World Journal of Zoology 10 (4): 318-322, 2015

ISSN 1817-3098
© IDOSI Publications, 2015
DOI: 10.5829/idosi.wjz.2015.10.4.96146

Antibiotic Susceptibility Patterns of Methicillin Resistant Staphylococcus aureus

at National Institute of Health Sciences, Islamabad, Pakistan
1
Hizbullah, 2Fahad Ali, 1Sulaiman Bahadar, 1Zunaira Shahid, 1Rahimullah,
2
Khalil ur Rahman, 1Afzal Ahmad, 3Abdul Aziz, 4Muhammad Daud, 5Ilyas Khan,
2
Javid Khan, 1Azam Hayat, 1Mujadad-ur-Rahman and 1Muhammad Ayub Khan

1
Department of Microbiology, Abbottabad University of Science and Technology,
Havalian, Abbottabad, Pakistan
2
Department of Genetic, Hazara University Mansehra Dhodial, KhyberPakhtoonkhwa Pakistan
3
Department of Biotechnology and Genetic Engineering,
Kohat University of Science and Technology, Kohat, Pakistan
4
Department of Microbiology, Hazara University Mansehra Dhodial, Khyber Pakhtoonkhwa Pakistan
5
Department of Microbiology, Kohat University of Science and Technology, Kohat, Pakistan

Abstract: Methicillin resistance Staphylococcus aureus is a major public health problem all over the world
especially in progressive countries. As there is an unstoppable change in the prevalence of MRSA, the present
study was designed to investigate the recent trend in the prevalence and antibiotic susceptibility of MRSA.
In current study, 450 clinical samples were assemble at National Institute of Health, (NIH)from January 2015 to
March 2015. Samples were subjected to phenotypic based identification for MRSA and determination of its
antibiotic susceptibility pattern. Out of 450 isolates, 320(71.11%) were confirmed phenotypically as MRSA.
Among these 320 isolates (samples), 190(59.37%) were males and 130(40.62%) were females (samples). The
prevalence of MRSA was found higher in pus 165(51.56%), followed by blood 70(28.17%), urine 59(18.43%) and
sputum 26(8.12%). MRSA positive samples were collected mainly from surgical wards followed by medical
wards, Paediatrics wards and outpatients. All isolates were found sensitive to vancomycin and teicoplanin
while sensitivity to Novobiocin 80.31% clindamycin was (53.12%), chloramphenicol (45%) and ciprofloxacin
was 25%. MRSA is endemic in all units of LRH. It is responsible for increased morbidity and mortality, cost,
hospital stay and poses a great challenge for hospital infection control program.

Key words: MRSA % Nosocomial infections % Antibiotics

INTRODUCTION 1961 [3] MRSA infection first was determined in

Methicillin Resistant Staphylococcus aureus
(MRSA) is the main source of nosocomial infections all
over the world [1]. Soon after its discovery, it was
considered an important pathogen globally in both clinical
practices and communities [2]. Methicillin, which is
semisynthetic penicillin and poorly hydrolyzed by
pencillinase came in clinical practice in the year of 1960.
After a very short usage of this new antibiotic a resistant
strain developed i.e. MRSA emerged unfortunately in
hospitalized patient and there subsequently described as
powerful nosocomial infection [4]. Nosocomial infection
have been started soon when people begin to use
methicillin as antibiotic in 1961. Physicians start giving
methicillin against penicillin resistant staphylococci, but
unfortunately it was no longer effective and soon MRSA
recognized an important human Hospital acquired
infection [5]. MRSA were detected by American physician
in 1970s and later on it was considered endemic in 1990
[6].

