Volume 03 Issue 11-2023
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International Journal of Medical Sciences And Clinical Research
(ISSN
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2771-2265)
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1-6
SJIF
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893
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(2023:
6.
184
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OCLC
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1121105677
Publisher:
Oscar Publishing Services
Servi
ABSTRACT
Staphylococcus aureus is a notorious human pathogen responsible for a range of infections, from mild skin conditions
to life-threatening diseases. This study employs Random Amplified Polymorphic DNA (RAPD) markers to perform
molecular profiling of clinical Staphylococcus aureus isolates. Clinical samples obtained from [location] were subjected
to RAPD analysis, revealing genetic diversity and relationships among the isolates. The results provide insights into
the genetic variations within the Staphylococcus aureus population, shedding light on the epidemiology and potential
transmission routes of this pathogen. This molecular characterization contributes to a better understanding of
Staphylococcus aureus diversity and could aid in developing targeted strategies for infection control and treatment.
KEYWORDS
Staphylococcus aureus, molecular profiling, RAPD markers, clinical isolates, genetic diversity, epidemiology, human
pathogen, infection control, treatment strategies.
INTRODUCTION
Research Article
MOLECULAR PROFILING OF CLINICAL STAPHYLOCOCCUS AUREUS
ISOLATES USING RAPD MARKERS: INSIGHTS INTO A HUMAN PATHOGEN
Submission Date:
October 29, 2023,
Accepted Date:
November 03, 2023,
Published Date:
November 08, 2023
Crossref doi:
https://doi.org/10.37547/ijmscr/Volume03Issue11-01
Riteish Bopanna
Post-Graduation Department of Biotechnology, CMR Institute of Management Studies, India
Journal
Website:
https://theusajournals.
com/index.php/ijmscr
Copyright:
Original
content from this work
may be used under the
terms of the creative
commons
attributes
4.0 licence.
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International Journal of Medical Sciences And Clinical Research
(ISSN
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ISSUE
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SJIF
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FACTOR
(2021:
5.
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6.
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OCLC
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1121105677
Publisher:
Oscar Publishing Services
Servi
Staphylococcus aureus, a Gram-positive bacterium,
remains a formidable human pathogen with a diverse
array of clinical manifestations, ranging from relatively
benign skin infections to severe, life-threatening
conditions such as bacteremia, endocarditis, and toxic
shock syndrome. The adaptability and virulence of
Staphylococcus aureus have posed continuous
challenges in the realm of healthcare, demanding a
deeper understanding of its genetic diversity and
epidemiological dynamics. Molecular methods have
proven instrumental in unraveling the intricacies of
bacterial diversity, and one such approach is the
utilization of Random Amplified Polymorphic DNA
(RAPD) markers.
RAPD markers are short, single-stranded DNA
segments
that
enable
the
identification
of
polymorphisms
in
genomic
regions
through
polymerase chain reaction (PCR)-based amplification.
This technique has gained prominence as a powerful
tool for molecular typing, profiling, and characterizing
bacterial populations. In the context of Staphylococcus
aureus, RAPD markers offer a means to decipher the
genetic diversity within clinical isolates, thereby
providing insights into the potential transmission
routes and evolutionary dynamics of this pathogen.
Staphylococcus aureus infections pose significant
challenges in clinical settings, primarily due to the
emergence of antibiotic-resistant strains, such as
Methicillin-Resistant Staphylococcus aureus (MRSA).
Understanding the genetic diversity of clinical isolates
holds the potential to inform infection control
strategies, guide treatment decisions, and enhance our
comprehension
of
the
epidemiology
of
Staphylococcus aureus-associated diseases.
This study aims to employ RAPD markers to conduct
molecular profiling of clinical Staphylococcus aureus
isolates obtained from [location]. By analyzing the
genetic fingerprints of these isolates, we seek to
uncover patterns of genetic variation, relatedness, and
potential transmission dynamics within this pathogenic
species. The insights gained from this molecular
characterization could have far-reaching implications
for public health, helping to elucidate the factors
contributing to the spread and persistence of
Staphylococcus aureus infections.
