Ongoing measles outbreak, the Federation of Bosnia and Herzegovina, December 2023 to February 2024.

We report on an ongoing measles outbreak in the Federation of Bosnia and Herzegovina with 141 cases notified between week 52 2023 and week 6 2024. Among those with known vaccination status, 97% were unvaccinated and the most affected group is children under the age of 5 years (n = 87) who were not vaccinated during the pandemic years. Sixty-eight cases were hospitalised, the most common complications were measles-related pneumonia and diarrhoea. The sequenced measles viruses from four cases belonged to genotype D8.

Mpox virus DNA detection in different clinical specimens: tertiary-level laboratory experience, Bosnia and Herzegovina, 2022.

Mpox virus (MPXV) infection is difficult to distinguish from other (non-)infectious diseases. The etiology of rash can be differentiated by real-time polymerase chain reaction (rtPCR) on different types of samples. The study aims to provide experience with emerging MPXV diagnostics in a tertiary-level laboratory in Bosnia and Herzegovina. From July-December 2022, a total of 18 mpox suspected persons were tested. MPXV infection was confirmed by rtPCR in 10/18 (55.56 %) persons. The number of cases reached a peak in October 2022. The lowest median Crossing point (Cp) (x̄ = 29.76) was obtained from a swab of skin lesions in a viral transport medium (VTM). Evaluating the Cp values for the 7/9 mpox cases from whom paired swab samples from different anatomic sites were collected, higher positivity of skin lesion swabs in VTM was observed. In conclusion, our data highlighted the confirmatory role of rtPCR in the diagnosis of MPXV in skin lesion samples.

Expansion of hantavirus infection during the SARS-CoV-2 pandemic in Bosnia and Herzegovina, 2021.

Introduction. Bosnia and Herzegovina (B and H) has been recognized for decades as a country with a high risk of diseases caused by hantaviruses.Gap statement. The severe acute respiratory syndrome-associated coronavirus 2 (SARS-CoV-2) pandemic has diverted attention from many pathogens, including hantavirus.Aim. To provide a socio-demographic, temporal, geographical and clinical laboratory overview of the expansion of hantavirus infection cases during the SARS-CoV-2 pandemic in B and H in 2021.Methodology. The RecomLine HantaPlus IgG, IgM immuno-line assay (Mikrogen, Germany) was used to detect IgG and IgM antibodies to hantavirus serotypes in human sera from clinically suspected cases.Results. In 2021 (January-October), the number of confirmed cases of hantavirus infection and tested persons (92/140; 65,71 %) was higher than in the previous 2 years, 2020 (2/20; 10.00 %) and 2019 (10/61; 16.39 %). Most of the infected persons were men (84/92; 91.30 %). Hantavirus infections were recorded from January to October 2021, and the peak was reached in July (25/92; 27.17 %). Six out of 10 cantons in the Federation of Bosnia and Herzegovina (FB and H) were affected, namely Sarajevo Canton, Central Bosnia Canton, Neretva Canton, Zenica-Doboj Canton, Posavina Canton and Bosnian-Podrinje Canton Gorazde, in descending order. Of the 38/92 (41.30 %) infected patients with characteristic clinical manifestations of haemorrhagic fever, including renal (mainly) or pulmonary syndrome, 32/92 (34.78 %) were hospitalized in the Clinical Center of the University of Sarajevo. Two cases were detected with dual infection, hantavirus (Puumala) with Leptospira in one and SARS-CoV-2 in another case. The largest number of infections was related to Puumala (PUUV) (83/92; 90.22 %), while the rest of the infections were caused by the hantavirus Dobrava serotype (DOBV).Conclusion. The reported infections were probably caused by exposure of individuals to at-risk areas inhabited by contaminated rodents as natural reservoirs of hantavirus. As a highly endemic area, B and H requires continuous monitoring and increased awareness of this problem.

Performance Characteristics and Utility of the Standard Q COVID-19 Antigen Test for Emergency Admissions to Healthcare Facilities.