Corresponding Author: Sulaiman Bahadar, Department of Microbiology, Abbottabad University of Science and Technology,
Havalian, Abbottabad, Pakistan. E-mail: salupakhtoon@gmail.com.
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 World J. Zool., 10 (4): 318-322, 2015
MRSA came in to being from the methicillin-
susceptible S. aureus (MSSA) by exogenously
acquisition of methicillin resistance gene carried out by a
mobile genetic element known as staphylococcal cassette
chromosome i.e. mec (SCCmec) at 30 end (i.e., 15-bp
SCCmec insertion site, att) or FX of their chromosome
[7, 8]. SCCmec carries a mecA gene that encoding a
penicillin binding protein known as PBP20 which show
resistant toward beta-lactam agents [9].
In industrialized countries the rate of nosocomial
infection is very rare as compare to the developing
countries, it is due to the devastating effort of these
people to overcome the morbidity and mortality rate for
which they follow standard legislative measure. While on
the other hand a rare premature study reported from the
developing countries. According to the World Health
Organization (WHO) in 2001, that nosocomial infection
has the highest in the Eastern Mediterranean and South
East Asia and describe the reason that the hazard use of
antibiotics, overcrowding and unhygienic environment
leads to the increased resistance in the pathogen [10].
Hospital acquired MRSA are frequently multidrug
resistant and poses a constant problem to clinicians and
hospital infection control program. Moreover, the MRSA
situation is getting worse with the passage of time at our
tertiary care level hospital despite so many precautionary
measures. We are a resource challenge society and a
prompt infection control policy is the only way to reduce
this burden [11, 12].
MATERIAL AND METHODS
Study Design and Sampling: The present study was
carried out in NIH a period of three months from January
2015 to march 2015 at the Microbiology Department.
Clinical samples were received from different hospitals e.g
NIH Patients, outsider patients and other hospital patients
were considered in the study. Different types of samples
which include pus, HVS swabs, blood, urine, catheter
tips, CSF, stool and tissue. Samples were collected
aseptically with the help of sterile cotton swab and were
immediately brought to the microbiological laboratory,
department of microbiology, NIH for bacterial isolation
and identification MRSA using standard bacteriological
techniques. (Cruickshark et al.).
Isolation and Identification of S. aureus: Samples was
inoculated on blood agar, XLD (Xylose lysine dextrose)
and mannitol salt agar and the stool samples also
319
were inoculated in peptone water but the urine
where inoculated on CLED (Cysteine Lactose
Electrolyte Deficient agar) and incubated at 37°C
for 24 hours. The suspected isolates were identified
based on Gram staining, cultural and biochemical
characters.
Determination of Antibiotic Susceptibility and Screening
for MRSA: The Kirby-Bauer’s disk diffusion method was
used to check the antibiotics susceptibility pattern of
isolated MRSA. Sterile swabs were used to pick the
inoculums and streaking was done over the entire
sterile surface of Mueller Hinton agar plate. The streaking
was repeated 2-3 times by rotating the plate each
time to ensure the uniform distribution of inoculums.
The antimicrobial disks of specific concentration were
dispensed onto the medium surface gently using sterile
forceps. Each disk was pressed down enough to come in
contact with agar surface and was incubated for 24 hours
at 37°C. Zones around the antibiotics disk were measured.
A clear zone indicates that bacteria are sensitive to that
antibiotic while absence of clear zone indicates resistance
of bacteria against particular antibiotic and results were
reported as per CLSI guidelines. Antibiotic such as
vancomycin, teicoplanin, linezolid, clindamycin, cefoxitin,
ceftazidine, ceftriaxone, vovobiocin, chloramphenicol
and amikacin.
Statistical Analysis: The analysis was done by using the
statistics software for windows.
RESULTS
In the present study among total isolates
(450), 320 (71.11%) were confirmed as MRSA.
When sample-wise distribution was checked it was
found that MRSA was more prevalent in pus 165
(51.56%), in blood 70 (28.17)%, in urine 59 (18.43%) and
MRSA was less prevalent in sputum sample 26 (8.12%)
(Table 1).
Antibiotic Susceptibility Profile of MRSA: All MRSA
isolates were showed complete resistance to amikacin and
penicillin while complete sensitive to vancomycin,
linezolid and teicoplanin. These MRSA strains were also
showed various degree of resistance to other
antimicrobials such as Clindamycin (46.87%), cefoxitin
(47.54%), ceftazidine (53.12%), ceftriaxone (59.06%) and
chloramphenical (55.55%) (Table 2).
 World J. Zool., 10 (4): 318-322, 2015

