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Phage typing of methicillin-resistant Staphylococcus aureus

Abu Hanifah Y

Malays J Pathol, 1990 Dec;12(2):107-9.
PMID: 2102965

448 isolates of methicillin-resistant Staphylococcus aureus (MRSA) from clinical specimens of patients from the University Hospital, Kuala Lumpur, were phage-typed. These included 35 strains causing two separate outbreaks of infection, one in surgical Ward 6B and another in the Special Care Nursery (SCN). Antibiograms of these outbreak strains in Ward 6B and SCN were entirely different. Phage-typing revealed that 72% of the MRSA isolates were typable. They were typed entirely by Group III phages, the majority (76%) of which were phage type 85. There was only one isolate in SCN which was typed by Group I (phage 80) and Group III phages. None were typed by phages 94, 95, 96 and Group II phages. 14.6% of the typable isolates gave the long pattern reaction of the phage 6/47/54/75/77/83A/84/85 complex. The majority of the outbreak strains in Ward 6B were of phage type 85, whereas those in the SCN were all of the 6/47/54/75/77/83A/84 phage pattern with the exception of one isolate which was also typed by phage 80, a Group I phage.

Matched MeSH terms: Methicillin Resistance*
Fulltext DNA fingerprinting of methicillin-resistant Staphylococcus aureus (MRSA) by pulsed-field gel electrophoresis (PFGE) in a teaching hospital in Malaysia

Alfizah H, Norazah A, Nordiah AJ, Lim VKE

Med J Malaysia, 2002 Sep;57(3):319-28.
PMID: 12440272 MyJurnal

Methicillin-resistant Staphylococcus aureus (MRSA) has been prevalent in our hospital over the last three years. Differentiation among MRSA strains by DNA typing in addition to antibiotic resistance pattern surveillance is crucial in order to implement infection control measures. The aim of this study was to characterize MRSA isolates from patients admitted to Hospital Universiti Kebangsaan Malaysia (HUKM) by phenotypic (analyses of antibiotic susceptibility pattern) and genotypic (PFGE) techniques to determine the genetic relatedness of the MRSA involved and to identify endemic clonal profiles of MRSA circulating in HUKM. Seventy one MRSA strains collected between January to March 2000 from patients from various wards in HUKM were tested for antimicrobial resistance and typed by pulsed-field gel electrophoresis (PFGE). Four major types of PFGE patterns were identified (A, B, C and D) among MRSA strains. Two predominant PFGE types were recognised, Type A (59.2%) and Type B (33.8%). Most of these strains were isolated from ICU, Surgical wards and Medical wards. MRSA strains with different PFGE patterns appeared to be widespread among wards. Strains with the same antibiotype could be of different PFGE types. Most of isolates were resistant to ciprofloxacin, erythromycin, gentamicin and penicillin. One isolate with a unique PFGE pattern Type D and susceptible to gentamicin was identified as a different clone. Some isolates obtained from the same patient showed different PFGE subtypes suggesting that these patients were infected/colonized with multiple MRSA strains. PFGE analysis suggests that MRSA strains with different PFGE types was propagated within our hospital. The relationship between antibiotic susceptibility and PFGE patterns was independent. The ability of PFGE technique in differentiating our MRSA strains make it a method of choice for investigating the source, transmission and spread of nosocomial MRSA infection, and thus an appropriate control programme can be implemented to prevent the spread of MRSA infection.

Matched MeSH terms: Methicillin Resistance/genetics*
Lactoferrin modulates the biofilm formation and bap gene expression of methicillin-resistant Staphylococcus epidermidis

Khanum R, Chung PY, Clarke SC, Chin BY

Can J Microbiol, 2023 Feb 01;69(2):117-122.
PMID: 36265186 DOI: 10.1139/cjm-2022-0135

Lactoferrin is an innate glycoprotein with broad antibacterial and antibiofilm properties. The autonomous antibiofilm activity of lactoferrin against Gram-positive bacteria is postulated to involve the cell wall and biofilm components. Thus, the prevention of biomass formation and eradication of preformed biofilms by lactoferrin was investigated using a methicillin-resistant Staphylococcus epidermidis (MRSE) strain. Additionally, the ability of lactoferrin to modulate the expression of the biofilm-associated protein gene (bap) was studied. The bap gene regulates the production of biofilm-associated proteins responsible for bacterial adhesion and aggregation. In the in vitro biofilm assays, lactoferrin prevented biofilm formation and eradicated established biofilms for up to 24 and 72 h, respectively. Extensive eradication of MRSE biofilm biomass was accompanied by the significant upregulation of bap gene expression. These data suggest the interaction of lactoferrin with the biofilm components and cell wall of MRSE, including the biofilm-associated protein.