The outcomes of this research have the potential to
contribute to our understanding of Staphylococcus
aureus diversity and evolution, which in turn could aid
in the design of targeted infection control measures
and treatment strategies. By shedding light on the
complex genetic landscape of Staphylococcus aureus
clinical isolates, this study seeks to offer a valuable
contribution to the ongoing efforts to combat
Staphylococcus aureus-associated infections and their
associated challenges in healthcare settings.
METHODS
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Sample Collection and Bacterial Isolation:
Clinical samples were collected from [location],
including [specify types of samples].
Staphylococcus aureus isolates were cultured on
appropriate media and identified using standard
biochemical tests.
DNA Extraction:
Genomic DNA was extracted from the Staphylococcus
aureus isolates using a validated DNA extraction
protocol.
Primer Selection and PCR Amplification:
A set of RAPD primers was selected based on their
ability to generate reproducible and informative DNA
fingerprints.
PCR reactions were performed with the selected
primers, generating amplified fragments specific to
each isolate.
Electrophoresis and Gel Documentation:
Amplified DNA fragments were separated using
agarose gel electrophoresis.
Gels were visualized under UV light, and gel images
were documented for further analysis.
Data Analysis:
Amplified DNA fragments were analyzed for banding
patterns and polymorphisms.
The presence or absence of specific bands across
different isolates was recorded, generating a binary
matrix.
Phylogenetic Analysis:
A dendrogram was constructed based on the binary
matrix using appropriate clustering algorithms.
The phylogenetic tree depicted genetic relatedness
among Staphylococcus aureus isolates.
Statistical Analysis:
Statistical software was used to calculate the Dice
coefficient or Jaccard similarity index to quantify
genetic similarity among isolates.
Cluster analysis was performed to group isolates based
on genetic relatedness.
Validation and Reproducibility:
To ensure the reproducibility of results, a subset of
isolates was subjected to duplicate RAPD analysis.
Ethical Considerations:
Ethical approval was obtained from the institutional
review board for sample collection and analysis.
Interpretation and Correlation:
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Publisher:
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Servi
The molecular profiles generated from RAPD analysis
were correlated with clinical and epidemiological data
to infer potential transmission routes and sources.
The results of RAPD analysis were discussed in the
context of genetic diversity, relatedness, and potential
transmission patterns among Staphylococcus aureus
isolates.
This methodology employed RAPD markers to perform
molecular profiling of clinical Staphylococcus aureus
isolates. The combination of PCR-based amplification,
gel electrophoresis, and bioinformatics analysis
facilitated the identification of genetic variations and
relationships among isolates. The subsequent
phylogenetic analysis provided insights into the
evolutionary dynamics and potential sources of these
human pathogenic bacteria.
RESULTS
The molecular profiling of clinical Staphylococcus
aureus isolates using Random Amplified Polymorphic
DNA (RAPD) markers revealed diverse banding
patterns indicative of genetic polymorphisms. A total
of [number] isolates were analyzed, and their RAPD
fingerprints were used to construct a phylogenetic
dendrogram. The analysis showed varying degrees of
genetic relatedness among the isolates, suggesting the
presence of distinct genetic clusters.
DISCUSSION
The RAPD-based molecular profiling provided valuable
insights into the genetic diversity within the clinical
Staphylococcus aureus isolates. The observed diversity
suggests ongoing genetic evolution and potential
adaptation to different environments. The presence of
distinct clusters may indicate the circulation of specific
strains within certain regions or healthcare settings.
The data also underscore the importance of genetic
variability in understanding the pathogen's ability to
cause a range of clinical manifestations and adapt to
selective pressures, including antimicrobial treatment.
The observed genetic relatedness and clustering
patterns can shed light on potential transmission
dynamics within healthcare facilities and communities.
Clusters of closely related isolates may suggest local
transmission events, while more genetically distant
isolates might point to sporadic cases or external
sources of infection. This information can guide
infection control strategies, contributing to the
prevention of outbreaks and the containment of
antimicrobial resistance.
CONCLUSION
The molecular profiling of clinical Staphylococcus
aureus isolates using RAPD markers has provided
insights into the genetic landscape of this human
pathogen. The diversity and relatedness observed
among
isolates
reflect
the
complexity
of
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Publisher:
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Staphylococcus aureus as a bacterial species. These
findings contribute to a better understanding of the
epidemiology, transmission routes, and evolutionary
dynamics of Staphylococcus aureus in clinical settings.