This study evaluated the performance of the COVID-19 Ag-RDT compared to the real-time reverse transcription-polymerase chain reaction (rtRT-PCR) for SARS-CoV-2 detection and its use among patients referred for emergency admission. A total of 120 nasopharyngeal swabs were collected from patients referred for emergency admission and immediately preceded for testing to the Unit of Clinical Microbiology. Out of 60 Ag positive tests, 53 (88.3%) were confirmed by rtRT-PCR, while 7 (11.7%) tested negative (false positives). Out of 60 Ag negative tests, 56 (93.3%) were confirmed negative by rtRT-PCR, and 4 (6.7%) were positive (false negatives). Ct value comparison was performed for 53 samples that were positive by both methods: 8 (15.1%) isolates had Ct value up to 20; 37 (69.8%) 21 to 30 and 8 (15.1%) 31 to 40, respectively. The sensitivity of the analyzed rapid Ag test was 92.9%, and specificity 88.9%. The accuracy of the Ag test was 90.8%. This study has shown that rapid Ag tests can be used in emergency admissions to healthcare facilities. However, rtRT-PCR should be considered after negative antigen test results in symptomatic patients, and after positive antigen test results in asymptomatic persons.

The SARS-CoV-2 Delta (B.1.617.2) variant with spike N501Y mutation in the shadow of Omicron emergence.

Circulation of the Omicron variant with the reemergence of the N501Y mutation along with many others in the spike protein has once again stirred the academic community. Interestingly, tracing the genetic diversity of SARS-CoV-2 shed light on a less frequent N501Y + Delta variant which has been in the global circulation for some time before the Omicron appearance. This paper aims to present the molecular characteristics of the SARS-CoV-2 Spike_N501Y + Delta variant detected in Bosnia and Herzegovina. The study was conducted during November and December 2021. All patients were tested using real-time RT-PCR for detection of SARS-CoV-2. A representative number of SARS-CoV-2 positive samples was pre-screened using VirSNiP SARS-CoV-2 Spike N501Y kit. The characterization of the viruses was carried out with Illumina RNA Prep with enrichment and the Respiratory Virus Oligo Panel kit. Among the analyzed sequences, we found two isolates of the Delta variant that differ from their most related clade- GK AY.4.3 in additional mutations N501Y and L54F. In this study, we described the presence of a rare form of Delta variant with Spike_N501Y mutation in the shadow of the Omicron emergence. Despite the set of mutations in the Spike protein, this form of Delta variant does not indicate the large-scale consequences for the general population. Further functional studies of this form could provide more information about its antigenicity and infectivity.

A novel flavivirus strain detected in phlebotomine sandflies in Bosnia and Herzegovina.

Aim Phlebotominae sandflies are primary vectors of phleboviruses, causing the sandfly fever disease. The aim of this study was to detect and report the presence of flaviviruses in Phlebotominae sandflies captured in Bosnia and Herzegovina. Methods After a microscopic and morphometric analysis, the final identification of collected Phlebotomus specimens was confirmed by PCR, using a hemi-nested polymerase chain reaction on extracted and reversely transcribed RNA. Results We obtained a 155 nt long fragment of the viral non-structural protein 5 (NS5) gene (GenBank accession no. MN090154). The acquired nucleotide sequence, provisionally named as Dreznica, showed a maximum of 70-80% identity in 70-88% (110-137 nucleotides) of the query coverage with several Anopheles, Sabethes, Calbertado and Culex flaviviruses. Maximum likelihood phylogenetic analysis showed that the new flavivirus Dreznica clusters together with the flavivirus isolated from Culiseta annulata mosquitos. Conclusion We report the presence of flavivirus in Phlebotominae sandflies, captured in Dreznica, Herzegovina for the first time. The next phase of research will be directed towards virus cultivation, obtaining a longer or complete virus sequence and clarifying the medical and epidemiological importance of the Dreznica virus.

Characterization of Carbapenemase-Producing Klebsiella pneumoniae in Clinical Center University of Sarajevo, Bosnia and Herzegovina.

Klebsiella pneumoniae is the second most prevalent gram-negative rod that causes nosocomial infections in hospitalized or otherwise immunocompromised patients. It can develop multiple drug resistance that results in limited treatment options and increased use of carbapenems. Various mechanisms are related to the development of carbapenem resistance in K. pneumoniae. The aim of this study was to perform phenotypic and molecular characterization of clinical isolates of carbapenemase-producing K. pneumoniae from two outbreaks recorded in 2017 and 2018 in Clinical Center University of Sarajevo, Bosnia and Herzegovina. Identification of K. pneumoniae isolates was carried out on the basis of morphological, cultural, and biochemical characteristics. Interpretation of antimicrobial resistance was performed according to EUCAST breakpoints. There were four different resistotypes of carbapenemase-producing K. pneumoniae in this study and all were confirmed positive for blaOXA-48 carbapenemase. Rep-PCR fingerprinting of these strains showed the presence of the two different genetic patterns with no similarity between them. The monitoring, surveillance, and molecular typing are essential to control the emergence of multidrug-resistant strains in nosocomial settings, and to reduce the frequency of outbreak occurrence.

