candidate genes for vaccine in Methicillin-resistant ... - Semantic Scholar

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Kowsar Medical Journal ... Ù­Department of Microbiology, Faculty of Health, Tehran University of Medical Sciences ... 2MARS Bioinformatics Institute, Tehran, Iran.
Kowsar Medical Journal Vol. 16, No. 3, Fall 2011 Pages: 145-149

saCOL2291 saCOL2581

PhD

*

PhD PhD

MSc PhD *

saCOL2581 saCOL2291

CSF PCR

DNA

Gene runner

Puls4

N C C saCOL

Gene polymorphism of saCOL2291 and saCOL2581; candidate genes for vaccine in Methicillin-resistant Staphylococcus aureus Rouhani Nejad H.1 MSc, Fallah Mehrabadi J.2 PhD, Pourmand M. R.* PhD, Zolfaghari M. R.1 PhD, Imani Fooladi A. A.3 PhD ‫٭‬Department of Microbiology, Faculty of Health, Tehran University of Medical Sciences, Tehran, Iran 1 Department of Microbiology, Faculty of Basic Sciences, Qom Branch, Islamic Azad University, Qom, Iran 2 MARS Bioinformatics Institute, Tehran, Iran 3 Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

Abstract Aims: Staphylococcus aureus is an important nosocomial infectious agent. The prevalence of antibiotic resistance complicates the treatment of staphylococcal infections. Therefore, the development of an effective vaccine against Staphylococcus aureus is important. saCOL2291 and saCOL2581 are potential candidate genes for vaccine development and their sequences are available in Gene Bank. This study was carried out with the aim of determining the mutations in different isolates and confirming the protein of these genes. Materials & Methods: Genomic DNA was extracted from 30 clinical Staphylococcus aureus isolates, randomly obtained from urine, ulcer, mucosa and CSF samples of the inpatients of different Iranian hospitals. The genomic DNA was extracted after culture and the genes were PCR-amplified. The amplicons were sequenced and aligned with Puls4 software. Then, the nucleotide sequences were changed into amino-acid sequences by Gene runner software and the protein sequences were aligned and analyzed. Results: Nucleotide polymorphisms resulted in amino-acid sequence changes. These polymorphisms were located in the N-terminal domains, while the C-terminal domains of these genes were conserved. Conclusion: Both genes are appropriate candidates for vaccine due to their presence in most Staphylococcus aureus species and the existence of conserved C-terminal domain. Keywords: Vaccine, saCOL, Staphylococcus aureus

.‫ فزستبده شوند‬[email protected] ‫ تمبم درخواستهب ببید به نشبنی‬.‫ محمدرضب پورمند‬:‫نویسنده مسئول‬ 00/3/11 :‫ پذیزش مقبله‬00/1/22 :‫دریبفت مقبله‬

PVL (egc)

C

Sca Sca

saCOL2581

ScaD

MRSA

ScaC

saCOL2291

mec

saCOL2291 saCOL2581

CSF DNase ABC DNA LB DNA

SA2006 FnbpB

FnbpA

A

SA1472 SA2291

saCOL2291 DNA

saCOL2581 DNA Pfu

A

(NC_2951.2) PCR (NC_002951.2) saCOL2581 saCOL2291 NCBI PCR

COL

TSST-1

saCOL2291 saCOL2581

saCOL2581 saCOL2291 PCR

´

´

GCGCGCCATATGTCTGAGCAAGATAACTACGGTT

saCOL2291

GCGCGCCTCGAGGTGAATGAAGTTATAACCAGCAG GCGCGCCATATGGCAGGACTTGCCACTATCGC

saCOL2581

GCGCGCCTCGAGATGAATGAAATTATATGAACCTGC

ScaC

ScaD BLAST

COL Puls4

saCOL2291 saCOL2291

bp ssaA2

COL

N

C5

Gene runner

ssaA2

2

1

M

1000bp 500bp

N C bp ssaA1 COL

saCOL2581 saCOL2581

ssaA1 C5

saCOL2291

PCR

SM0311

M

saCOL2581

saCOL2581 saCOL2291

saCOL2581

N C

saCOL2291

saCOL2581 saCOL2291

PCR

saCOL2291 saCOL2581 COL saCOL2291 saCOL2581 C saCOL2581 Kowsar Medical Journal

Vol. 16, No. 3, Fall 2011

saCOL2291

(PBP2a)

saCOL2581 saCOL2581 TATAAC (181-187) [6] GCTACA (178-184) [6]

CAT(40-42) [3]

A(331)  T [5] G(112)  A [6]

202bp [1]

30bp [1] T(514)  A [5]

