MATERIALS AND METHODS: The object of the study were samples of biological substrates (leukocyte mass, saliva, urine) taken from patients who underwent liver and kidney transplantation. Detection of CMV DNA was carried out by a real-time PCR using commercial diagnostic AmpliSense CMV-FL test systems (Central Research Institute for Epidemiology, Moscow, Russia). DNA extraction was performed using DNA-sorb AM and DNA-sorb V kits (Central Research Institute for Epidemiology) in accordance with manufacturer's manual. The quality of the prepared DNA library for sequencing was assessed by means of the QIAxcel Advanced System capillary gel electrophoresis system (QIAGEN, Germany). Alignment and assembly of nucleotide sequences were carried out using CLC Genomics Workbench 5.5 software (CLC bio, USA). The sequencing results were analyzed using BLAST of NCBI server.
RESULTS: CMV DNA samples were selected for genotyping. The two variable genes, UL55(gB) and UL73(gN), were used for CMV genotype determination, which was performed using NGS technology MiSeq sequencer (Illumina, USA). Based on the exploratory studies and analysis of literature sources, primers for genotyping on the UL55(gB) and UL73(gN) genes have been selected and the optimal conditions for the PCR reaction have been defined. The results of sequencing the UL55(gB) and UL73(gN) gene fragments of CMV clinical isolates from recipients of solid organs made it possible to determine the virus genotypes, among which gB2, gN4c, and gN4b were dominant. In some cases, association of two and three CMV genotypes has been revealed.
CONCLUSION: The application of the NGS technology for genotyping cytomegalovirus strains can become one of the main methods of CMV infection molecular epidemiology, as it allows for obtaining reliable results with a significant reduction in research time.
Methods: A cross-sectional, questionnaire-based study was conducted among 482 medical, dental, and pharmacy students of 3rd- and 4th-year degree program of Asian Institute of Medicine, Science and Technology University to assess their KAP about HB infection using 34 prevalidated questions by convenient sampling method. A questionnaire was administered to the students, and their responses were measured at "yes" and "no" scale. Students' responses were entered in SPSS version 22, and quantitative analysis was performed using descriptive statistics and nonparametric tests at P < 0.05.
Findings: The medical, dental, and pharmacy students had good knowledge and practice with positive attitude about HB infection. Mann-Whitney U-test determined a significant difference in knowledge (P < 0.001) and practice (P < 0.001) scores between medical and pharmacy, attitude (P < 0.001) scores between medical and dental, and attitude (P < 0.001) and practice (P < 0.001) scores between pharmacy and dental students. Students' age was correlated with their attitude, practice, and KAP scores and family income with their knowledge, attitude, practice, and KAP scores.
Conclusion: Although students' knowledge and practice were good with positive attitude, all the students did not participate in health education program, screening, and vaccination of hepatitis B virus (HBV) infection which makes them more vulnerable to occupational HBV infection. Hence, it is recommended to organize a regular health education program for the students on screening and vaccination against HBV to prevent its infection.