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I Putu Bayu Mayura
Department of Clinical Microbiology, Medical School, Faculty of Medicine, Udayana University
Biochemistry, Genetics & Molecular Biology Environmental Science

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DETECTION METALLO-BETA-LACTAMASE GENE IMP-1 AND IMP-2 OF Pseudomonas aeruginosa CLINICAL ISOLATES IN SANGLAH HOSPITAL BALI Ni Made Adi Tarini; Ni Nengah Dwi Fatmawati; I Putu Bayu Mayura
International Journal of Biosciences and Biotechnology Vol 3 No 1 (2015)
Publisher : Central Laboratory for Genetic Resource and Molecular Biology, Faculty of Agriculture, Udayana University in cooperation with Asia-Oceania Bioscience and Biotechnology Consortium (AOBBC)

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Abstract

Pseudomonas aeruginosa is a pathogen frequently found as an agent of Hospital Acquired infections. This bacterium is very easy to be resistant to several types of antibiotics through various mechanisms. Carbapenem such as Imipenem and Meropenem is a potential option for the therapy of this bacterium, but unfortunately P. aeruginosa has ability in hydrolyzing these antibiotics through enzyme metallo-?-lactamases (MBLs). Recently, IMP and VIM, MBLs enzyme group are reported common from various countries, but no data is reported for these enzymes in Indonesia especially in Bali. In fact, the resistant data of P. aeruginosa against carbapenem group antibiotics such as meropenem and imipenem is quite high in Sanglah General Hospital in 2014 was 35% and 45% respectively. Therefore, the aim of this study was to detect IMP-1 and IMP-2 genes of MDR P. aeruginosa, which are phenotypically resistant to the antibiotic Imipenem and Meropenem disks based on CLSI standards in Clinical Microbiology Laboratory, Sanglah General Hospital, Denpasar, Bali. Eighty-six isolates were isolated from sputum (25 / 29.1%), wound (25 / 29.1%), urine (15 / 17.4%),endotracheal Tube (11 / 12.8), pus (6/7% ), blood (3 / 3.5%) and tissue (1 / 1.1%). In this study, all isolates were subjected to PCR for detection of IMP-1 and IMP-2. The result showed that 9 isolates were positive IMP-1 gene (10.5%), but there was no isolate positive for IMP-2 gene. The result was similar with that of the other countries, especially for the gene IMP-1. Detection and molecular characterization of MBL-producing P. aeruginosa strains are very important for infection control purposes. Currently, this study is still continued for detection of another MBL genes.
MULTIPLEX PCR FOR DETECTION OF CAPSULAR POLYSACCHARIDES TYPES OF Streptococcus pneumoniae CLINICAL ISOLATES IN BALI Ni Nengah Dwi Fatmawati; Ni Made Adi Tarini; I Putu Bayu Mayura
International Journal of Biosciences and Biotechnology Vol 2 No 2 (2015)
Publisher : Central Laboratory for Genetic Resource and Molecular Biology, Faculty of Agriculture, Udayana University in cooperation with Asia-Oceania Bioscience and Biotechnology Consortium (AOBBC)

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Abstract

Streptococcus pneumoniae is causative agent of non-invasive and Invasive Pneumococcal Diseases(IPD). One of the major virulence factors is capsular polysaccharides (CPS). The CPS is known as thepneumococcal vaccine component. Several types of S. pneumoniae CPS are dominant in Indonesiasuch as types 6, 23, 15, 33 and 12 in West Nusa Tenggara, type 7F in Jakarta, and types 6A/B dan15B/C in Central Java. No data is reported from Bali related to S.pneumoniae CPS typing. Therefore,the aim of this study was to determine CPS types of S. pneumoniae isolates in Clinical MicrobiologyLaboratory, Sanglah General Hospital, Denpasar, Bali by using Multiplex PCR. Twenty-one isolatesthat were isolated from blood (11/52.4%), sputum (5/23.8%), and other clinical specimens (5/23.8%)were included in this study. Identification of S. pneumoniae was based on optochin test and presenceof pneumolysin gene (ply). Uniplex PCR was conducted to determine capsular type of each isolates,and then continued with Multiplex PCR 1 and 2, which used in-house positive controls. All isolateswere positive for the presence of ply, confirming the isolates were S. pneumoniae. Moreover, thisstudy showed that type 19F was the predominant type (7 isolates (66.7%)); 2 isolates (9.5%) werepositive for each type 23F and also for type 6A/B; and, there was only 1 isolate (4.8%) for each type7F and 15B/C. Total of 8 isolates (38.1%) were found to be nontypeable isolates. Multiplex PCR wassuccessfully identified different types of CPS. Development of Multiplex PCR could help indiagnosing and identifying capsular type of S. pneumoniae simultaneously.
Co-Authors Ni Made Adi Tarini Ni Nengah Dwi Fatmawati