Table 1: Sample-wise distribution of MRSA which is due to the infection control program and strict
S.NO Samples Frequency Of MRSA, N (%)
ascetic techniques [21].
1 Pus 165 (51.56)
2 Blood 70 (28.17)
In our study maximum numbers of MRSA (51.56%)
3 Urine 59 (18.43) were isolated from pus. A similar study was carried out in
4 Sputum 26(8.12) India that reported the same prevalence rate of MRSA in
Total 320 pus samples [22]. The main reasons may be the increased
number of pus specimen compared to other samples
Table 2: Antibiotic susceptibility pattern against MRSA
received in our bacteriology section. Residence in a care
Intermediae Resistivity
S # Antibiotics Sensitivity n% sensitivity n% n% facility for a long time, catheters, dialysis and other
1 Vancomycin 320(100%) 0 (00) 0 (00) medical devices also contribute as a risk factor of MRSA.
2 Teicoplanin 320(100%) 0 (00) 0 (00) Furthermore, busy surgeons and paramedical staff also
3 Linezolid 320(100%) 0 (00) 0 (00) contribute to this scenario. However, some cases of
4 Clindamycin 170(53.12%) 0 (00) 150(46.87%)
5 Cefoxitin 153(47.81%) 4.68(15.66%) 152(47.54%)
MRSA infection were also reported from healthy
6 Ceftazidine 124(38.75%) 26(8.12%) 170(53.12%) communities without having any risk factor for MRSA
7 Ceftriaxone 119(45.46%) 12(3.75%) 189(59.06%) [23]. MRSA is also resistant to all other group members of
8 Novobiocin 80.31% 20.66% 0 (00) beta-lactam antibiotics including penicillins,
9 Chloramphenical 145(45%) 0 (00) 275(55.55%)
10 Ciprofloxacin 50% 25% 25%
cephalosporins and cephamycins. Along with that MRSA
11 Penicillin 0 (00) 0 (00) 320(100%) is often resistant to other classes of antimicrobials
12 Amikacin 0 (00) 0 (00) 320(100%) agents, like aminoglycosides, quinolones and macrolides.
This feature makes MRSA as multidrug-resistant bacteria
DISCUSSION [24]. The progressive spread of MRSA poses a huge
threat to the patients as well as to the community in term
MRSA is recognized as a major threat to the patients of diseases and high financial losses. The high variation
globally and their associated infection is a challenge for in number of MRSA among various hospital setting
clinicians due to its rapid spread and limited therapeutic limited the therapeutic option [24]. Regarding other
options available [13] As soon as MRSA introduced, therapeutic options of MRSA all isolates were found
steady rise in the number of S .aureus isolates was found uniformly sensitive to Vancomycin and Linezolid, thus
in various clinical settings. Several studies have been making treatment options possible. Similar findings was
carried out to find MRSA prevalence in different observed by Ahmad et al., in Saudi Arabia while
infections. Study that reported by National Nosocomial performing a prevalence study to find out nosocomial
Infections surveillance was showed an increase of MRSA infections of MRSA in worker of a hospital [25]. In this
range from 2.5% to 29% in 1975 to 1991 [14]. Ashiq and study resistance to pencillin, amikacin, ceftriaxone
Tareen in a prospective study reported 5% prevalence clindamycin, chloramphinicol, ceftazidine cefoxitin and
of MRSA in Karachi [15]. Bukhari et al. [16] in 2004 ciprofloxacin suggest that the use of these antibiotics
found out that 38.5% of bacterial isolates were MRSA, should be carefully prescribed to treat MRSA infections.
in an antibiotic susceptibility based study conducted
at King Edward Medical College, Lahore, Pakistan. CONCLUSION
Similarly, Khatoon et al. [17] concluded 38.5%
prevalence of MRSA in a laboratory based antibiotic MRSA is prevalent in every nook and corner of our
susceptibility study, carried out from June 2000 to hospital. It is the most important nosocomial pathogen
December 2000. and an alarming superbug, posing a serious threat to
Similar studies were also carried out in different parts infection control program. Strict surveillance, timely
of the world like Germany, France, Spain, Italy and the diagnosis and effective control measures are urgently
United Kingdom which revealed about 25% MRSA needed to check its rapid spread. Our hospital direly
prevalence while Austria, Poland, Slovakia, Czech needs effective and prompt control measures to reduce
Republic had reported MRSA rates of 7% to 14%. [18] morbidity, mortality and economy loss due to MRSA.
Karakatsanis et al. [19] reported 40% MRSA prevalence
in Greece. While George reported a high prevalence (73%) REFERENCES
of MRSA based on bacteriological, epidemiological and
clinical observation, in a Greek hospital [20]. But studies 1. Taubes, G., 2008. The bacteria fight back. Science
from the Westerns countries show a decline of MRSA 321: 356-361.

