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Articles by E. Amghalia
Total Records ( 3 ) for E. Amghalia
  E. Amghalia , Nagi AL-Haj , Mariana N. Shamsudin , V. Neela and W.M.Z. Somarny
  Staphylococcus aureus is among the most prominent pathogen in both community acquired and nosocomial infection. The epidemiology analysis of Staphylococcus aureus isolates will be required to ascertain the incidence, prevalence and diversity of strains. To investigate the epidemiological of S. aureus in Malaysia, a highly reliable typing method-Randomly amplified polymorphic DNA was applied to 50 S. aureus isolates obtained from 3 different hospitals in Malaysia namely Hospital Tunku Ampuan Afzan Kuantan, Hospital Besar Seremban and Hospital Miri Malaysia. The results obtained from this study showed that the isolates can be clustered into 8 different clones. All members of the respective clones are of the same origin. In addition, there were 2 clonal grouping of isolates for each hospital. However, the clonal groupings are not in accordance to the geographical distribution. To understand the epidemiology of these isolates in depth it is very important to have information about the patient’s history. The Nei and Li’s genetic distances obtained from this study ranged from 0.0803922-0.11111. Two genetic markers a band of size 500 bp when amplified with primer OPAE-14 and another marker band of size 750 bp amplified with primer OPAE-15 was identified and this band can be used as diagnostic marker for the rapid identification of S. aureus. Apart from the genetic markers, an epidemiological marker of 1200 bp was also identified for the Miri isolates. This marker can be used as the epidemiological marker for the identification of the isolates from Miri in the future outbreaks. From this study, RAPD has proved to be an useful aid to epidemiological investigations of S. aureus.
  E. Amghalia , Nagi A. AL-Haj , Mariana N. Shamsudin , Son Radu , Rozita Rosli , V. Neela and Raha A. Rahim
  Multiple drug resistant Staphylococcus aureus is one the most common nosocomial pathogen worldwide. The timely identification of this hospital acquired pathogen and detection of the various antibiotic resistant genes harbored is one of the most important function of the microbiology laboratory. In this study, we report the development of a multiplex PCR system for the diagnosis of S. aureus and the detection of clinically relevant antibiotic resistance genes harbored by some isolates. This system was designed to identify S. aureus at species level and to detect methicillin, gentamycin, erythromycin, vancomycin and mupirocin resistant genes, respectively from a single colony in a single tube reaction. All isolates amplified a 108 bp fragment (conserved in S. aureus) confirming the identity of S. aureus, 23 isolates produced a band at the position of 533 bp, 28 isolates at 139 bp and 30 isolates at 174 bp evidencing the presence of mecA (methicillin or oxacillin resistance), ermA (erythromycin resistance), aac (6`)-aph (2``) (gentamycin resistance) genes. None of the isolates amplified van A (vancomycin resistance) and ileS-2 (mupirocin resistance) genes showing the absence of their resistance in the isolates studied. These genotypic results when compared with classical antibiotic susceptibility tests showed less correlation. Overall, we found a correlation between phenotypic and genotypic methods of 60% for methicillin, 36.7% for gentamycin, 43.3% for erythromycin, 100% for vancomycin and mupirocin. This suggests that classical antibiotic sensitivity test is not accurate, but need to be supplemented with other methods to be applied in a clinical laboratory. The system developed in this study offers a rapid, simple specific and accurate detection of multiple antibiotic resistant genes in clinical S. aureus isolates and thus could be systematically applied as a diagnostic test in clinical microbiology laboratories, facilitating the design and use of antibiotic therapy.
  E. Amghalia , Nagi A. AL-Haj , Mariana N. Shamsudin , Nurmas I. Mashan , V. Neela and Zamberi Sekawi
  Methicilin Resistant Staphylococcus aureus (MRSA) implicated in many post-surgical and cancer treatment as well hospital and community fatalities need to be treated with an effective alternative antimicrobial agent. In the search for anti-MRSA agent, 2 types of natural products were investigated for inhibitory activity against MRSA. In addition to the bioassay, the activity of the anti-MRSA agent was elucidated based on the effect of both natural products on nucleotide changes of chromosome-encoded genes. In this study, the methanol extract of the red marine algae and the natural pure honey were studied for its antibacterial property based on disc diffusion test and Minimum Inhibitory Concentration (MIC). The effects of both natural products on selected gene sequences of S. aureus’s were determined by RT-PCR analysis. The genes of interest, which have been chosen in this study, are genes that are involved in the antibacterial mechanism including inhibition of cell wall synthesis, protein synthesis and nucleic acid synthesis. Five genes of interest chosen in this study include mecA gene, mecR1 gene, mecI gene, adaB gene and sav1017 gene. The results for antibacterial property showed the methanol extract of a red seaweed and the pure honey, inhibited growth of S. aureus strain according to the inhibition zones around discs saturated with the seaweed extract and pure honey, respectively. The MIC test showed decrease in growth of MRSA isolates after growing in broth incorporated with the extract and honey, respectively. The effect of the inhibitory activity of the natural products on selected gene sequences showed that several nucleotide changes occurred in the sequences of certain genes of interest based on the gene sequences of the cDNA after RT-PCR was carried out on the mRNA of S. aureus treated with the natural products. This research underlined that the inhibition effect of the natural products may be chromosome mediated evidenced by the changes of chromosome-encoded genes. The significant findings on activities of the seaweed extract and pure honey may become very useful in the process to find a better treatment for S. aureus infection especially, for the multiple drug resistant isolates. In addition, it is also, a new finding for natural product discovery through gene-expression analysis.
 
 
 
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