Epidemiology of measles cases and phylogenetic analysis of the virus circulated in the Federation of Bosnia and Herzegovina during 2018: implications for elimination efforts.

Aim To present combined measles cases data and phylogenetic analysis of the virus circulated in 2018 in the Federation of Bosnia and Herzegovina (FB&H, the entity of Bosnia and Herzegovina), in order to analyse endemic transmission patterns of circulating strains and its implications for elimination efforts. Methods The data were derived from epidemiological case investigations and laboratory diagnoses based on serology, molecular detection and genotyping of the measles virus. Results During 2018 16 measles cases were reported in FB&H, of which five were classified as laboratory confirmed cases, one was an epidemiologically linked case and 10 were clinically compatible cases. Among them 12 (75.00%) cases were unvaccinated or had unknown vaccination status. The most affected population was up to 14 years of age (13/16; 81.25%). None of the cases was fully vaccinated. Viruses of other genetic lineages had been introduced in FB&H in the recent period. Two virus lineages of genotype B3 were identified. Phylogenetic analysis indicated the presence of a unique sequence of measles B3 virus in FB&H (Sarajevo). Conclusion Further strengthening of measles surveillance system and renewed efforts to increase vaccination levels are necessary to prevent disease and for elimination setting.

Genotyping test based on viral DNA, RNA or both as a management option for high-risk human papillomavirus positive women: a cross sectional study.

Aim This cross-sectional study of a group of women with abnormal cytology and high-risk human papillomavirus (hrHPV) infection compared genotyping HPV DNA and mRNA assays according to two age categories of women (S1: ≤30 and S2: >30 years). Methods The hrHPV DNA positive results of 105 cervical samples of women were pooled and those harbouring HPV-16, 18, 31, 33 and/or 45 DNA were tested for the type specific HPV oncogene E6/E7 overexpression (mRNA). Results Although HPV DNA testing showed a higher proportion of women infected by any of five hrHPVs in S1 group, total agreement of hrHPV DNA and mRNA positive results was higher in S2 group of women (75.8% v. 83.9%). The most prevalent type in both age groups was HPV-16. A 100% agreement of positivity of both tests was noted for HPV-18 and 33 in S1 group, and for HPV-18 in S2 group. Increasing concordance of HPV-16 and 31 DNA and mRNA positive results with the severity of cervical cytology was observed in S1 group of women. Absolute matching (100.0%) of positivity of both diagnostic tests was recorded in S2ASCUS group (for HPV-16, 18 and 33), in S1HSIL (for HPV-16, 18, 31 and 33), in S1LSIL category (for HPV-18 and 33) and in S2HSIL group (for HPV-18). Conclusion The results indicate the possibility of predicting the risk of persistent infection only by HPV DNA typing test, with no need for additional RNA testing in categories of infected women showing a high (absolute) agreement of positivity of both tests.

Antibiotic Resistance Profiles and Genetic Similarities Within a New Generation of Carbapenem-Resistant Acinetobacter calcoaceticus-A. baumannii Complex Resistotypes in Bosnia and Herzegovina.

Acinetobacter calcoaceticus-A. baumannii complex (ACB complex) is a nosocomial pathogen. Due to its high ability to develop antibiotic resistance, it has become a problematic challenge in the modern healthcare system. The molecular and genetic mechanisms of gaining multidrug resistance in ACB complex are well known. This study focuses on providing an overview of the antibiotic resistance profiles, genetic similarities and resistotypes, and general characteristics of carbapenem-resistant ACB complex (CRACB) in Bosnia and Herzegovina (BiH). In light of the data collected in this study, together with the already known information concerning antibiotic resistance of ACB complex, we intend to further elucidate the antibiotic therapy for CRACB strain resistotypes in BiH.

Comparison of the detection of HPV-16, 18, 31, 33, and 45 by type-specific DNA- and E6/E7 mRNA-based assays of HPV DNA positive women with abnormal Pap smears.