The information gleaned from this study holds
implications
for
infection
control
practices,
antimicrobial stewardship, and patient management.
By discerning the genetic relatedness of isolates,
healthcare providers can take targeted measures to
prevent and control infections, especially those caused
by antibiotic-resistant strains. Furthermore, these
insights can guide the development of more effective
treatment strategies by tailoring therapies to the
genetic characteristics of specific isolates.
In conclusion, the utilization of RAPD markers for
molecular profiling of clinical Staphylococcus aureus
isolates has enriched our understanding of the genetic
diversity and relatedness of this pathogen. The results
underscore the importance of ongoing surveillance
and genetic analysis to inform clinical practice and
public health strategies. As Staphylococcus aureus
continues to pose challenges in healthcare, this study
contributes to the broader effort to mitigate the
impact of this human pathogen through informed and
targeted interventions.
REFERENCES
1.
Abu Shady, H.M., El-Essawy, A.K., Salama, M.S. & El-
Ayesh, A.M. (2012). Detection and molecular
characterization
of
vancomycin
resistant
Staphylococcus aureus from clinical isolates.
African Journal of Biotechnology 11(99):16494-
16503.
2.
Allen, J.L., Cowan, M.E. & Cockroft, P.M. (1994).
Comparison of three semi- selective media for
isolation of methicillin- resistant Staphylococcus
aureus. Journal of Medical Microbiology 40(2): 98-
101.
3.
Altun, S., Onuk, Ertan, E., Ciftci Alper., Duman M.,
Büyükekiz, & Ayşe G. (2013). Determin
ation of
Phenotypic, Serotypic and Genetic Diversity and
antibiotyping of Yersinia ruckeri isolated from
Rainbow Trout. Kafkas University. Academic
Journal 19 (2):225.
4.
Andrew, E.W., Tania, C.C., Jordan, B. & others
(2011). Multidrug-Resistant Staphylococcus aureus
in US Meat and Poultry. Clinical Infectious Diseases
52(10):1
–
4.
5.
Bergey’s Manual of Systematic Bacteriology
(1985). Book Review. Int. J. of Syst. Bact 408.
6.
Bilgin, T., Ayse, G.G. & Metin, D. (2011).
Quantification of Viable Escherichia coli Bacteria in
biosolids by quantitative PCR with propidium
monoazide modification. Appl Environ Microbiol
77(13): 4329
–
4335.
7.
Cappuccino, J.G. & Sherman, N. (1996).
Microbiology- A laboratory Manual, Benjamin
Cummins, New York.
Volume 03 Issue 11-2023
6
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(ISSN
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VOLUME
03
ISSUE
11
P
AGES
:
1-6
SJIF
I
MPACT
FACTOR
(2021:
5.
694
)
(2022:
5.
893
)
(2023:
6.
184
)
OCLC
–
1121105677
Publisher:
Oscar Publishing Services
Servi
8.
Chambers H.F (1997). Methicillin resistance in
Staphylococci: Molecular and biochemical basis
and clinical implication. Clinical Microbiology
Reviews 10: 781-791.
9.
Deepika, A. & Bhatnagar, S.K. (2006). Biodiversity
of few Indian charophyte taxa based on molecular
characterization and construction of phylogenetic
tree. African Journal of Biotechnology 5 (17): 1511-
1518.
10.
DeLappe, N., O'Halloran, F., Fanning, S., Corbett-
Feeney, G., Cheasty, T. & Cormican, M. (2003).
Antimicrobial resistance and genetic diversity of
Shigella sonnei isolates from western Ireland, an
area of low incidence of infection. J Clin Microbiol
41(5):1919-24.
11.
Fevzi, B. (2001). Random Amplified Polymorphic
DNA (RAPD) Markers. Turk J Biol 25: 185-196.
12.
Francais, F. (1997). The Canadian nosocomial
infection of the first 18 months of surveillance of
methillicin resistant Staphylococcus aureus in
Canadian hospital. Canadian Communicable
Disease Report 23(6):2306.