T(165)  C [9] T(267)  C [6] C(342)  T [6] C(390)  T [30] T(432)  C [30] C(492)  T [10] C(498)  T [6]

saCOL2291

saCOL2581

saCOL2581 saCOL2581 Tyr+Asn#(61-62) [6] Ala+Tyr^(60-61) [6]

Asn(55)  Asn [9] Asn(89)  Asn [6] Pro(130)  Ser [30] Ala(114)  Val [6] Gly(166)  Gly [6] Arg(164)  Trp [10] Ala(144)  Val [30]

Asn(111)  Tyr [5]

His* [3]

Asp(38)  Asn [6] -----

---Ala(172)  Val [5]

^

saCOL

ScaC

ScaD

#

*

IsdB Sca (Sca) C

IsdB IsdA SdrE SdrD IsdB ScaD ScaC

saCOL2581 saCOL2291 COL N

COL saCOL2291 saCOL2291

COL DNA Pfu saCOL2581

saCOL2581

saCOL2291 saCOL2581

Huber LA, et al. Identification of vaccine candidate antigens of Staphylococcus aureus by serological proteome analysis. Proteomics. 2002;2(5):580-90. 9- Foster TJ. Potential for vaccination against infections caused by Staphylococcus aureus vaccine. Vaccine. 1991;9(4):221-7. 10- Hall AE, Domanski PJ, Patel PR, Vernachio JH, Syribeys PJ, Gorovits EL, et al. Characterization of a protective monoclonal antibody recognizing Staphylococcus aureus MSCRAMM protein clumping factor A. Infect Immun. 2003;71(12):6864-70. 11- Hu DL, Omoe K, Sasaki S, Sashinami H, Sakuraba H, Yokomizo Y, et al. Vaccination with nontoxic mutant toxic shock syndrome toxin 1 protects against Staphylococcus aureus infection. J Infect Dis. 2003;188(5):743-52. 12- Dumitrescu O, Tristan A, Meugnier H, Bes M, Gouy M, Etienne J, et al. Polymorphism of the Staphylococcus aureus Panton-valentine leukocidin genes and its possible link with the fitness of community-associated methicilin-resistant S. aureus. Infect Dis. 2008;198(5):792-4. 13- Blaiotta G, Fusco V, VonEiff C, Villani F, Becker K. Biotyping of entritoxigenic Staphylococcus aureus by entrotoxin gene cluster (egc) polymorphism. Appl Environ Mcrobial. 2006;72(9):6117-23. 14- Kim YK, Kim JS, Kim HS, Song W, Cho HC, Lee KM. Molecular typing of Staphylococcus aureus isolated from blood on the basis of coagulase gene polymorphism and toxin genes. Korean J Lab Med. 2008;28(4):286-92. 15- Silva ER, Boechat JU, Silva N. Coagulase gene polymorphism of Staphylococcus aureus isolated from goat mastitis in Brazilaien dairy herds. Lett Appl Microbial. 2006;42(1):30-4. 16- Shinefield H, Black S, Fattom A, Horwith G, Rasgon S, Ordonez J, et al. Use of a Staphylococcus aureus conjugate vaccine in patients receiving hemodialysis. N Engl J Med. 2002;346(7):491-6. 17- Senna JP, Roth DM. Protective immune response against methicillin-resistant Staphylococcus aureus in a murine model using a DNA vaccine approach. Vaccine. 2003;21(1920):2661-6. 18- Nelly AK, Desmond JC, Susan S, James C, Leslie DC, Tessie M, et al. A novel Staphylococcus aureus vaccine: Iron surface determinant B induces rapid antibody responses in Rhesus macaques and specific increased survival in a murine S. aureus sepsis model. Infec Immun. 2006;74(4):2215-23. 19- Yukiko K. Stranger J, Taeok B, Olaf S. Vaccine assembly from surface proteins of Staphylococcus aureus. Emil C, editor. New York: Rockefeller University Publication; 2006. 20- Dryla A, Prustomersky S, Gelbmann D, Hanner M, Beetinger E, Kocsis B. Comparison of antibody repertoires against Staphylococcus aureus in healthy individuals and in acutely infected patients. Clin Diagn Lab Immun. 2005;12(3):387-39. 21- Gill SR, Fouts DE, Archer GL, Mongodin EF, DeBoy RT, Ravel J, et al. Insights on evolution of virulence and resistance from the complete genome analysis of an early methicillinresistant Staphylococcus aureus strain and a biofilm-producing methicillin-resistant Staphylococcus epidermidis strain. J Bacteriol. 2005;187(7):2426-38. 22- Menbari SH, Pourmand MR, Shirazi MH, Mardani N. Cloning and sequencing of Sacol a novel gene from Staphylococcus aureus. Iran Med Microbial J. 2008;2(1):9-14.

Kowsar Medical Journal

Vol. 16, No. 3, Fall 2011

saCOL2581 saCOL2291

C

C

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