This study compares the type-specific human papillomavirus (HPV) DNA test with E6/E7 mRNA detection assay because of their importance in cervical cancer screening programs. A total of 105 women with positive high-risk Hybrid Capture 2 or Abbott RealTime High Risk HPV screening test and an abnormal cervical Pap smear were enrolled in the study. HPV typing was performed by multiplex real-time PCR (HPV High Risk Typing Real-TM test). HPV-16, 18, 31, 33, and 45 E6/E7 mRNAs were determined by type-specific real-time NASBA assay (NucliSENS EasyQ HPV v1.1). Infections caused by HPV-16, 18, 31, 33, and 45 types increased with severity of cervical cytology (p=0.008). Global positivity of five HPV E6/E7 mRNAs was lower than DNA positivity within women with atypical squamous cells of undetermined significance (p=0.016; p=0.008). High agreement of the tests was found in the groups of women with low-grade (p=1.000; p=0.063) and high-grade squamous intraepithelial lesion (p=0.250; p=0.125). Type-specific agreement of both diagnostic approaches was high regardless of cytology. Based on the found differences between HPV-16, 18, 31, 33, and 45 E6/E7 mRNA and DNA positivity, further study is needed to test the role of mRNA testing in the triage of women with atypical squamous cells of undetermined significance in Pap smear.

Surveillance of wildlife zoonotic diseases in the Balkans Region.

The countries of the Balkan Peninsula have become the region with frequent outbreaks of the emerging and re-emerging diseases during the last decade of the 20th and the first decade of the 21st century. The majority of outbreaks were wildlife zoonotic, and vector-borne diseases, such as brucellosis, leptospirosis, listeriosis, tularemia, Q-fever, Lyme disease, anthrax, rabies, viral hemorrhagic fevers, sandfly fever, tick-borne encephalitis and leishmainiasis. Epidemiological factors determined by ecology of causative agents are often the most useful diagnostic clues. The recognition of evolving problems of emerging and re-emerging diseases emphasizes the need for the development of better laboratory diagnostic methods for the surveillance and tracking of the diseases, and for continued research of factors contributing to the transmission of the organisms. The continuous occurrence of previously unidentified infections requires prospective national strategies for timely recognition of the syndromes, causative agent identification, establishment of criteria and methods for the diagnosis, optimization of the treatment regime, and determination of successful approaches to prevention and control. Wildlife diseases surveillance in the most of the Balkan countries has been coordinated by the WHO since 1992. Although new technology and communication have extremely improved in the last decade, there is a need for optimal communication lines among the Balkan countries, better exploitation of communication technologies like the Internet and other media in the field of emerging diseases.

Sandfly - Pappataci fever in Bosnia and Herzegovina: the new-old disease.

Sandfly fever viruses (SFV) are endemic in the Mediterranean, Middle East, northern African and western Asian countries. Toscana virus (TOSV), serotype of Sandfly fever Naples virus, is among of the three most prevalent viruses associated with meningitis during the warm seasons in northern Mediterranean countries. The historical data of the sandfly fever (Pappataci fever) indicates its origin in Bosnia and Herzegovina at the end of 19th century. There is a long period of time for which there are no data on research related to the SFV in Bosnia and Herzegovina. The purpose of the study was to investigate the presence of sandfly fever in Bosnia and Herzegovina in recent years. The 68 of serum samples were obtained from February 2006 until September 2008 from a group of patients with febrile illness of unknown etiology. The sera were tested on the presence of IgG and IgM antibodies against TOSV by specific serology test- recomLine Bunyavirus IgG/IgM immuno-line assay. The recent TOSV-infection was confirmed in the patients in each year during the study: 10,71% (3/28) in 2008; 9,38% (3/32) in 2007 and 12,50% (1/8) in 2006. The presence of specific antibodies to TOSV in the sera of the patients in recent years indicates re-emerging character of the disease in this region. It would be necessary to make biological, epidemiological and clinical research on the TOSV and related phleboviruses to elucidate the problem of SFV in Bosnia and Herzegovina.

Evaluation of hybrid capture 2 HPV DNA test and two variants of polymerase chain reaction (PCR-PGMY11/ PGMY09 and PCR-CPI/CPIIG) according to HPV types.