320
 World J. Zool., 10 (4): 318-322, 2015
2. Chongtrakool, P., T. Ito, X. Ma, Y. Kondo and
S. Trakulsomboon, 2006. Staphyloccoal cassette
chromosome mec (SCCmec) typing of Methicillin
resistance Staphyloccus aureus strains isolated in 11
Asian countries: A proposal for a new nomenclature
for SCCmec elements. Antimicrobial Agents and
Chemotherapy, 50: 1001-1012.
3. Archana, P. Iyer, Ibtisam, Baghallab, A. Mai and
K. Taha, 2014. Nosocomial Infections in Saudi Arabia
Caused by Methicillin Resistance Staphylococcus
aureus (MRSA), Clin Microbial, 3: 3.
4. Al Tawfiq., 2006. Incidence and Epidemiology of
Methicillin-Resistant Staphylococcus aureus
Infection in a Saudi Arabian Hospital. Infection
Control and Hospital Epidemiology, 27: 1137-1139.
5. Fact sheet on: Methicillin Resistant Staphylococcus
aureus, Last update Jan. 2011, The center for Food
Security & Public Health, institute for international
cooperation in animal biologics, Iowa State
University, College of Veterinary Medicine.
6. Pantosti, A., A. Sanchini and M. Monaco, 2007.
Mechanisms of antibiotic resistance in
Staphylococcus aureus. Future Microbiol.,
2: 323-334.
7. Al, Tawfiq, 2006. Incidence and Epidemiology of
Methicillin-Resistant Staphylococcus aureus
Infection in a Saudi Arabian Hospital. Infection
Controland Hospital Epidemiology, 27: 1137-1139.
8. Fuda, C., M. Suvorov, S.B. Vakulenko and
S. Mobashery, 2004. The basis for resistance to beta-
lactam antibiotics by penicillin binding protein 2a of
methicillin-resistant Staphylococcus aureus. J. Biol.
Chem., 279: 40802e6.
9. Kollef, M.H., S. Ward, G. Sherman, D. Prentice and
R. Schaiff, 2000. In adequate treatment of nosocomial
infections is associated with certain empiric antibiotic
choices. Crit Care. Med., 28: 3456-3464.
10. Cross, J.T. and G.D. Campbell, 1999. Drug-resistant
pathogens in community- and hospital-acquired
pneumonia. Clin Chest Med., 20: 502.
11. Stryjewski, M.E. and G.R. Corey, 2014. Methicillin-
resistant Staphylococcus aureus: an evolving
pathogen. Clin Infect Dis., 58: 10-9.
12. De, L.C.R., B. Johnson and D. Christensen, 2004.
Emergence of S. aureus as etiology of bacteremic
community-acquired pneumonia. Results from the
Community Acquired Pneumonia Organization
(CAPO) International Study. Poster Abstracts.
ICCAC Scientific Meeting 2004.
321
13. Ashiq, B. and A.K. Tareen, 1989. Methicillin
Resistant S. aureus in a teaching hospital of Karachi-
a Laboratory study. J Pak Med Assoc., 39: 6-9.
14. Bukhari, A. Iqbal, N. Khatoon, N. Iqbal, S. Naeem and
G.R. Qureshi, 2004. A laboratory study of
