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    A CASE OF FATAL DISSEMINATED INFECTION CAUSED BY MYCOBACTERIUM BOVIS BCG STRAIN AND THE IDENTIFICATION OF THE ISOLATE BY SPOLIGOTYPING
    (Ankara Microbiology Soc, 2010) Aslan, Gonul; Kuyucu, Necdet; Aydin, Esin; Gunal, Selami; Emekdas, Gurol
    The vaccine strain Mycobacterium bovis BCG may lead to disseminated infection in patients with immune deficiency In this report a patient who developed fatal disseminated tuberculosis caused by M bovis BCG strain was presented One year old male patient with the previous history of recurrent lower respiratory tract infection, was admitted to the hospital with the complaints of fever, cough and diarrhea continuing for 20 days There was no family history of tuberculosis or history of contact with a tuberculosis case. Physical examination of the case revealed growth retardation and reticular and reticulonodular infiltration was detected in his chest X-ray. The results of sweat test, cystic fibrosis gene mutation analysis and metabolic screening tests were normal. Since fever continued and infiltrations persisted in the chest X-ray despite antibiotic therapy, PPD test was applied and acid-fast bacilli (AFB) were investigated in his gastric aspirate and stool samples for three consecutive days. PPD test was negative and no AFB were detected in the microscopic examination of the clinical samples. However, growth in Lowenstein-Jensen medium was detected in the stool sample on the 38(th) day of incubation. The antimycobacterial susceptibility testing performed at BACTEC MGIT (Mycobacterial Growth Indicator Tube) 960 system (Becton-Dickinson, USA) revealed that the isolate was susceptible to rifampin, isoniazid, streptomicin and ethambutol Since the isolates did not grow at PNB (para-nitro benzoic acid) medium and niacin and nitrate activities were negative, spoligotyping (spacer oligonucleotide typing) was performed and DR loci characteristic for M bovis BCG strain were detected. However, the patient died 2 weeks before the culture results were obtained The effective use of mycobacteriology laboratories and cooperation between laboratory and clinics provide advantages in the early diagnosis and treatment of tuberculosis cases, decreasing the morbidity and the mortality.
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    Isolation and Identification of Mycobacterium bovis and Non-tuberculous Mycobacteria in Raw Milk Samples in Mersin Province
    (Ankara Microbiology Soc, 2012) Aydin, Fatma Esin; Ulger, Mahmut; Emekdas, Gurol; Aslan, Gonul; Gunal, Selami
    This study was aimed to isolate and identify Mycobacterium bovis and non-tuberculous mycobacteria (NTM) species in raw milk samples from cattles and to compare the diagnostic performance of the methods used for that purpose. A total of 145 raw milk samples from cattles were collected from five villages in Mersin province (located on Mediterrenean region of Turkey) between April and June 2008. Presence of mycobacteria was investigated by Ehrlich Ziehl Neelsen (EZN) staining method, culture in Lowenstein-Jensen (LJ) medium and polymerase chain reaction (PCR). Only 1 (0.7%, 1/145) raw milk sample was found to be acid fast bacilli (AFB) positive with EZN staining. Eleven (7.6%) samples were positive by culture and mycobacterial DNA was detected in 6 (4.1%) samples by PCR. Mycobacterium was isolated from both creamy and pellet layer of a culture positive sample. Identification was carried out with conventional biochemical tests, PCR-Restriction Fragment Length Polymorphisms (PCR-RFLP) and spoligotyping (spacer oligonucleotide typing) methods. One isolate was identified as Mycobacterium tuberculosis complex (MTC) and 11 isolates were identified as NTM out of 12 isolates those were isolated from culture. According to PCR-RFLP analysis of these 11 NTM isolates, 6 (54.5%) were Mycobacterium genavense, 2 (18.2%) were Mycobacterium simiae, 2 (18.2%) were Mycobacterium szulgai and 1 (9.1%) was Mycobacterium fortuitum. MTC isolate was identified as M.bovis by spoligotyping. According to the results of our study, both pellet and creamy layers from raw milk samples should be cultured to selective LJ medium (without glycerol, with 0.4% sodium pyruvate) to improve the chance of isolation and must be incubated for up to eight weeks. In our region, NTM were isolated in 6.9% and M.bovis in 0.7% of the raw milk samples from cattles and this emphasized the risk of transmission of mycobacteria to man via direct contact or ingestion of unpasteurized milk products.
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    Isolation and identification of Mycobacterium tuberculosis from the urine samples by conventional and molecular methods
    (Ankara Microbiology Soc, 2007) Aslan, Gonul; Doruk, Erdal; Emekdas, Gurol; Serin, M. Sami; Direkel, Sahin; Bayram, Gul; Durmaz, Riza
    Genitourinary tuberculosis presents a challenge in diagnosis and treatment due to variations in clinical and radiological signs, insufficient patient history and difficulty in the isolation of the bacilli. The aim of this study was to isolate and identify Mycobacterium tuberculosis from the urine samples obtained from patients with suspected urinary tuberculosis admitted to our hospital by using Ehrlich-Ziehl-Neelsen (EZN), culture and polymerase chain reaction-restriction analysis (PCR-RFLP) methods. A total of 1004 urine samples collected from 437 patients who were admitted to our hospital between January 2004- July 2006, were inoculated on Lowenstein-Jensen (LJ) and/or BACTEC 12B (Becton Dickinson, USA) after decontamination and, direct preparations stained with EZN method were evaluated microscopically. M.tuberculosis complex (MTC) and mycobacteria other than tuberculosis (MOTT) were differentiated by nitro-alpha-acetylamino-beta-hydroxypropiophenone (NAP) test and the susceptibility testing for the MTC strains to primary antituberculosis drugs were performed by BACTEC 460 TB (Becton Dickinson, USA) system. PCR-RFLP method was performed for the identification of Mycobacterium spp. Twenty-two (5%) patients have yielded positive results by at least one of the conventional methods (EZN, LJ and/or BACTEC). Fifteen samples were positive for acido-resistant bacilli (ARB) by EZN method, and 17 samples were positive for mycobacterial growth in the cultures. Ten of 22 patients were found positive by both of the methods, while seven were culture positive but ARB negative and five were culture negative but ARB positive. These five patients received BCG treatment because of the presence of bladder tumor. Twelve (70.5%) of 17 strains isolated from culture were identified as MTC, while five (29.4%) were identified as M.fortuitum. Of 12 MTC isolates, eight (66.7%) were found susceptible to all of the antituberculosis agents, while one was found resistant to isoniazide (INH) and ethambutole (ETB), one was resistant to INH and rifampicin (RIF), and two were resistant to only INK It is concluded that, in order to identify mycobacteria and to perform antituberculous susceptibility tests, cultivation of mycobacteria is a prerequisite.