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Title: Phenotypic and genotypic typing of Burkholderia cepacia complex isolated from non-cystic fibrosis patients in two governmental secondary hospitals in Kuwait
Authors: Dana Wai'l Jawad Al-Kayyali 
Supervisor: Dr. Leila Vali
Keywords: complex isolated , non-cystic fibrosis patients
Issue Date: 2018
Publisher:  Kuwait university - college of graduate studies
Abstract: Burkholderia cepacia complex (Bcc) is a group of opportunistic pathogens comprising of at least 20 closely related species that are phenotypically similar yet genetically distinct. Bcc is prevalent in nature and manmade environments due to its metabolic capacity. The focus on Bcc species in this study is driven by their pathological significance and the lack of precise reports concerning their prevalence in Kuwait. In this study we aimed to perform phenotypic and molecular characterization of multidrug resistant Bcc isolates from Mubarak Al-Kabeer Hospital (MKH) and Amiri Hospital in Kuwait. Eight isolates within six months were collected from patients residing in Mubarak Al-Kabeer Hospital and Amiri hospital with no history of cystic fibrosis from different sites of infection. Phenotypic identification was performed using VITEK® 2 compact system and API20NE kit. Isolates were tested for antimicrobial susceptibility using E-Test strips and agar dilution method to determine the MICs. Genotypic identification methods including PCR for recA gene, PCR-RFLP for recA, Pulsed-field Gel Electrophoresis (PFGE) and Multi-locus sequence typing (MLST) were applied. All samples were identified as Bcc. A PCR product of 1 kb Bcc specific recA gene was amplified from all samples. Three different patterns were observed by PCR-RFLP for recA including B. cenocepacia (genomovar IIIA) [n=6], B. cepacia (genomovar I) [n=2] and B. multivorans (genomovar II) [n=1]. Results of PFGE using SpeI restriction endonuclease illustrated the genomic diversity of the isolates. We report for the first time three novel STs (ST-1282, ST-1284, ST-1288) and three novel allelic profiles for gltB (563), lepA (523) and gyrB (846) for the newly identified ST 1288. Interpretation of MIC values showed all isolates were resistant to at least one group of antibiotics: ceftazidime (n=1), meropenem (n=2), minocycline (n=1), levofloxacin (n=1), and chloramphenicol (n=8). One of the isolates was found to be resistant to trimethoprim-sulfamethoxazole. ST306 B. cenocepacia IIIA was the most prevalent type isolated from blood, of which; one isolate was pan-resistant and ultimately resulted in sepsis and death of a patient. This is the first study performed to characterize the Bcc strains in Kuwait. The results will contribute to better understanding of genetic relatedness of isolates from Kuwait to other regions of the world.
Appears in Programs:0712 Medical Laboratory Sciences

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