Gastrointestinal Colonization with Vancomycin-Resistant Enterococci in Hospitalized and Outpatients

BACKGROUND: The incidence of infection and intestinal colonization with vancomycin resistant enterococci (VRE) is increasing in many countries in the last decade. Concerning the difficult antimicrobial treatment of infections caused by VRE, decreasing the incidence and prevalence of these infections is an important factor in VRE-induced morbidity and mortality control.AIM: To determine the prevalence of gastrointestinal colonization with vancomycin resistant enterococci in hospitalized and outpatients, and to determine the genetic base of the vancomycin resistance in VRE isolates.MATERIAL AND METHODS: Seven hundred and eighty stool specimens were investigated for the gastrointestinal carriage of vancomycin-resistant enterococci (VRE). Susceptibility to vancomycin was tested in all isolates by disk-diffusion test and E-test (AB Biodisk, Sweden). Determined vancomycin resistant enterococci were than tested for detection of vanA, vanB and vanC genes by PCR.RESULTS: Vancomycin resistant strains of enterococci were isolated from 46 (16.1 %) of the 285 hospitalized patients and 5 (7.7 %) of the 65 patients living in the community (p < 0.05). The most of the highly resistant enterococci strains to vancomycin (95.2 %), were identified as E. faecium. Minimal inhibitory concentrations (MICs) to vancomycin in all 39 vanA genotypes of E. faecium and two vanA genotypes of E. fecalis were > 256 mg/ml. Three vanB genotypes of E. faecium and one vanB genotype of E. faecalis had MICs of 32 mg/ml. All six vanC genotypes of E. gallinarum had MICs of 8 mg/ml. All vanA genotypes of VRE were highly resistant to vancomycin, with MICs above 256 mg/ml. Three vanB genotypes of VR E. faecium and one VR E. fecalis were resistant, with MICs 32 mg/ml. vanC genotypes of VR E. gallinarum were intermediate resistant to vancomycin with MICs of 8 mg/ml.CONCLUSIONS: The prevalence of vancomycin resistant enterococci in Republic of Macedonia was 2-fold higher in hospitalized than in outpatients. VanA genotype was dominant in isolates of E. faecium and it was highly associated with the MIC values above the 256 mg/ml. Since most of the enterococcal infections are endogenous, there is a need for screening the colonization of patient’s intestinal flora with VRE at the hospital entry. Identification and genotyping of faecal enterococci, together with their susceptibility testing to vancomycin, could be useful marker for the infection control

Интрахоспитални инфекции - мултифакторски пристап во превенција на нозокомијалните инфекции

Домашната стручна литерарура е инсуфициентна во однос на познавањето на стратегиите за превенцијата со мултидисциплинарен приод, постоење на организиран пристап и прирачници со мерки за контрола на нозокомијалните инфекции. Целта на овој текст е да биде корисна литература за секој тим за контрола на интрахоспитални инфекции, кои според законските норми на Република Македонија е задолжен да ја организира контролата на нозокомијалните инфекции во својата здравствена институција. Содржината на книгата нуди упатства за воспоставување на добро организиран систем за контрола на инфекциите.Од тој аспект, може да послужи и како прирачник на сите здравствени работници кои се одговорни за контролата на хигиено-техничкиот и асептичниот процес во институцијата

