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- and 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- and 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.

National Laboratory Surveillance of Influenza in Macedonia during Four Seasons.

OBJECTIVE: The aim of the study is to comprehend results of the influenza lab surveillance system in the Republic of Macedonia after the 2009 pandemic and to determine the main characteristics of four consecutive epidemic seasons (from 2010/2011 until 2013/2014). METHODS: As part of the universal surveillance system, nasal and throat specimens were collected from patients. After extraction of RNA, the CDC real-time RT-PCR assays for the detection of influenza types and subtypes were performed. RESULTS: Out of 920 tested samples, 406 (44.1%) laboratory confirmed cases of influenza were found. Influenza activity begins as early as December and continues until the end of April with peaks in January or February with predominant influenza A and A/H1N1pdm. Influenza A viruses start their activity at week 49 to 52 and subside at week 17. Usually two peaks appear, the first one between week 2 and 4 and the second one between week 6 and 9. Subtype A/H1N1pdm was dominant among influenza A types in the 2010/2011 and 2012/2013 seasons. A/H3N2 was the only circulating influenza virus in the 2011/2012 season. Influenza B season is shorter and has only one peak, between weeks 2-5. Usually the influenza B viruses emerge in later stages than influenza A viruses, except for the first post-pandemic season. CONCLUSION: Results revealed that post-pandemic influenza seasons in Macedonia were rather different. Although the influenza season pattern is similar to patterns in some countries of the WHO European region, some unique characteristics were observed.

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 µg/ml. Three vanB genotypes of E. faecium and one vanB genotype of E. faecalis had MICs of 32 µg/ml. All six vanC genotypes of E. gallinarum had MICs of 8 µg/ml. All vanA genotypes of VRE were highly resistant to vancomycin, with MICs above 256 µg/ml. Three vanB genotypes of VR E. faecium and one VR E. fecalis were resistant, with MICs 32 µg/ml. vanC genotypes of VR E. gallinarum were intermediate resistant to vancomycin with MICs of 8 µg/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 µg/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.

Comparison of Directigen Flu A+B with Real Time PCR in the Diagnosis of Influenza.

UNLABELLED: Early diagnosis and treatment of patients with influenza is the reason why physicians need rapid high-sensitivity influenza diagnostic tests that require no complex lab equipment and can be performed and interpreted within 15 min. The Aim of this study was to compare the rapid Directigen Flu A+B test with real time PCR for detection of influenza viruses in the Republic of Macedonia. MATERIALS AND METHODS: One-hundred-eight respiratory samples (combined nose and throat swabs) were routinely collected for detection of influenza virus during influenza seasons. Forty-one patients were pediatric cases and 59 were adult. Their mean age was 23 years. The patients were allocated into 6 age groups: 0-4 yrs, 5-9 yrs, 10-14 yrs, 15-19 yrs, 20-64 yrs and > 65 yrs. Each sample was tested with Directigen Flu A+B and CDC real time PCR kit for detection and typisation/subtypisation of influenza according to the lab diagnostic protocol. RESULTS: Directigen Flu A+B identified influenza A virus in 20 (18.5%) samples and influenza B virus in two 2 (1.9%) samples. The high specificity (100%) and PPV of Directigen Flu A+B we found in our study shows that the positive results do not need to be confirmed. The overall sensitivity of Directigen Flu A+B is 35.1% for influenza A virus and 33.0% for influenza B virus. The sensitivity for influenza A is higher among children hospitalized (45.0%) and outpatients (40.0%) versus adults. CONCLUSION: Directigen Flu A+B has relatively low sensitivity for detection of influenza viruses in combined nose and throat swabs. Negative results must be confirmed.

Methods for molecular surveillance of influenza used in Macedonia.

THE AIM: To present and compare different Nucleic Acid Testing assays used for laboratory diagnosis of influenza virus infection in our country. MATERIALS AND METHODS: Respiratory samples used were nose and throat swabs. The RNA extraction was performed with a QIAamp viral RNA kit. During the season 2009­2010 the first 25 samples were tested with: conventional gel-based RT-PCR and CDC rtRT-PCR using published specific matrix and HA gene primers and probes for influenza virus typing and subtyping. RESULTS: Of 25 samples tested with conventional RT-PCR 7(28%) were positive for influenza A, but negative for A/H1seasonal and A/H3. Retested with rtRT-PCR 9(36%) were positive for influenza A, 8(32%) were positive for А/H1pdm and 1(4%) was А/H3. Two samples positive with rtRT-PCR for influenza A were negative with RT-PCR. The sensitivity of the RT-PCR in comparison with rtRT-PCR is 100% and the specificity is 88.89%. Positive predictive value for RT-PCR is 77.78%, and negative predictive value is 100%. RT-PCR is a four-step and rtRT-PCR a one-step procedure. The turn-around time of RT-PCR is 6 hours and for rtRT-PCR it is 2 hours. DISCUSSION AND CONCLUSION: For surveillance purposes nose and throat swabs are the more easy and practical to collect. It was proved that RT-PCR is too laborious, multi-step and time-consuming. The sensitivity of both assays is equal. The specificity of rtRT-PCR is higher. NAT assays for detection of influenza viruses have become an integral component of the surveillance programme in our country. They provide a fast, accurate and sensitive detection of influenza.

Laboratory surveillance during pandemic influenza season 2009/2010 in Republic of Macedonia.