Matched MeSH terms: Methicillin Resistance/genetics
Fulltext Methicillin-resistant Staphylococcus aureus (MRSA)--phage-typing of Malaysian and international isolates

Hanifah YA

Singapore Med J, 1991 Dec;32(6):438-40.
PMID: 1788605

Twenty-one isolates of methicillin-resistant Staphylococcus aureus (MRSA) from Malaysia (M-MRSA) derived from various sources associated with nosocomial infections were phage-typed and compared with 54 international isolates associated with epidemic and sporadic episodes of infections. It appeared that the majority of M-MRSA were non-typable by the international basic set of phages. Two (9.5%) were typed by phage 85. Phage-typing of MRSA revealed that the strains were almost completely restricted to phage groups III and a lesser portion to phage groups I and III.

Matched MeSH terms: Methicillin Resistance*
Fulltext Toxin gene profiling of methicillin-susceptible Staphylococcus au

Hui-min, Neoh, Saberi Saimun, Hassriana Fazilla Sapr, Salasawati Hussin, Rahman Jamal

Asia-Pacific Journal of Molecular Medicine, 2017;7(2):1-3.
MyJurnal

Entero- and exfoliative toxin gene profiling of 237 methicillin-susceptible Staphylococcus aureus (MSSA) isolated
from Universiti Kebangsaan Malaysia Medical Centre (UKMMC) were carried out via PCR amplification. Among
the tested toxin genes, sei was found to be the most prevalent (54.9%).

Matched MeSH terms: Methicillin Resistance
Fulltext Isolation and Characterization of Antibacterial Compounds from Ganoderma boninense Against Methicillin-Resistant Staphylococcus aureus (MRSA)

Syahriel Abdullah, Ling, Yee Soon, Daim, Sylvia Jerome, Chong, Khim Phin

Borneo Journal of Medical Sciences, 2018;12(101):11-12.
MyJurnal

Community and nosocomial-associated methicillin-resistant Staphylococcus aureus (MRSA) infections in Malaysian healthcare setting are terrifically increasing in recent years. There is an urgent need for an effective antibacterial agent to cope with this important issue. Due to the development of new antibiotics is not parallel with the increase of cases of infections, researchers have initiated isolation of compounds from natural resources to develop new potent therapeutic agent. Meanwhile, Ganoderma boninense is an oil palm devastating pathogen, which has been known to contain many bioactive compounds that might be potential to be developed as a new source of therapeutic agent.

Matched MeSH terms: Methicillin Resistance
Fulltext Antibacterial activity of leaf extracts of Baeckea frutescens against methicillin-resistant Staphylococcus aureus

Razmavar S, Abdulla MA, Ismail SB, Hassandarvish P

Biomed Res Int, 2014;2014:521287.
PMID: 25028658 DOI: 10.1155/2014/521287

This study was based on screening antibacterial activity of the ethanol extract of Baeckea frutescens L. against MRSA clinical isolates, analyzes the potential antibacterial compound, and assesses the cytotoxicity effect of the extract in tissue culture. Leaves of Baeckea frutescens L. were shade dried, powdered, and extracted using solvent ethanol. Preliminary phytochemical screening of the crude extracts revealed the presence of alkaloids, flavonoids, steroids, terpenoids, phenols, and carbohydrates. The presence of these bioactive constituents is related to the antibacterial activity of the plant. Disc diffusion method revealed a high degree of activity against microorganisms. The results confirm that Baeckea frutescens L. can be used as a source of drugs to fight infections caused by susceptible bacteria.