RT-PCR Detection of HIV in Republic of Macedonia

The aim of the study was to detect HIV RNA in seropositive patients using RT-PCR method and thus, to establish PCR methodology in the routine laboratory works. The total of 33 examined persons were divided in two groups: 1) 13 persons seropositive for HIV; and 2) 20 healthy persons - randomly selected blood donors that made the case control group. The subjects age was between 25 and 52 years (average 38,5). ELFA test for combined detection of HIV p24 antigen and anti HIV-1 + 2 IgG and ELISA test for detection of antibodies against HIV-1 and HIV-2, were performed for each examined person. RNA from the whole blood was extracted using a commercial kit based on salt precipitation. Detection of HIV RNA was performed using RT-PCR kit. Following nested PCR, the product was separated by electrophoresis in 1,5 % agarose gel. The result was scored positive if the band of 210bp was visible regardless of intensity Measures of precaution were taken during all the steps of the work and HIV infected materials were disposed of accordingly. In the group of blood donors ELFA, ELISA and RT-PCR were negative. Assuming that prevalence of HIV infection is zero, the clinical specificity of RT-PCR is 100 %. The analytical specificity of RT-PCR method was tested against Hepatitis C and B, Human Papiloma Virus, Cytomegalovirus, Herpes Simplex Virus, Rubella Virus, Mycobacterium tuberculosis, Chlamydia trachomatis. None of these templates yielded amplicon. In the group of 13 seropositive persons, 33 samples were analyzed. HIV RNA was detected in 15 samples. ELISA and ELFA test were positive in all samples. Different aliquots of the samples were tested independently and showed the same results. After different periods of storing the RNA samples at -70°C, RT-PCR reaction was identical to the one performed initially. The obtained amplicons were maintained frozen at -20°C for a week and the subsequently performed electrophoresis was identical to the previous one. The reaction is fast, simple for manipulation; with low detection level of 60 IU/ml. RT-PCR needs a small amount of RNA, as well as a small volume of sample. HIV RNA was detected in different periods of time with different clinical presentations in patients, with or without antiretroviral therapy. RT-PCR method gives the opportunity for reliable determination of HIV-1 RNA with border of detection of 60 IU/ml. The test is reproducible and has high analytical and clinical specificit

RT-PCR Detection of HIV in Republic of Macedonia

The aim of the study was to detect HIV RNA in seropositive patients using RT-PCR method and thus, to establish PCR methodology in the routine laboratory works. The total of 33 examined persons were divided in two groups: 1) 13 persons seropositive for HIV; and 2) 20 healthy persons - randomly selected blood donors that made the case control group. The subjects age was between 25 and 52 years (average 38,5). ELFA test for combined detection of HIV p24 antigen and anti HIV-1 + 2 IgG and ELISA test for detection of antibodies against HIV-1 and HIV-2, were performed for each examined person. RNA from the whole blood was extracted using a commercial kit based on salt precipitation. Detection of HIV RNA was performed using RT-PCR kit. Following nested PCR, the product was separated by electrophoresis in 1,5 % agarose gel. The result was scored positive if the band of 210bp was visible regardless of intensity Measures of precaution were taken during all the steps of the work and HIV infected materials were disposed of accordingly. In the group of blood donors ELFA, ELISA and RT-PCR were negative. Assuming that prevalence of HIV infection is zero, the clinical specificity of RT-PCR is 100 %. The analytical specificity of RT-PCR method was tested against Hepatitis C and B, Human Papiloma Virus, Cytomegalovirus, Herpes Simplex Virus, Rubella Virus, Mycobacterium tuberculosis, Chlamydia trachomatis. None of these templates yielded amplicon. In the group of 13 seropositive persons, 33 samples were analyzed. HIV RNA was detected in 15 samples. ELISA and ELFA test were positive in all samples. Different aliquots of the samples were tested independently and showed the same results. After different periods of storing the RNA samples at -70°C, RT-PCR reaction was identical to the one performed initially. The obtained amplicons were maintained frozen at -20°C for a week and the subsequently performed electrophoresis was identical to the previous one. The reaction is fast, simple for manipulation; with low detection level of 60 IU/ml. RT-PCR needs a small amount of RNA, as well as a small volume of sample. HIV RNA was detected in different periods of time with different clinical presentations in patients, with or without antiretroviral therapy. RT-PCR method gives the opportunity for reliable determination of HIV-1 RNA with border of detection of 60 IU/ml. The test is reproducible and has high analytical and clinical specificit

Incidence of Staphylococcus aureus Isolated from Patients Treated at the Clinical Center of Skopje, Macedonia, with Special Attention to MRSA

