. Murali (1)
K. Prakash Narayana Reddy (1)
H. V. Batra (1) - 关键词：Seb ; Tsst ; Chimeric protein ; S. aureus r ; TE ; Superantigens ; Western blot
- 刊名：Indian Journal of Microbiology
- 出版年：2012
- 出版时间：September 2012
- 年：2012
- 卷：52
- 期：3
- 页码：449-455
- 全文大小：336KB
- 参考文献：1. Kamboj DV, Nema V, Pandey AK, Goel AK, Singh L (2006) Heterologous expression of staphylococcal enterotoxin B ( / seb) gene for antibody production. Electron J Biotechnol ISSN: 0717-3458. 9(5)
2. Mehrotra M, Wang G, Wendy JM (2000) Multiplex PCR for detection of genes for / Staphylococcus aureus enterotoxins, exfoliative toxins, toxic shock syndrome toxin 1, and methicillin resistance. J Clin Microbiol 38(3):1032-035
3. Blanco L, Choi EM, Connolly K, Thompson MR, Bonventre PF (1990) Mutants of staphylococcal toxic shock syndrome toxin 1: mitogenicity and recognition by a neutralizing monoclonal antibody. Infect Immun 58(9):3020-028
4. De-Lourdes Maria RS, Da-Cunha RAO, Junior CJP (2007) Detection of enterotoxin, toxic shock syndrome toxin 1 genes in / Staphylococcus with emphasis on coagulase negative / Staphylococci. Microbiol Immunol 51(4):381-90
5. Kansal R, Davis C, Hansmann M, Seymour J, Parsonnet J, Modern P, Gilbert S, Kotb M (2007) Structural and functional properties of antibodies to the superantigen TSST-1 and their relationship to menstrual toxic shock syndrome. J Clin Immunol 27(3):327-38 CrossRef
6. Rajagopalan G, Sen MM, Singh M, Murali NS, Nath KA, Iijima K, Kita H, Leontovich AA, Gopinathan U (2006) Intranasal exposure to staphylococcal enterotoxin B elicits an acute systemic inflammatory response. Shock 25:647-56 CrossRef
7. Anderson MR, Tary-Lehmann M (2001) Staphylococcal enterotoxin-B-induced lethal shock in mice is T-cell-dependent, but disease susceptibility is defined by the non-T-cell compartment. Clin Immunol 98:85-4 CrossRef
8. Hemalatha V, Srikanth P, Mallika M (2004) Superantigens: concepts, clinical disease and therapy. Indian J Med Microbiol 22(4):204-11
9. Konno O, Hirano T, Katsuyama K, Oka K, Matsuno N, Nagao T (2007) Bacterial superantigen TSST-1 attenuates suppressive efficacy of glucocorticoids and calcineurin inhibitors against blastogenesis of peripheral blood mononuclear cells from patients with chronic renal failure on hemodialysis treatment. Transpl Immunol 17(3):187-92 CrossRef
10. Ulrich RG, Sidell S, Taylor TJ, Wilhelmsen CL, Franz DR (1997) Staphylococcal enterotoxin B and related pyrogenic toxins. In: Sidell FR, Takafuji ET, Franz DR (eds) Textbook of military medicine Part I, Warfare, weaponry, and the casualty: medical aspects of chemical and biological warfare. Washington, DC: Office of the Surgeon General, Department of the Army, United States of America, pp 621-30
11. Kumar TDK, Balakrishna K, Murali HS, Batra HV (2009) Construction of a recombinant intergenus multidomain chimeric protein for simultaneous expression of haemolysin BL of / Bacillus cereus, listeriolysin O of / Listeria monocytogenes and enterotoxin B of / Staphylococcus aureus. J Med Microbiol 58:577-83 CrossRef
12. Sambrook J, Fritsch E, Maniatis A (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor
13. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin–phenol reagents. J Biol Chem 193:265-75
14. Engvall E, Perlman P (1971) Enzyme-linked immunosorbent assay: quantitative assay of immunoglobulin G. Immunochemistry 8:871-74 CrossRef
15. Harlow E, Lane D (1988) Antibodies: a laboratory manual. CSHL Press, Cold Spring Harbor
16. Nadig S, Ramachandra Raju S, Arakere G (2010) Epidemic meticillin-resistant / Staphylococcus aureus (EMRSA-15) variants detected in healthy and diseased individuals in India. J Med Microbiol 59(7):815-21 CrossRef
17. Chini V, Dimitracopoulos G, Spiliopoulou I (2006) Occurrence of the enterotoxin gene cluster and the toxic shock syndrome toxin 1 gene among clinical isolates of MRSA is related to clonal type and / agr group. J Clin Microbiol 44(5):1881-883 CrossRef - 作者单位：R. Shylaja (1)
Devi Kalyan Kumar Thakasi (1)
H. S. Murali (1)
K. Prakash Narayana Reddy (1)
H. V. Batra (1)
1. Department of Microbiology, Defence Food Research Laboratory, Siddharthanagar, Mysore, 570011, India - ISSN：0973-7715
文摘
Staphylococcal enterotoxin B (SEB) and toxic shock syndrome toxin-1 are the super antigens responsible for diseases such as staphylococcal food poisoning and toxic shock syndrome. At low serum concentrations, SEB can trigger toxic shock, profound hypotension and multi organ failure and hence is recognized as biowarfare molecule. In this study, a multidomain fusion protein (r-TE) was generated with specificity for SEB and toxic shock syndrome toxin (Tsst-1). The fusion gene comprising the conserved regions of seb and the tsst genes was codon-optimized for expression in Escherichia coli and encoded a 26?kDa recombinant multidomain chimeric protein (r-TE). Hyperimmune antiserum raised against r-TE specifically reacted with SEB (~28?kDa) and Tsst-1 (~22?kDa) components during Western blot analysis and by plate ELISA in confirmed toxin producing strains of S. aureus. The antigenicity of the SEB component of the r-TE protein was also confirmed using TECRA kit. The described procedure of creating a single protein molecule carrying components of two different toxins whilst still retaining the original antigenic determinants of individual toxins proved highly advantageous in the development of rapid, reliable and cost effective immunoassays and may also have the potential to serve as candidate molecule for vaccine studies.