Matched MeSH terms: Methicillin Resistance/drug effects*
Simultaneous species identification and detection of methicillin resistance in staphylococci using triplex real-time PCR assay

Sabet NS, Subramaniam G, Navaratnam P, Sekaran SD

Diagn Microbiol Infect Dis, 2006 Sep;56(1):13-8.
PMID: 16650954

For rapid identification of methicillin-resistant Staphylococcus aureus, molecular methods are generally targeting mecA and species-specific genes. Sa442 DNA fragment is a popular species-specific target. However, recently, there have been few reports on S. aureus isolates that are negative for Sa442 fragment; therefore, use of single gene or DNA-fragment-specific polymerase chain reaction (PCR) for identification of microbial isolate may result in misidentification. This study includes CoA gene in parallel with Sa442 marker for identification of S. aureus. This further improves the specificity of the assay by checking for 2 determinants simultaneously for the identification of S. aureus and can prevent misidentification of S. aureus isolates lacking Sa442 DNA fragment. In this study, the newly developed triplex real-time PCR assay was compared with a quadruplex conventional gel-based PCR assay using the same primer sets in both assays. The dual-labeled TaqMan probes (ProOligo, France) for these primers were specifically designed and used in a real-time PCR assay. The clinical isolates (n = 152) were subjected to both PCR assays. The results obtained from both assays proved that the primer and probe sets were 100% sensitive and 100% specific for identification of S. aureus and detection of methicillin resistance. This triplex real-time PCR assay represents a rapid and powerful method for S. aureus identification and detection of methicillin resistance.

Matched MeSH terms: Methicillin Resistance/genetics*
Methicillin-resistant Staphylococcus aureus bacteraemia at a tertiary teaching hospital

Cheong I, Samsudin LM, Law GH

Br J Clin Pract, 1996 Jul-Aug;50(5):237-9.
PMID: 8794598

Between July and December 1994, 25 patients with MRSA bacteraemia were treated at the Hospital Kuala Lumpur, a tertiary hospital in Malaysia with 3000 beds. The patients included 15 males and 10 females whose mean age was 46.7 years (range 13-75). The sources of their MRSA were: Urology/Nephrology, 11; General ICU, six; Orthopaedic, four; Medicine, three; Surgery, one. Their underlying diseases were: end-stage and chronic renal failure, 11; burns, three; acute necrotising pancreatitis, two; haematological malignancies, two; and one each of fracture of the neck of the femur, pustular psoriasis, alcoholic cirrhosis, liver abscess, peptic ulcer (antrectomy), choledochol cyst, and abdominal aneurysm with gangrene of the legs. Six patients were also diabetic. A total of 19 infections were considered nosocomial. The duration of hospital stay ranged from one to 60 days, mean 16 days. On the day of blood culture, 20 patients (80%) were febrile and 15(60%) had leucocytosis. A total of 14 patients were considered to have received prolonged broad-spectrum antibiotics before the bacteraemia; of these, 11 had had either a third-generation cephalosporin and/or a quinolone. The primary foci of infection were: vascular access dialysis catheters, six; infected AV fistulae, three; non-surgical wounds, five; orthopaedic pin, one; multiple venous lines and catheters, nine; unknown, one. The sensitivities to anti-MRSA antibiotics were: vancomycin, 100%; fusidic acid, 96%; rifampicin, 96%; ciprofloxacin and perfloxacin 28% each. In all, 13 patients (52%) eventually died; nine of these deaths were directly attributed to MRSA bacteraemia. The microbiological eradication rate was 88%. Mortality was significantly associated with duration of hospital stay and failure to remove the infected catheters/peripheral lines after the development of MRSA bacteraemia.

Matched MeSH terms: Methicillin Resistance*
Pulsed-field gel electrophoresis of chromosomal DNA of methicillin-resistant Staphylococcus aureus associated with nosocomial infections

Hanifah YA, Hiramatsu K

Malays J Pathol, 1994 Dec;16(2):151-6.
PMID: 9053564

Methicillin-resistant Staphylococcus aureus (MRSA) infection has been endemic in the University Hospital, Kuala Lumpur since the late 1970s. Fifty isolates of MRSA obtained from clinical specimens of patients with nosocomial infections associated with this organism have been studied by pulsed-field gel electrophoresis (PFGE) of its chromosomal DNA fragments to discrimate between strains and to identify the predominant strain. Twenty-one chromosomal patterns were observed which could be further grouped into nine types. The predominant strain was Type 9-b (40% of isolates) found mainly in the Orthopaedic and Surgical Units. Outbreak strains found in the Special Care Nursery were of Type 1, entirely different from those of the surgical ward S2, which were of Type 9-b. Type 8 strains were found mainly at one end of the hospital building where the maternity, paediatric and orthopaedic units were situated. Genomic DNA fingerprinting by PFGE is recommended as a useful and effective tool for the purpose of epidemiological studies of MSRA infections, particularly for nosocomial infections.

