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.

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.

Nutritional status and growth parameters of school-age Roma children in the Republic of Macedonia.

Main objective of the study was to assess the nutritional status of school age Roma children in Macedonia in order to detect precursors of possible health risks at an early age. The study was designed as a comparative case control study. Study group consisted of 229 Roma school children from the 1st and 272 from the 5th grade residing in different towns in Macedonia. The control group was recruited from other than Roma ethnic background and consisted of 283 children attending 1st and 356 children attending 5th grade. Every participant was measured for his/hers body height and weight. The t-test and Chi square (Chi2) were applied to test statistical significance of variables. The WHO's AnthroPlus software was applied to assess growth parameters and population at risk. There were significant differences in values of the body weight (p = 0.001) and height (p = 0.001) between Roma and non-Roma children attending the 1st grade of primary school. Weight-for-age, height-for-age and BMI-for-age indexes of the 1st grade children significantly differred in in the same intervals of SD (> or = -2SD and < -1SD; > or = -1SD and median; > +1SD and < or = +2SD; between Roma and non-Roma 5th graders. Anthropometric parameters of nutritional status of Roma children in Macedonia are significantly different than those of their non-Roma peers. Their health risks are predominantly related to underweight. The parameters related to health risks of overweight or obesity are lower in Roma than in non-Roma children.