Study of genetic variation in Staphylococcus aureus strains isolated from clinical specimens by RAPD-PCR Technique
Subject Areas : microbiologyBaroomand Morvaridi 1 , Shahram Nakhjavan 2 , Shahram Nanekarani 3 , Reza Yari 4
1 - MSc. Student, Department of Biology, Borujerd Branch, Islamic Azad University, Borujerd, Iran.
2 - Assistant Professor, Department of Agronomy and Plant Breeding, Borujerd
Branch, Islamic Azad University, Borujerd, Iran.
3 - Assistant Professor, Department of Animal Science,Borujerd Branch, Islamic Azad University, Borujerd, Iran.
4 - Assistant Professor. Department of Biology, Borujerd Branch, Islamic Azad University, Borujerd,
Iran
Keywords: Genetic variation, RAPD-PCR, Methicillin Resistant-Staphylococcus aureus, Qom City,
Abstract :
Introduction: Methicillin Resistant-Staphylococcus aureus (MRSA) has been introduced as important factor in Hospital-Acquired Infections (HAI). Diversity in Staphylococcus strains can affect the response to treatment and identifying of strains can be effective in the selection of antibiotics. The aim of this study was to determine the genetic diversity of MRSA isolates with random primers. Material and Methods: This cross-sectional descriptive study was performed in 2015 on 50 MRSA isolates of clinical skin samples. Genomes were extracted by Boiling method. RAPD-PCR method was performed by 16 random primers. To investigate the genetic similarity, matrix 1/0, NTSYS and MVSP software's were used. One-dimensional clustering and ordinations were conducted. Results: The highest and lowest produced bonds related to M4 and M9 primers respectively. The greatest mean produced bands for each isolate/primer up to 8.1 relates to M4 primer. The most polymorphic bands, 36 bands belonged to M5 primer. The heaviest band, 3.6 Kb produced by M2 primer and the lightest band, 100 bp produced by M12 primer. A total of 16 primers, 583 bands formed that were 412 (70.91%) polymorphic bands and 171 (29.09%) specific bands. RAPD-PCR divided isolates into two clusters. Conclusion: This study showed that these isolates are very heterogeneous genetically. RAPD-PCR is important in rapid detection of an epidemic outbreak, tracing the origin of the infection and the subsequent managing of infection control. Therefore, the present study tries to develop this approach and apply it in health management by providing information about the genetic diversity of MRSA.
Nowroozi J, Goudarzi G, Pakzad P, Razavipour R. Isolation and Detection of Staphylococcus aureus Enterotoxins A-E and TSST-1 Genes from Different Sources by PCR Method. J Qom Uni Med Sci. 2012; 6(3): 78-85. [In Persian]
Gadyari F, Sattari M, Boroumand M, Yaghoubi R, Sepehriseresht S, Purgholi L. Detection of Staphylococcus aureus Entrotoxins A to D in clinical strains isolated
from burned patients of Tehran Motahari Hospital. J Iran Med Sci. 2011; 5(5): 20-27.
[In Persian]
Kurlenda J, Grinholc M, Jasek K, Wegrzyn G. RAPD typing of methicillin-resistant Staphylococcus aureus: A 7-year experience in a Polish hospital. Diagn Med Tecnol. 2007; 13(6): 13-18.
Pereira MSV, Leal NC, Leal TCA, Sobreira M, de Almeida AMP, Siqueira-Junior JP, Campos-Takaki GM. Typing of human and bovine Staphylococcus aureus by RAPD-PCR and ribotyping-PCR. Lett Appl Microbiol. 2002; 35: 32–36.
Naffa RG, Bdour SM, Migdadi HM, Shehabi AA. Enterotoxicity and genetic variation among clinical Staphylococcus aureus isolates in Jordan. J Med Microbiol. 2006; 55(2): 183–187.
Yari R, Mehrabi MR, Gorbanpoor AmirKiasari N. Detection of Methicillin-resistant Staphylococcus aureus Strains by Comparing Oxacillin and Cefoxitin Disk Diffusion and PCR for mecA Gene. Appl Biol. 2016; 6(3): 40-50. [In Persian]
Andrasevic AT, Power EG, Anthony RM, et al. Failure of bacteriophage typing to detect an inter-hospital outbreak of methicillin-resistant Staphylococcus aureus (MRSA) in Zagreb subsequently identifi ed by random amplification of polymorphic DNA (RAPD) and Pulsed-Field Gel Electrophoresis (PFGE). Clin Microbiol Infect. 1999; 10: 634–642.
Blanc DS, Francioli P, Hauser PM. Poor value of Pulsed-Field Gel Electrophoresis to investigate long-term scale epidemiology of methicillin resistant Staphylococcus aureus. Infect Genet Evol, 2002; 2(2): 145–148.
Telecco S, Barbarini D, Carretto E, Comincini S, Emmi V, Marone P. Typing of methicillin-resistant Staphylococcus aureus (MRSA) strains from an intensive care unit by Random Amplified Polymorphic DNA (RAPD). New Microbiol, 1999; 22(4): 323–329.
Al-Thawadi SI, Kessie G, Dela Cruz D, Al-Ahdal MN. A comparative study on the application of 3 molecular methods in epidemiological typing of bacterial isolates using MRSA as a prototype. Saudi Med J, 2003; 24(12): 1317–1324.