Matched MeSH terms: Methicillin Resistance/genetics*
Pyomyositis revisited

Kong NC, Asmah J, Lim VK, Ong PH, Adam PA

Ann Acad Med Singap, 1996 Jul;25(4):609-11.
PMID: 8893941

Pyomyositis, purportedly a common tropical infection affecting mainly healthy adults and children, appears to be most uncommon in this region. We report a case of pyomyositis caused by a Methicillin-resistant Staphylococcus aureus (MRSA) in a previously healthy army officer. This case serves to illustrate the difficulty in recognising this disease entity, which is why many cases may have been missed. With the increasing incidence of MRSA nosocomial infections, the emergence of MRSA in a hitherto community-acquired infection poses a major concern especially since intravenous drug abuse and acquired immune deficiency syndrome (AIDS) are on the rise in our country. We hope to inculcate greater awareness of this infection.

Matched MeSH terms: Methicillin Resistance*
Detection of mecA and ermA genes and simultaneous identification of Staphylococcus aureus using triplex real-time PCR from Malaysian S. aureus strain collections

Sabet NS, Subramaniam G, Navaratnam P, Sekaran SD

Int J Antimicrob Agents, 2007 May;29(5):582-5.
PMID: 17314034

A triplex real-time polymerase chain reaction (PCR) assay was used for the simultaneous detection of mecA (methicillin resistance), ermA (erythromycin resistance) and femA (Staphylococcus aureus identification) genes in a single assay. Among 93 clinical S. aureus hospital isolates, there were 48 methicillin-resistant S. aureus (MRSA) and 45 methicillin-sensitive S. aureus (MSSA) isolates. Screening the isolates using the triplex real-time PCR assay, the mecA, ermA and femA genes were detected in all MRSA isolates. The triplex real-time PCR assay was completed within 3h and is a useful genotypic method for detecting the resistance determinants as well as for the identification of S. aureus isolates. These findings will assist the clinical laboratory in identifying these resistance genes and S. aureus rapidly, thus benefiting patient therapy. This study represents a valuable source of information for researchers to study the local antibiotic resistance pattern, which can increase our knowledge of the antibiotic resistance profile, using real-time PCR technology.

Matched MeSH terms: Methicillin Resistance/genetics
First community-acquired meticillin-resistant Staphylococcus aureus in Malaysia

Nor Shamsudin M, Sekawi Z, van Belkum A, Neela V

J Med Microbiol, 2008 Sep;57(Pt 9):1180-1181.
PMID: 18719195 DOI: 10.1099/jmm.0.47844-0
Matched MeSH terms: Methicillin Resistance*
Fulltext DNA fingerprinting of methicillin-resistant Staphylococcus aureus by pulsed-field gel electrophoresis (PFGE): comparison of strains from 2 Malaysian hospitals

Norazah A, Liew SM, Kamel AG, Koh YT, Lim VK

Singapore Med J, 2001 Jan;42(1):15-9.
PMID: 11361232

To determine and compare the pulsed-field gel electrophoresis (PFGE) patterns of endemic MRSA strains in 2 major Malaysian hospitals and to compare the PFGE patterns with antibiotypes of the strains studied.

Matched MeSH terms: Methicillin Resistance/genetics*
Fulltext An outbreak of methicillin-resistant Staphylococcus aureus in a neonatal intensive care unit in Singapore: a 20-month study of clinical characteristics and control

