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Articles by Raha A. Rahim
Total Records ( 5 ) for Raha A. Rahim
  Nagi A. AL-Haj , Mariana N. Shamsudin , Raha A. Rahim and Zamri Ishak
  Although, PCR methods aimed on the detection of genes associated with the pathogenicity of Escherichia coli have been reported, tests allowing the direct identification of this serotype are rare. In this study the Random Amplified Polymorphic DNA (RAPD) fingerprinting technique allowed genetic diversity assessment of 25 E. coli isolates of various sources. A highly significant finding from the DNA fingerprinting is the display of a predominant band at a size of 308 bp when arbitrary OPAE-10 primer was used. After sequencing this fragment primer called secD was designed to be used as PCR primer. secD primer pairs was highly specific to detect all isolates including E. coli O157: H7.
  Nagi A. Al-Haj , Mariana N. Shamsudin , Raha A. Rahim and Zamri Ishak
  Quantitation assay of Escherichia coli, Samonell sp. and Vibrio cholerae cells investigated by exploiting the component consistently present on the outer surface of Gram-negative bacteria. In this study, a simple marine biolysate-based method for simultaneous detection of the gram negative pathogenic bacteria based on lipopolysaccharide component was optimized. The detection technique focused on the surface of these bacterial species, which is covered by polysaccharides and has high affinity to marine biolysate. E. coli, Samonell sp. and V. cholerae with similar initial cell count per mL have different but consistent absorbance readings by using the spectrophotometer and turbidity meter. This revealed that different genera of Gram Negative bacteria can be directly differentiated through standard curve that is plotted from the carbohydrate and marine biolysate assays absorbance readings. Both the assays elucidated a qualitative and quantitative detection of the pure culture pathogens.
  Nagi A. Al-Haj , Mariana N. Shamsudin , Raha A. Rahim and Zamri Ishak
  Recent advances in molecular techniques have revolutionized the detection of microorganism. The development of a molecular-based technique for detection of the three different targets of Enterbacteriacae was undertaken. Primer and probe were designed based on specific pepted of novel hemolymph protein of horseshoe crabs (Factor C anti-LPS) Tachypleus tridentatus that is believed to be involved in the binds to the lipopolysaccharide of Escherichia coli, Salmonella and Vibrio cholerae. The aim of our study the exploit part of cell wall polysaccharide in the development of improved detection method based on molecular approaches. In the gene detection assay, Lipopolysaccharide gene of Salmonella, V. cholera and E. coil were hybridized to anti-LPS factor gene found in the biolysate of the marine animals. The wzm and wzt genes encoding O-polysaccharide genes were amplified in these pathogens and the LPS factor C were amplified from the marine lysate. Development of a PCR-based technique for detection of the food-borne pathogens particularly Sa Salmonella, V. cholera and E. coil were achieved. Thus rapid, sensitive and reliable techniques for the detection of food-borne pathogens developed.
  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.
  Nurul H. Idris , Nagi A. Al-Haj , Mariana N. Shamsudin and Raha A. Rahim
  Safe attenuation has been done on marine pathogen Vibrio alginolyticus using naturally acidified fructose against vibriosis. Attenuation was confirmed by injecting the attenuated bacterium into fish where the survival rate was 100% compared to 50% survival in fish injected with non-attenuated bacteria. The attenuated bacterium was then evaluated for oral vaccination of Lates calcarifer (Asian seabass). Fish were fed with fish pellet incorporated with attenuated and non-attenuated bacterium of V. alginolyticus for 30 days. They were measured for serum antibody production by conventional agglutination titer and also monitored for the fish weight gain to observe the health improvement. Vaccinated fish showed comparable increased in weight gain, 90% survival after challenge and significantly high antibody titer compared to other treatment and control.
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