susceptibility of Methicillin resistant S. aureus
(MRSA). Pak J. Med. Sci., 20: 229-33.
15. Khatoon, N., M.H. Bukhari, J.R. Riaz, A.S. Sheikh,
A. Iqbal and S. Naeem, 2002. Prevalence of
Methicillin Resistant S. aureus (MRSA) infection
laboratory study at Mayo Hospital Lahore.
Biomedica., 18: 49-52.
16. Eckmann, C. and M. Dryden, 2010. Treatment of
complicated skin and soft-tissue infections
caused by resistant bacteria: value of linezolid,
tigecycline, daptomycin and vancomycin. Eur J. Med.
Res., 15: 554-63.
17. Polychonopoulou, K., C. Arakatsanis, D. Petropolou
and P. Gargalicianos, 1987. Bacterial resistance of
Staphylococci in a Greek Hospital. Bacteriological,
clinical and epidemiological observations In: Ist
International Conference of Hospital Infection
Society, London (Abstract), pp: 1-3.
18. George, R.C., B.R. Marpoles and I.C. Bakk, 1987.
Susceptibility of recent hospital isolates of S. aureus
to 20 antimicrobial agents. In: Ist International
Conference of Hospital Infection Society, London
(Abstract), pp: 11-4.
19. Mulligan, M.E., Murray-LKA. and B.S. Ribner, 1993.
Methicillin resistant Staphylococcus aureus: a
consensus review of the microbiology, pathogenesis
and epidemiology with implications for prevention
and management. Am. J. Med., 94: 313-28.
20. Mehta, A.P., C. Rodrigues, K. Sheth, S. Jani,
A. Hakimiyan and N. Fazalbhoy, 1998. Control of
methicillin resistant Staphylococcus aureus in a
tertiary care Centre A five year study. Ind J. Med.
Microbiol., 16: 31-4.
21. Naimi, T.S., K.H. LeDell, D.J. Boxrud, A.V. Groom,
C.D. Steward, S.K. Johnson, et al., 2000.
Epidemiology and clonality of community-acquired
methicillin resistant Staphylococcus aureus in
Minnesota, 1996-1998. Clin Infect Dis., 33: 990-6.
22. Van, H.S.J., D. Stark, B. Lockwood, D. Marriott and
J. Harkness, 2007. Methicillin-resistant
Staphylococcus aureus (MRSA) detection:
comparison of two molecular methods (IDI-MRSA
PCR assay and GenoType MRSA Direct PCR assay)
with three selective MRSA agars (MRSA ID,
MRSASelect and CHRO Magar MRSA) for use
with infection-control swabs. J. Clin Microbiol.,
45: 2486-90.
 World J. Zool., 10 (4): 318-322, 2015

23. Kim, T., P.I. Oh and A.E. Simor, 2001. The economic 24. Ahmad, S., 2007. The prevalence of Staphylococcus
impact of methicillin resistant Staphylococcus aureus colonization among Healthcare Workers of a
aureus in Candian hospitals. Infect Control Hosp Specialist Hospital in Saudi Arabia. J. Clin Diag Res.,
Epidimiol., 22: 99-104. 4: 2438-41.

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