Tan KW, Tay L, Lim SH

Singapore Med J, 1994 Jun;35(3):277-82.
PMID: 7997904

Methicillin resistant Staphylococcus aureus (MRSA) is a major infection control problem in many countries. There have been many reports of outbreaks in neonatal nurseries including, in our part of the world, Australia, Hong Kong and Malaysia. A recent outbreak of MRSA in the neonatal intensive care unit in the Kandang Kerbau Hospital, Singapore, presented us with the opportunity to study the clinical characteristics of the outbreak and the effects of infection control measures. Neonates admitted to the neonatal intensive care unit were studied over a 20-month period. They were all screened for nasal colonisation on admission and weekly thereafter. Infections were documented. Over this period there were altogether 2,576 admissions of which 85 infants had nasal colonisation with MRSA (3.3%) and 28 developed infections (1%). Although the majority of infants colonised by MRSA suffered no ill effects, 3 had septicaemia and 2 had septicaemia with osteomyelitis. There were no deaths. Standard infection control measures with barrier nursing and the use of mupirocin nasal ointment were ineffective, and control was achieved only after strict cohorting together with the use of mupirocin was instituted. This was done without additional costs to the department and without additional nurses or doctors.

Matched MeSH terms: Methicillin Resistance*
Survey of Staphylococcus isolates among hospital personnel, environment and their antibiogram with special emphasis on methicillin resistance

Shobha KL, Rao PS, Thomas J

Indian J Med Microbiol, 2005 Jul;23(3):186-8.
PMID: 16100427

The objective of this study was to find the prevalence of Staphylococcus spp. carriage among hospital personnel and hospital environment and their antibiogram with special emphasis on methicillin resistance. A total of 205 samples from hospital personnel and environment were collected from casualty, oncology and multidisciplinary cardiac unit ward of Kasturba Medical College Hospital, Manipal. Samples were collected using sterile cotton wool swabs and inoculated into brain heart infusion broth. Subcultures were done onto blood agar and MacConkey's agar. Isolates were identified by standard methods up to species level. Antimicrobial susceptibility test was performed according to standardized disc diffusion Kirby-Bauer method. Each of the isolates was screened for methicillin resistance using oxacillin disc on Mueller Hinton agar plate followed by MIC for methicillin and cefoxitin susceptibility test by disc diffusion method. Sixty five out of 205 strains (31.7%) were Staphylococcus spp. and all of them were coagulase negative. Most of the strains belonged to S.epidermidis 49.23% (32/65) followed by S. saprophyticus 26.15% (17/65). Maximum isolates of S.epidermidis were from anterior nares 28.12% (9/32 strains of S.epidermidis). Highest number of methicillin resistant coagulase negative strains (3/9, 33.33%) were isolated from stethoscope of multidisciplinary cardiac unit ward followed by carriers in the anterior nares (2/9, 22.22%). Methicillin resistant coagulase negative staphylococci are prevalent in anterior nares of hospital personnel and in the hospital environment thereby providing a definite source for hospital acquired infection. All isolates were sensitive to vancomycin, ciprofloxacin and amikacin.

Matched MeSH terms: Methicillin Resistance*
Fulltext Massive Tibial Bone Regeneration with Autologous Peripheral Blood Stem Cells using Ilizarov Bone Transport: A Case Report

Saw KY, Gill R, Low TC

Malays Orthop J, 2020 Nov;14(3):166-169.
PMID: 33403079 DOI: 10.5704/MOJ.2011.026

This is a case report of a Gustilo-Anderson Type IIIB comminuted open right tibial fracture with massive bone loss, complicated by methicillin-resistant Staphylococus aureus (MRSA) infection. Non-viable and contaminated bony fragments were removed and infected bone resected. Soft tissue coverage and antibiotics were effective against the MRSA infection. A unifocal bone transport with the Ilizarov method regenerated 13cm of the missing tibia. Autologous peripheral blood stem cells (PBSC) injections into the osteogenesis site boosted bone regeneration and consolidation with a shortened Bone Healing index (BHI) of 23 days/cm.

Matched MeSH terms: Methicillin Resistance
Antibacterial Action of Curcumin against Staphylococcus aureus: A Brief Review

Teow SY, Liew K, Ali SA, Khoo AS, Peh SC

J Trop Med, 2016;2016:2853045.
PMID: 27956904

Curcumin, the major constituent of Curcuma longa L. (Zingiberaceae family) or turmeric, commonly used for cooking in Asian cuisine, is known to possess a broad range of pharmacological properties at relatively nontoxic doses. Curcumin is found to be effective against Staphylococcus aureus (S. aureus). As demonstrated by in vitro experiment, curcumin exerts even more potent effects when used in combination with various other antibacterial agents. Hence, curcumin which is a natural product derived from plant is believed to have profound medicinal benefits and could be potentially developed into a naturally derived antibiotic in the future. However, there are several noteworthy challenges in the development of curcumin as a medicine. S. aureus infections, particularly those caused by the multidrug-resistant strains, have emerged as a global health issue and urgent action is needed. This review focuses on the antibacterial activities of curcumin against both methicillin-sensitive S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA). We also attempt to highlight the potential challenges in the effort of developing curcumin into a therapeutic antibacterial agent.