OBJECTIVE: To provide virological and epidemiological information on patients laboratory-tested for influenza A/ H1N1pdm during the pandemic season April 2009/May 2010. MATERIALS AND METHODS: Demographic and other data were obtained from the request form arriving with the samples of patients whose symptoms met the clinical definition of influenza A infection. The RNA was tested for the presence of influenza virus using the CDC real-time RT-PCR assay. A total of 3010 suspect patients (pts) were tested from week 18 2009 to week 20 2010. RESULTS: 1632 pts (54.2%) were oositive for influenza. From them 1556 samples were confirmed as H1N1pdm. There was a domination of H1N1pdm positivity among young persons in age groups 5-17 (34.4%) and 18-49 (31.4%) years. The pandemic influenza was presented in two waves. The first wave started on 20 June with the first positive case and peaked early in August (week 32). The second wave started from week 44. The majority of positive cases were between week 45 and week 52. 37.7% of the positive pts were hospitalized--66.7% of pts at age 65+ and 63.3% of children in the age group 0-4 years. The highest percentage of patients with underlying medical conditions were in the age group 50-64 (49.35%) years and 65+ (88.23%) years. 1.15% of the positive pts for H1N1pdm gave data for vaccination with seasonal influenza. CONCLUSIONS: Data obtained from laboratory and epidemiological surveillance of pandemic influenza will serve public health to a full understanding of the pandemic 2009/2010 influenza in R. Macedonia and dealing with future challenges.

The first survey about the antibiotic usage in the Republic of Macedonia.

Development of antibiotic resistance represents a major global and Macedonian public health problem. To assess the opinion and knowledge of the citizens of Macedonia about the usage of antibiotics, voluntary and anonymous survey was realized. A total of 239 persons (age 10-67 years) were interviewed. Following information was obtained: 73.64% get antibiotics with a medical prescription; and 87.03% receiving the antibiotic on time, dosage and prescribed duration. When asked about knowledge about antibiotics, 38% of the interviewed persons gave the right answer, 43.1% of respondents made false statements that antibiotics are effective against viral infections and 25.52% did not express any opinion.

Chlamydia pneumoniae and helicobacter pylori serology - importance in patients with coronary heart disease.

BACKGROUND: Chronic infections in CHD are due to one or both of the organisms Chlamydia pneumoniae and Helicobacter pylori. AIM: To examine the association between serum markers of Chlamydia pneumoniae and Helicobacter pylori infection and markers of myocardial damage. in patients with acute coronary syndrome (ACS), with chronic coronary artery disease (CAD) and in-control group. MATERIAL AND METHODS: Sera were taken from a total of 153 subjects. Subjects were divided in three groups: 64 patients with ACS; 53 patients with CAD and a group of 35 conditionally healthy individuals. Analysis of patients' sera for IgG antibodies to H. pylori and markers for myocardial damage was done on the Immulite system. The presence of specific IgG and IgA antibodies to C. pneumoniae was determined with MIF, Sero FIA (Savyon Diagnostics, Israel). Statistical analysis of data was done using the statistical program SPSS (Statistical Package for Social Sciences), version 13. RESULTS AND DISCUSSION: There was a high significant difference in troponin levels between the three groups of subjects (p=0.0000). Levels of creatine kinase isoenzyme (CK-MB) were highest in the ACS group (500.0 ng/mL). There was a statistically significant difference between CG subjects and ACS patients due to more frequent detection of antichlamydial IgA antibodies in patients with acute coronary syndrome. Positive serum immune response for Helicobacter pylori was 17 (53.1%) and 29 (80.6%), respectively. CONCLUSION: Increased IgA antibody titers for C. Pneumoniae, increased CRP values as well as classic markers of myocardial damage are risk factors for coronary events.

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 masculineC, RT-PCR reaction was identical to the one performed initially. The obtained amplicons were maintained frozen at -20 masculineC 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 specificity.

Initial empiric antimicrobial treatment of Chlamydia pneumoniae: a study of 54 cases in the Republic of Macedonia.

UNLABELLED: Analysis of the adequacy of the initial empiric antimicrobial treatment of patients with acute Chlamydia pneumoniae (CP) infection, admitted to the Infectious Diseases Clinic, Clinical Centre, Medical Faculty, Skopje, Republic of Macedonia, for community-acquired pneumonia. MATERIAL AND METHODS: A total number of 407 patients with community-acquired pneumonia (CAP) hospitalized at the Clinic between September 1997 and June 2002, with an average age of x=46.44 years, of whom 53.56% were male. Acute Chlamydia pneumoniae infection was proven serologically with MIF assay in 54 (13.27%) patients. RESULTS: Initial empiric treatment with antibiotics in patients with CP pneumonia was provided with antimicrobial agents with intracellular activity in 26 (48.15%) patients; with fluoroquinolones in 19 (35.19%); macrolides in 5 (9.26%) and tetracyclines in 2 (3.7%). The treatment was conducted as monotherapy in 6 patients (11.11%) and in 20 patients (37.04%) in combination with betalactams. For 28 (51.85%) patients who were treated only with betalactams, empiric treatment was re-evaluated and new therapy with fluoroquinolon was conducted in 16 (29.63%), with macrolides in 8 (14.81%) and with tetracyclines in 4 (7.41%) patients. CONCLUSION: Adequate empiric treatment with antimicrobial agents with intracellular activity was performed in only 48.15% of the patients with acute CP infection. Therefore, when designing the initial empiric treatment of patients hospitalized with pneumonia, attention should be paid to this atypical pathogen of CAP.

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 microg 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.