The distribution of 3497 Staphylococcus aureus strains according to methicillin resistance, specimens, departmental profession and antibiotic resistance patterns was analysed. The strains were cultured from the patients of the Clinical Center of Skopje, Macedonia, between 1 January 2002 and 31 December 2004. The majority of the isolates was obtained from suppurated wounds (28.5%), nares (21%), intratracheal tubes (13%) and blood cultures (11.8%). Overall 1100 (31.4%) of the isolates was methicillin-resistant with 1 µg oxacillin disc. Of these 35.5%, 30.5% and 10.4% were cultured from wounds, intratracheal tubes and blood samples, respectively. The prevalence of MRSA strains was 78.6%, 75%, 44.2% and 37.3% in specimens of ICU, Coma Center, General Surgery and Haematology patients. There were extremely big differences in the frequency of MRSA between departments with particular specialisation. The 2397 MSSA isolates belonged to practically one antibiotic resistance pattern characterised with penicillin resistance and susceptibility to other antistaphylococcal drugs. The 1100 MRSA isolates distributed to four antibiotic resistance patterns on the basis of their resistance to oxacillin, penicillin, amoxicillin+clavulanic acid, azithromycin, clindamycin, amikacin, gentamicin, ciprofloxacin, trimethoprim+sulphamethoxasole, vancomycin and teicoplanin. All the MRSA isolates were multidrug resistant but sensitive to glycopeptides

Clinical Investigations. Comparison of Directigen Flu A+B with Real Time PCR in the Diagnosis of Influenza / Сравнение Иммунохроматографического Метода (Directigen Flu A+B) И Теста RT-PCR При Инфекциях Гриппа

Для ранней диагностики и лечения пациентов с гриппом нужны высокочувствительные диагностические тесты, применение которых не требует сложного лабораторного оборудования, а продолжительность проведения и читки реакции не превышает 15 минут. ЦЕЛЬЮ данного исследования является сопоставительное сравнение быстрого теста Directigen Flu A+B с тестом PCR в режиме реального времени при выявлении вирусов гриппа в Республике Македонии. МАТЕРИАЛЫ И МЕТОДЫ: В течение нескольких месяцев распространения сезонного гриппа было проведено 108 дыхательных проб (комбинированные мазки из носа и горла). Среди пациентов был 41 ребёнок и 59 взрослых, средний возраст которых составлял 23 года. Пациенты были разделены на следующие возрастные группы: 0 - 4, 5 - 9, 10 - 14, 15 - 19, 20 - 64 лет и старше 65 лет. Каждая проба была протестирована с использованием Directigen Flu A+B и тестового набора в режиме реального времени PCR CDC на выявление и определение типа и подтипа вируса гриппа в соответствии с лабораторным диагностическим протоколом. РЕЗУЛЬТАТЫ: Тест Directigen gen Flu А+В выявил 20 (18.5%) проб с положительным результатом на вирус гриппа А и 2 пробы (1.9%) с положительным результатом на вирус гриппа В. Высокая степень специфичности (100 %) и высокая степень положительного прогнозируемого значения, которые продемонстрировал тест Directigen Flu A+B во время нашего исследования являются показателем того, что положительные результаты не нуждаются в подтверждении. Общая чувствительность теста Directigen Flu A+B составляет 35.1 % на вирус гриппа типа А и 33.0 % на вирус гриппа типа В. Тест на вирус гриппа типа А проявил более высокую чувствительность при детях (госпитализировано 45.0 %, амбулаторных - 40.0 %) по сравнению с аналогичной при взрослых. ЗАКЛЮЧЕНИЕ: Тест Directigen Flu A+B обладает относительно низкой чувствительностью при выявлении вирусной инфекции в мазках из горла и носа. Отрицательные результаты подлежат верификации

Медицинска микробиологија - Општ дел

Во прилог скениран Предговор

Building antimicrobial stewardship through massive open online courses : A pilot study in Macedonia