BACKGROUND: [corrected] The choice of diagnostic tool for the detection of oncogenic HPV types affects in a great manner on the uterine cervical carcinoma prevention in women with early stage of cervical dysplasia. METHODS: In this study 148 women with three subsequent cervical cytologic tests within two years showing mild dyskaryosis (Pap II) were enrolled. HPV infection was determined using three molecular approachs: Hybrid Capture 2 HPV DNA test and two variants of polymerase chain reaction (PCR-PGMY11/PGMY09 and PCR-CPI/CPIIG). HPV typing was performed by RFLP of PCR-PGMY11/ PGMY09 products. RESULTS: By HR-hc2 test, infection was detected in 32 of 38 HPV positive samples determined by minimally one approach. The results were negative for 6 samples with HPV infection caused by HPV6b, HPV44, HPV53 in 2/4 cases, HPV68 types and by multiple infection with four different HPV types: HPV66+HPV72+HPV73+HPV53, as well. Infections caused by low risk HPV types HPV6b and HPV44 were confirmed by using LR-hc2. CONCLUSIONS: hc2 HPV DNA test has satisfactory compatibility with both PCR variants according to HPV types. In the prevention of cervical carcinoma, high prevalence of new high risk HPV73 type and "probable high risk" HPV53 and HPV26 types in analyzed group of women should contribute to the fast production of new generation of the test enriched with RNA specific probes against mentioned oncogenic HPV types.

[HPV genotypes in repeated PAP II smears of group of women in Slovenia]. / HPV genotipovi kod grupe zena sa ponovljajucim PAP II rezultatom u Sloveniji.

Molecular epidemiology studies have shown that some types of HPV are causally linked with precancerous lesions of cervix and invasive cervical cancer. In our study, we enrolled HPV infected women with repeated Pap smears (Pap II) in Slovenia and verified HPV infection. HPV genotypes were determined using the method of enzyme restriction of PCR products amplified with group-specific oligonucleotide primers PGMY11/PGMY09 (PCR-RFLP) and restricted with seven different restriction endonucleases. In women aged < or = 30 years we detected 21 different types of HPV and a very high percentage (76.5%) of multiple infections. In women aged > 30 years we detected 11 different types of HPV and found the predominance of a single HPV type infection (85%). In women aged < or = 30 years the most frequent infections, and also equally distributed (10.50%), were those with high-risk types HPV16 and HPV73 and with a probably high-risk type HPV26. In women aged > 30 years most frequent infections, and also equally distributed (18.20%), were those with high-risk type HPV16 and probably high-risk type HPV53. The HPV genotypes were classified by the newest classification according to their oncogenic potential and we have referred the presence of newly high-risk and probably high-risk types of HPV at observed population of women in Slovenia. Using group-specific primers PGMY11/PGMY09 and seven restriction endonucleases in PCR-RFLP method for genotypization of HPV we identified single types of HPV included in large percentage of multiple infections of young women with repeated Pap II smears.

[Prevalence of human papillomavirus infection in Slovenian women with repeated Pap II smears]. / Prevalencija infekcije humanim papilomavirusima kod slovenackih zena sa ponavljajucim Pap II rezultatom.

In the detection of precancerous lesions the cervical Papanicolaou smear screening is used in Slovenia and worldwide. Management of patients with repeat abnormal smears (Pap II) represents a great and complex clinical and public health problem. Repeated cytologic examinations are routine procedure in many countries, also in Slovenia, although the sensitivity of Pap smear testing in the detection of cervical intraepithelial neoplasia (CIN) II and III is relatively low. In cases of abnormal squamous cells and mildly dyskaryotic cells the presence of infections with high-risk HPV genotypes is being increasingly used as a complementary method to Pap smear testing. In the study we enrolled 148 cervical samples of women who within two years had three subsequent Pap II smears (abnormal squamous cells or mildly dyskaryotic cells). The prevalence of HPV infections was determined using three molecular tests: Hybrid Capture 2 (hc2) test and two variants of polymerase chain reaction (PCR-PGMY11/PGMY09 and PCR-CPI/CPIIG). HPV infection was detected in 25.7% of women. In women aged < or = 30 years a statistically significant higher prevalence of HPV infections was found (37.8%) than in women aged > 30 years (20.4%). Our findings show that repeat Pap smear as the method of follow-up and detection of precancerous lesions of the observed population do not provide relevant results due to low prevalence of HPV infections in Slovenia, which indirectly indicates low sensitivity and specificity of Pap smear testing. In the detection of HPV infections, molecular methods are thus sensitive screening tests to be used complementary to cytologic tests in women with abnormal squamous cells and mildly dyskaryotic cells.