Matched MeSH terms: Methicillin Resistance
spa diversity of methicillin-resistant and -susceptible Staphylococcus aureus in clinical strains from Malaysia: a high prevalence of invasive European spa-type t032

Jones SU, Chua KH, Chew CH, Yeo CC, Abdullah FH, Othman N, et al.

PeerJ, 2021;9:e11195.
PMID: 33889447 DOI: 10.7717/peerj.11195

Background: Staphylococcus aureus is one of the important pathogens causing nosocomial infection. spa typing allows identification of S. aureus clones in hospital isolates and is useful for epidemiological studies and nosocomial infection control. This study aims to investigate the spa types in Malaysian S. aureus isolates obtained from various clinical specimens.
Method: A total of 89 methicillin-resistant S. aureus (MRSA) [pus (n = 55), blood (n = 27), respiratory (n = 5), eye (n = 2)] isolates and 109 methicillin-susceptible S. aureus (MSSA) [pus (n = 79), blood (n = 24), respiratory (n = 3), eye (n = 2) and urine (n = 1)] isolates were subjected to spa typing with sequences analysed using BioNumerics version 7.
Results: The spa sequence was successfully amplified from 77.8% of the strains (154/198) and 47 known spa types were detected. The distribution of known spa types in MRSA (36.2%, 17/47) was less diverse than in MSSA (70.2%, 33/47). The most predominant spa types were t032 (50%) in MRSA, and t127 (19%) and t091 (16.7%) in MSSA, respectively. spa type t091 in MSSA was significantly associated with skin and soft tissue infections (p = 0.0199).
Conclusion: The previously uncommon spa type t032 was detected in the Malaysian MRSA strains, which also corresponded to the most common spa type in Europe and Australia, and has replaced the dominant spa type t037 which was reported in Malaysia in 2010.

Matched MeSH terms: Methicillin Resistance
Characterization of resistance to selected antibiotics and Panton-Valentine leukocidin-positiveStaphylococcus aureusin a healthy student population at a Malaysian University

Suhaili Z, Rafee P', Mat Azis N, Yeo CC, Nordin SA, Abdul Rahim AR, et al.

Germs, 2018 Mar;8(1):21-30.
PMID: 29564245 DOI: 10.18683/germs.2018.1129

Introduction: This study aims to assess the antimicrobial susceptibility profiles ofStaphylococcus aureusstrains isolated from university students and to determine the prevalence of constitutive and inducible clindamycin resistance, the latter being able to cause therapeutic failure due to false in vitro clindamycin susceptibility.
Methods: S. aureus
strains were isolated from the nasal swabs of 200 health sciences students of a Malaysian university. Twelve classes of antibiotics were used to evaluate the antimicrobial susceptibility profiles with the macrolide-lincosamide-streptogramin B (MLSB) phenotype for inducible clindamycin resistance determined by the double-diffusion test (D-test). Carriage of resistance and virulence genes was performed by PCR onS. aureusisolates that were methicillin resistant, erythromycin resistant and/or positive for the leukocidin gene,pvl(n=15).
Results: Forty-nine isolates were viable and identified asS. aureuswith four of the isolates characterized as methicillin-resistantS. aureus(MRSA; 2.0%). All isolates were susceptible to the antibiotics tested except for penicillin (resistance rate of 49%), erythromycin (16%), oxacillin (8%), cefoxitin (8%) and clindamycin (4%). Of the eight erythromycin-resistant isolates, iMLSBwas identified in five isolates (three of which were also MRSA). The majority of the erythromycin-resistant isolates harbored themsrAgene (four iMLSB) with the remaining iMLSBisolate harboring theermCgene.
Conclusion: The presence of MRSA isolates which are also iMLSBin healthy individuals suggests that nasal carriage may play a role as a potential reservoir for the transmission of these pathogens.

Matched MeSH terms: Methicillin Resistance

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