Reinoso E, Bettera S, Frigerio C, Direnzo M, Calzolari A, Bogni C. RAPD-PCR analysis of Staphylococcus aureus strains isolated from bovine and human hosts. Microbiol Res. 2004; 1(59): 245–255.
Idil N, Bilkay IS. Application of RAPD-PCR for determining the clonality of Methicillin Resistant Staphylococcus aureus isolated from different hospitals. Braz. Arch. Biol. Technol. 2014; 57(4): 548-553.
Deplano A, Vaneechoutte M, Verschraegen G, Struelens MJ. Typing of Staphylococcus strains by PCR analysis of inter Is256 spacer length polymerase. J Clin Microbiol. 1997; 35(10): 2580-2587.
Hakimi Alni R, Mohammadzadeh A, Mahmmodi P, Alikhani Y. RAPD-PCR analysis of Staphylococcus aureus strains isolated from different sources. Comp Clin Pathol. 2017; 26(4): 823-830.
Byun DE, Kim SH, Shin JH, Suh SP, Ryang DW. Molecular epidemiological analysis of Staphylococcus aureus isolated from clinical specimens. J Korean Med Sci. 1997; 12(3): 190-198.
_||_Nowroozi J, Goudarzi G, Pakzad P, Razavipour R. Isolation and Detection of Staphylococcus aureus Enterotoxins A-E and TSST-1 Genes from Different Sources by PCR Method. J Qom Uni Med Sci. 2012; 6(3): 78-85. [In Persian]
Gadyari F, Sattari M, Boroumand M, Yaghoubi R, Sepehriseresht S, Purgholi L. Detection of Staphylococcus aureus Entrotoxins A to D in clinical strains isolated
from burned patients of Tehran Motahari Hospital. J Iran Med Sci. 2011; 5(5): 20-27.
[In Persian]
Kurlenda J, Grinholc M, Jasek K, Wegrzyn G. RAPD typing of methicillin-resistant Staphylococcus aureus: A 7-year experience in a Polish hospital. Diagn Med Tecnol. 2007; 13(6): 13-18.
Pereira MSV, Leal NC, Leal TCA, Sobreira M, de Almeida AMP, Siqueira-Junior JP, Campos-Takaki GM. Typing of human and bovine Staphylococcus aureus by RAPD-PCR and ribotyping-PCR. Lett Appl Microbiol. 2002; 35: 32–36.
Naffa RG, Bdour SM, Migdadi HM, Shehabi AA. Enterotoxicity and genetic variation among clinical Staphylococcus aureus isolates in Jordan. J Med Microbiol. 2006; 55(2): 183–187.
Yari R, Mehrabi MR, Gorbanpoor AmirKiasari N. Detection of Methicillin-resistant Staphylococcus aureus Strains by Comparing Oxacillin and Cefoxitin Disk Diffusion and PCR for mecA Gene. Appl Biol. 2016; 6(3): 40-50. [In Persian]
Andrasevic AT, Power EG, Anthony RM, et al. Failure of bacteriophage typing to detect an inter-hospital outbreak of methicillin-resistant Staphylococcus aureus (MRSA) in Zagreb subsequently identifi ed by random amplification of polymorphic DNA (RAPD) and Pulsed-Field Gel Electrophoresis (PFGE). Clin Microbiol Infect. 1999; 10: 634–642.
Blanc DS, Francioli P, Hauser PM. Poor value of Pulsed-Field Gel Electrophoresis to investigate long-term scale epidemiology of methicillin resistant Staphylococcus aureus. Infect Genet Evol, 2002; 2(2): 145–148.
Telecco S, Barbarini D, Carretto E, Comincini S, Emmi V, Marone P. Typing of methicillin-resistant Staphylococcus aureus (MRSA) strains from an intensive care unit by Random Amplified Polymorphic DNA (RAPD). New Microbiol, 1999; 22(4): 323–329.
Al-Thawadi SI, Kessie G, Dela Cruz D, Al-Ahdal MN. A comparative study on the application of 3 molecular methods in epidemiological typing of bacterial isolates using MRSA as a prototype. Saudi Med J, 2003; 24(12): 1317–1324.
Reinoso E, Bettera S, Frigerio C, Direnzo M, Calzolari A, Bogni C. RAPD-PCR analysis of Staphylococcus aureus strains isolated from bovine and human hosts. Microbiol Res. 2004; 1(59): 245–255.
Idil N, Bilkay IS. Application of RAPD-PCR for determining the clonality of Methicillin Resistant Staphylococcus aureus isolated from different hospitals. Braz. Arch. Biol. Technol. 2014; 57(4): 548-553.
Deplano A, Vaneechoutte M, Verschraegen G, Struelens MJ. Typing of Staphylococcus strains by PCR analysis of inter Is256 spacer length polymerase. J Clin Microbiol. 1997; 35(10): 2580-2587.
Hakimi Alni R, Mohammadzadeh A, Mahmmodi P, Alikhani Y. RAPD-PCR analysis of Staphylococcus aureus strains isolated from different sources. Comp Clin Pathol. 2017; 26(4): 823-830.
Byun DE, Kim SH, Shin JH, Suh SP, Ryang DW. Molecular epidemiological analysis of Staphylococcus aureus isolated from clinical specimens. J Korean Med Sci. 1997; 12(3): 190-198.