Despite health literacy being an important assumption for maintaining a healthy population, there are not many studies focusing on health literacy concerning HIV infection among adolescents in the Czech Republic and among Czech authors. In contrast, there are plenty of scientific papers and researches focused on awareness and susceptibility of adolescents in most affected regions of the world, Sub-Saharan Africa and India particularly. Adolescents belong to a high-risk group of the population. Education is one of the most important factors that influence the level of risk of HIV infection. However, increasing health literacy is only implemented in some types of secondary schools in the Czech Republic. The goal of this study is to verify health literacy concerning HIV infection among adolescents in two types of Czech secondary schools (grammar school and vocational school) in two Czech regional cities (Plzeň and České Budějovice). To determine the level of health literacy of HIV infection, a questionnaire (13 open and 9 closed questions) was compiled. A descriptive cross-sectional study was implemented among students (N = 1 942) of 16 chosen schools in two cities of the Czech Republic in 2015. The questionnaire was pre-tested and it took about 15 minutes to complete. The association was tested by a chi-square test, the averages were then compared with an independent t-test, the p-value (0,05) was used in all tests. Most of the students (67,3%) had good health literacy about HIV/AIDS, statistically significant differences were found between students of grammar schools and vocational schools (p = 0,012). Differences were also found between men and women (p = 0,037). The respondents showed the best knowledge about transmission (94,3%), the poorest about epidemiological questions (18,3%). Gender and the type of school had a statistically significant influence on the level of knowledge (gender x2; p = 0,042, types of a school x2; p = 0,031). In this cross-sectional study, women studying at grammar schools had the best knowledge of HIV/AIDS. In contrast, the lowest level of knowledge was found in men studying in vocational schools. The school types and gender had a statistically significant influence on the level of knowledge. However - knowledge is not enough. To reduce infection risk potential will mean to help adolescents to transform knowledge into diminishing of their risky behaviour. The implementation of prevention in all types of schools is crucial for the reduction of exposure from HIV infection. We consider educating adolescents about sexually transmitted diseases important. Despite that our research showed differences among schools and gender. Those differences can make a particular group be more at risk of being infected. Based on the findings, it would be suitable to unify and improve the approach to the education of this topic in all types of schools and to target preventive and educational techniques to boys more, since men are the biggest group of the HIV positive people in the Czech Republic.

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