This document presents a study on the detection of methicillin-resistant Staphylococcus aureus (MRSA) isolated from different clinical samples at a hospital laboratory. The study aims to isolate S. aureus from samples using culture and biochemical tests, perform antibiotic susceptibility testing, and evaluate antimicrobial resistance patterns to determine the presence of MRSA. The methodology describes sample collection and testing procedures. Expected outcomes anticipate finding MRSA in 5% of isolated S. aureus samples. A timeline and budget are provided to complete laboratory work, analysis, and thesis submission within 3 months with a budget of 10,000 Rupees.

1. DETECTION OF METHICILLIN
RESISTANT STAPHYLOCOCCUS AUREUS I
SOLATED FROM DIFFERENT CLINICAL
SAMPLES OF HOSPITAL LAB
Presented by
• Shambhu Rauniyar
• Geetanjali Yadav
• Rakesh Thakur
• Ramesh Paudel
• Bishal babu Katel
Supervisor: Mr. Janak Raj Dhungana

2. INTRODUCTION
• What is MRSA?
S. aureus is gram positive cocci arranged in grape like
clusters, halotolerant and produces a golden yellow
pigment on agar media.
Methicillin-resistant Staphylococcus aureus (MRSA)
is by definition a strain of Staphylococcus aureus that
is resistant to a large group of antibiotics called the β-
lactams, which include the penicillin and the
cephalosporins.
MRSA were first reported in the early 1960's and
now regarded as major hospital acquired pathogen
worldwide. The term methicillin resistant is used to

3. RATIONALE
• Why study on MRSA is important?
 MRSA can cause life-threatening skin infections as
well as infections in bones, joints, surgical wounds,
heart valves and lungs.
MRSA was isolated at the rate 75.5% from clinical
samples in a study conducted by Rijal et al. in Pokhara
Valley (Rijal et al. 2008).
As of 2013 US CDC threat report, over 80,000 severe
MRSA infections occur every year and among those
11,000 do not survive.
This make a great concern for us as a researchers
regarding public heath.

4. OBJECTIVES
• General objectives:
The purpose of this study was to assess the extent of
Methicillin Resistant Staphylococcus aureus from
different clinical samples of patients visiting the
hospital lab.
• Specific objectives:
To isolate S. aureus from the samples.
To confirm presence of S. aureus via biochemical test
To perform antimicrobial susceptibility testing by
modified Kirby-Bauer disk diffusion method.
To evaluate the AST pattern of the isolates to a set of
antibiotics and determination of methicillin resistance.

5. METHODOLOGY
Clinical Samples
Culture on
BA,MA
Identification of isolates
by gram staining and biochemical test
Inoculation on MSA
(selective media for S. aureus)
+ve -ve
Catalase: +ve
Coagulase: +v
Oxidase: -ve
discard
Clusters
i.e. S. aureus
Antibiotic Susceptibility Test
Identification & reporting of result
Screening & detection of MRSA

6. EXPECTED OUTCOMES
There may be presence of at least 20% of S. aureus
among which 5% of S. aureus may be detected as
methicillin resistant.
Hence few no. of MRSA may be found due to the
resistant property developed in S. aureus against
different derivatives of β lactum antibiotics.
The result is expected to be positive and significant.

7. GANTT CHART
S.
N
HEADINGS 1st
month
2nd
month
3rd
month
1. Literature Review
2. Conceptual
framework
3. Experiment design
4. Sample collection
5. Lab work
6. Data analysis &
interpretation
7. Submission of
thesis

8. BUDGET
• The estimated budget for this project is Rs.10,000/-
for each. The whole allocated amount will be used in:
Lab practice: 5*5,000 = 25,000/-
Transportation: 3,000/-
Printing of this project: 500/-
Miscellaneous expenses: 2000/-
Lab expenditure: 20,000/-
oTest tubes
oChemical reagents
oDifferent agar
oDifferent media
oAntibiotic disk
oOthers

9. REFERENCES
• Chowdhury T, MahmudA, BaruaA, Khalil M, Chowdhury R,Ahamed F
and Dhar K( 2016). Bacterial contaminations on hand touch surfaces of
public busses in Chittagong City, Bangladesh. J Env SciToxic Food
Tech 10: 48-55.
• CS VK, Prasad S, Patil S, ReddyA, JhaA, KG RK, VLJ and KG B
(2017). Public transport: alarge scale fomite of methicillin-resistant
Staphylococcus aureus. Int J Res Med Sci 6(1): 172-176.
• Carleton H, Diep B, Charlebois E and Sensabaugh G(2004).
Community-adapted methicillin-resistant Staphylococcus aureus
(MRSA): population dynamics of an expanding community reservoir of
MRSA. JInfect Dis 190(10): 1730-1738.
• IwaoY, Shizuka Y,Takano T, Higuchi W,NishiyamaAandYamamoto T
(2012). Isolation and molecular characterization of methicillin-resistant
Staphylococcus aureus from public transport. Microbiol Immunol 56(1):
76-82

10. THANK YOU