Background: The global struggle against antibiotic resistance requires antimicrobial stewardship (AMS). Massive open online courses (MOOCs) offer health professionals unprecedented access to high-quality instructional material on AMS; the question is how apprehensible it is to non-native English speakers. Furthermore, to better understand how education interventions promote change towards rational antibiotic prescribing, leading institutions call for studies integrating behavioural science. Research from lower- A nd middle-income countries is particularly needed. Objectives: To measure the knowledge improvement from an AMS MOOC, the influence of language, course satisfaction and subsequent effect on intention to change antibiotic prescribing behaviour. Methods: Fifty-five physicians from Macedonia completed the MOOC. Pre- A nd post-course knowledge test scores were compared using a one-sample t-test. The effect of a language barrier was assessed using self-reported English level. Scores were compared with participants' intention to change behaviour in clinical practice. Results: Scores significantly improved from 77.8% to 82.2%. Participants with a higher English level improved most, while the low-level group showed no significant improvement. Physicians reported a high or very high intention to change behaviour. This was independent of knowledge improvements. Conclusions: First, lower self-reported English proficiency hindered knowledge acquisition from a MOOC platform. AMS programmes should commit to bridge this barrier so as to enable a global spread of education in AMS. Second, factors underlying the physicians' intentions to engage in AMS appear to be more complex than simple knowledge improvements. This suggests that less time-consuming interventions could be as effective

Formulation of synbiotic soy-based food product with antihypertensive potential

The specific aims of this study were to select the favorable prebiotic for L casei-01 as well as the suitable inoculum of the probiotic for fermented soy with ACE-inhibitory potential. For that purpose the metabolic activity of L casei-01 in soymilk supplemented with(1.5% w/v) prebiotics Fructooligosaccharide (FOS) or oligofructose enriched inulin (Synergy 1) was assessed. The evaluated parameters were: pH, viable cell counts, proteolysis, organic acid production and inhibition of angiotensine converting enzyme activity (IACE). The cell growth of L casei-01 reached the recommended therapeutic level of 9.58 ± 0.035 log cfu mL-1 for low inoculum samples (0.005 and 0.01%w/v) and 11.543 ± 0.13 log cfu mL-1 for high inoculum samples (0.075 and 0.1%w/v) regardless of the prebiotic used. The lower pH during fermentation, faster cell growth and superior proteolysis in Synergy 1 samples indicated better utilization of that prebiotic vs. FOS. The hydrolysis depended on the prebiotic used, showing higher values in Synergy 1 samples. The faster proteolysis was confirmed by SDSPAG electrophoresis. The Mw of polypeptides in the synbiotic end-products were lower than 30kD. The observed values for inhibition of ACE activity were app. 71, 74, 77 and 78% for inoculum rates of 0.005, 0.01, 0.075 and 0.1% w/v, respectively. Based on the results obtained in our study, the prebiotic Synergy 1 (1.5% w/v) and L. casei-01 at inoculum of 0.01% w/v for low dose and 0.075% w/v for high dose were considered more favorable for the production of synbiotic soy drink with antihypertensive potential

Synthesis, physicochemical characterization and antibacterial activity of novel (benzoylamino)methyl derivatives of quinolones

Herein we report the synthesis of different derivatives of (fluoro)quinolones norfloxacin, ciprofloxacin and pipemidic acid, by incorporating (benzoylamino)methyl on the free nitrogen of the pyperazinyl moiety. The compounds were structurally characterized by 1D and 2D NMR, FTIR and high resolution mass spectroscopy. In addition, their physicochemical properties were a matter of interest to be correlated with their structure and antimicrobial activity in vitro. Their antimicrobial activities were screened against Gram-positive, Gram-negative bacteria and C. albicans. Higher distribution coefficients and consequently lower water solubility were determined for all synthesized compounds than the ones of the corresponding leading compounds. Inconsequential correlations between the lipophilicity of the compounds and MIC were observed, suggesting that passive diffusion is not the only mechanism for their penetration into bacterial cells. Further studies are needed to determine how substitutions in the (fluoro)quinolone moiety affect the primary target(s), substrate behavior in respect to bacterial transporters and overall bioavailability