Molecular Study of Panton-Valentine Leukocidin Genes among Staphylococcus aureus Clinical Isolates in Khartoum State, Sudan

Najem Aldin M. Osman^1, , Intisar E. Alrayah², Yassir Mahgoub Mohamed³, Ali M El-Eragi⁴, Muataz M. Eldirdery³ and Mohamed Ahmed Salih⁵

¹Department of Biotechnology, Faculty of Science and Technology, Omdurman Islamic University, Sudan

²College of Applied Medical Science, Shaqra University, KSA

³Department of Microbiology, Tropical Medicine Research Institute, National Center for Research, Sudan

⁴Department of Microbiology, Faculty of Medicine, King Khalid University, KSA

⁵Department of Biotechnology, Biotechnology Park, Africa City of Technology, Sudan

Cite this paper:
Najem Aldin M. Osman, Intisar E. Alrayah, Yassir Mahgoub Mohamed, Ali M El-Eragi, Muataz M. Eldirdery and Mohamed Ahmed Salih. Molecular Study of Panton-Valentine Leukocidin Genes among Staphylococcus aureus Clinical Isolates in Khartoum State, Sudan. American Journal of Microbiological Research. 2015; 3(3):107-111. doi: 10.12691/ajmr-3-3-2

Abstract

Staphylococcus aureus strains carrying Panton- Valentine Leukocidin genes (PVL) are an emerging threat worldwide, causing variety of infections even in healthy individuals. Intensive efforts through the last years have been carried out towards the detection and analysis of PVL genes. The prevalence and characterization of such genes has not been done in Sudan. In this study we investigated the prevalence and the molecular characteristics of PVL genes among S. aureus clinical isolates, comparing their PVL allelic variant with that of PVL positive strains from different countries. Standard microbiological procedures were used for the identification of isolates, polymerase chain reaction for determination of PVL genes and standard sequencing for mapping of lukS/F-PV genes. In Silico tools were used for sequence analysis. Among 210 S. aureus isolates, PVL genes were detected in 122 (58%). Sequence analysis for lukS/F-PV genes from 12 representative isolates detected a new point mutation in lukS PV region. Collectively, our findings showed a high frequency of PVL genes among S. aureus isolates and revealed a novel nonsynonymous mutation. Phylogenetic analysis revealed that Sudanese isolates were closely related to each other.

Keywords:
PVL S. aureus Sudan

This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

Figures

References:

[1]	Holmes A, Ganner M, McGuane S, Pitt TL, Cookson BD, Kearns AM: Staphylococcus aureus isolates carrying Panton-Valentine leucocidin genes in England and Wales: frequency, characterization, and association with clinical disease. Journal of clinical microbiology 2005, 43(5):2384-2390.
[2]	Prevost G, Mourey L, Colin DA, Menestrina G: Staphylococcal pore-forming toxins. Current topics in microbiology and immunology 2001, 257:53-83.
[3]	Ward PD, Turner WH: Identification of staphylococcal Panton-Valentine leukocidin as a potent dermonecrotic toxin. Infection and immunity 1980, 28(2):393-397.
[4]	Prevost G, Cribier B, Couppie P, Petiau P, Supersac G, Finck-Barbancon V, Monteil H, Piemont Y: Panton-Valentine leucocidin and gamma-hemolysin from Staphylococcus aureus ATCC 49775 are encoded by distinct genetic loci and have different biological activities. Infection and immunity 1995, 63(10):4121-4129.
[5]	Lina G, Piemont Y, Godail-Gamot F, Bes M, Peter MO, Gauduchon V, Vandenesch F, Etienne J: Involvement of Panton-Valentine leukocidin-producing Staphylococcus aureus in primary skin infections and pneumonia. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America 1999, 29(5):1128-1132.
[6]	Diep BA, Sensabaugh GF, Somboonna N, Carleton HA, Perdreau-Remington F: Widespread skin and soft-tissue infections due to two methicillin-resistant Staphylococcus aureus strains harboring the genes for Panton-Valentine leucocidin. Journal of clinical microbiology 2004, 42(5):2080-2084.
[7]	McClure J, Conly JM, Lau V, Elsayed S, Louie T, Hutchins W, and Zhang K: Novel multiplex PCR assay for detection of Staphylococcus virulence marker Panton-Valentine Leukocidin genes and simultaneous discrimination of Methicillin-susceptible from resistant staphylococci. Novel multiplex PCR assay for detection of Staphylococcus virulence marker Panton-Valentine Leukocidin genes and simultaneous discrimination of Methicillin-susceptible from resistant staphylococci 2006, 44(3):1141-1144.
[8]	Naas T, Fortineau N, Spicq C, Robert J, Jarlier V, and Nordman P: Three years survey of community acquired methicillin-resistant staphylococcus aureus producing Panton-Valentine Leukocidin in French university hospital. . J Hosp Infect 2005 61:321-329.
[9]	Steward CD, Raney PM, Morrell AK, Williams PP, McDougal LK, Jevitt L, McGowan JE, Jr., Tenover FC: Testing for induction of clindamycin resistance in erythromycin-resistant isolates of Staphylococcus aureus. Journal of clinical microbiology 2005, 43(4):1716-1721.
[10]	Etienne J: Panton-Valentine leukocidin: a marker of severity for Staphylococcus aureus infection? Clinical infectious diseases : an official publication of the Infectious Diseases Society of America 2005, 41(5):591-593.
[11]	O'Hara FP, Guex N, Word JM, Miller LA, Becker JA, Walsh SL, Scangarella NE, West JM, Shawar RM, Amrine-Madsen H: A geographic variant of the Staphylococcus aureus Panton-Valentine leukocidin toxin and the origin of community-associated methicillin-resistant S. aureus USA300. The Journal of infectious diseases 2008, 197(2):187-194.
[12]	Otokunefor K, Sloan T, Kearns AM, James R: Molecular characterization and panton-valentine leucocidin typing of community-acquired methicillin-sensitive Staphylococcus aureus clinical isolates. Journal of clinical microbiology 2012, 50(9):3069-3072.
[13]	Nakagawa S, Taneike I, Mimura D, Iwakura N, Nakayama T, Emura T, Kitatsuji M, Fujimoto A, Yamamoto T: Gene sequences and specific detection for Panton-Valentine leukocidin. Biochemical and biophysical research communications 2005, 328(4):995-1002.
[14]	Berglund C, Prevost G, Laventie BJ, Keller D, Soderquist B: The genes for Panton Valentine leukocidin (PVL) are conserved in diverse lines of methicillin-resistant and methicillin-susceptible Staphylococcus aureus. Microbes and infection / Institut Pasteur 2008, 10(8):878-884.
[15]	Dumitrescu O, Tristan A, Meugnier H, Bes M, Gouy M, Etienne J, Lina G, Vandenesch F: Polymorphism of the Staphylococcus aureus Panton-Valentine leukocidin genes and its possible link with the fitness of community-associated methicillin-resistant S. aureus. The Journal of infectious diseases 2008, 198(5):792-794.
[16]	Cowan ST, Steel KJ, Barrow GI, Feltham RKA: Cowan and Steel's manual for the identification of medical bacteria, 3rd edn. Cambridge ; New York: Cambridge University Press; 1993.
[17]	Collee JG, Duguid JP, Fraser AG, and Marmion BP: Mackie & MacCarteny practical medical microbiology., 13 edn. London: Churchill Livingstone; 1989.
[18]	Atschul S, Madden TL, al SAe: Gapped BLAST and PSI-BLAST. A new generation of protein database search programmes. NucleicAcid Res 1997, 25:3389-3402.
[19]	Hall T: BioEdit: a user-friendly biologicalsequence alignment editor and analysis programfor Windows 95/98/NT. Nucl Acids Symp Ser 1999, 41:95-98.
[20]	Besemer J, Lomsadze A, Borodovsky M: GeneMarkS: a self-training method for prediction of gene starts in microbial genomes. Implications for finding sequence motifs in regulatory regions. Nucleic acids research 2001, 29(12):2607-2618.
[21]	Dereeper A, Guignon V, Blanc G, Audic S, Buffet S, Chevenet F, Dufayard JF, Guindon S, Lefort V, Lescot M et al: Phylogeny.fr: robust phylogenetic analysis for the non-specialist. Nucleic acids research 2008, 36(Web Server issue):W465-469.
[22]	Tamura K, Stecher G, Peterson D, Filipski A, Kumar S: MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular biology and evolution 2013, 30(12):2725-2729.
[23]	Ramdani-Bouguessa N, Bes M, Meugnier H, Forey F, Reverdy ME, Lina G, Vandenesch F, Tazir M, Etienne J: Detection of methicillin-resistant Staphylococcus aureus strains resistant to multiple antibiotics and carrying the Panton-Valentine leukocidin genes in an Algiers hospital. Antimicrobial agents and chemotherapy 2006, 50(3):1083-1085.
[24]	Steward CD RP, Morrell AK, Williams PP, McDougal LK, and Jevitt, L.: Testing for Induction of Clindamycin Resistance in Erythromycin-Resistant Isolates of Staphylococcus aureus. Journal of clinical microbiology 2005, 43:1716-1721.
[25]	Bocchini CE, Hulten KG, Mason EO, Jr., Gonzalez BE, Hammerman WA, Kaplan SL: Panton-Valentine leukocidin genes are associated with enhanced inflammatory response and local disease in acute hematogenous Staphylococcus aureus osteomyelitis in children. Pediatrics 2006, 117(2):433-440.
[26]	Labandeira-Rey M, Couzon F, Boisset S, Brown EL, Bes M, Benito Y, Barbu EM, Vazquez V, Hook M, Etienne J et al: Staphylococcus aureus Panton-Valentine leukocidin causes necrotizing pneumonia. Science 2007, 315(5815):1130-1133.
[27]	Enany S, Yaoita E, Yoshida Y, Enany M, Yamamoto T: Molecular characterization of Panton-Valentine leukocidin-positive community-acquired methicillin-resistant Staphylococcus aureus isolates in Egypt. Microbiological research 2010, 165(2):152-162.