The optimal dosage of anti-thymocyte globulin (ATG) may influence the outcome of patients after allogenic haematopoietic stem cell transplantation (HSCT). The aim of our study was to analyse human cytomegalovirus (CMV) infection data, incidence of graft-versus-host disease and other clinical endpoints comparing two patients cohorts that were administered two different Thymoglobuline Genzyme doses as part of the HSCT conditioning regimen.Total of 65 adult patients received ATG (7.5 mg/kg or 6 mg/kg) as a part of the fludarabine/busulfan/ATG conditioning regimen. CMV DNAemia was monitored after HSCT using quantitative real-time PCR and preemptive treatment was started for viral loads above 1000 cp/ml.The mild ATG dose reduction extended the time to the first CMV detection after transplantation (28 days for 7.5 mg/kg dose vs. 40 days for 6 mg/kg dose, p = 0.04). But it did not reduce the incidence or influence first anti-CMV treatment onset, the initial viral load, peak viral load in whole blood or the antiviral therapy parameters (all p 0.18). No impact of ATG dose reduction on incidence of graft-versus-host-disease, relapse of underlying disease or mortality within first year after transplantation (all p 0.32) were observed.The reduced ATG dosages can allow lower toxicity of conditioning regimen while keeping the performance.

Introduction: The optimal dosage of anti-thymocyte globulin (ATG) may influence the outcome of patients after allogenic haematopoietic stem cell transplantation (HSCT). The aim of our study was to analyse human cytomegalovirus (CMV) infection data, incidence of graft-versus-host disease and other clinical endpoints comparing two patients' cohorts that were administered two different Thymoglobuline Genzyme doses as part of the HSCT conditioning regimen. Materials and Methods: Total of 65 adult patients received ATG (7.5 mg/kg or 6 mg/kg) as a part of the fludarabine/busultan/ATG conditioning regimen. CMV DNAemia was monitored after HSCT using quantitative real-time PCR and preemptive treatment was started for viral loads above 1000 cp/ml. Results: The mild ATG dose reduction extended the time to the first CMV detection after transplantation (28 days for 7.5 mg/kg dose vs. 40 days for 6 mg/kg dose, p = 0.04). But it did not reduce the incidence or influence first anti-CMV treatment onset, the initial viral load, peak viral load in whole blood or the antiviral therapy parameters (all p >= 0.18). No impact of ATG dose reduction on incidence of graft-versus-host-disease, relapse of underlying disease or mortality within first year after transplantation (all p >= 0.32) were observed. Conclusions: The reduced ATG dosages can allow lower toxicity of conditioning regimen while keeping the performance.

The present study aims to evaluate the diagnostic yield of bronchoalveolar lavage (BAL) fluid in patients with hematological malignancies and describe the most common pathogens detected in BAL fluid (BALF.) An analysis of 480 BALF samples was performed in patients with hematological malignancies over a period of 7 years. The results of culture methods, PCR, and immunoenzymatic sandwich microplate assays for Aspergillus galactomannan (GM) in BALF were analyzed. Further, the diagnostic thresholds for Aspergillus GM and Pneumocystis jiroveci were also calculated. Microbiological findings were present in 87% of BALF samples. Possible infectious pathogens were detected in 55% of cases; 32% were classified as colonizing. No significant difference in diagnostic yield or pathogen spectrum was found between non-neutropenic and neutropenic patients. There was one significant difference in BALF findings among intensive care units (ICU) versus non-ICU patients for Aspergillus spp. (22% versus 9%, p = 0.03). The most common pathogens were Aspergillus spp. (n = 86, 33% of BAL with causative pathogens) and Streptococcus pneumoniae (n = 46, 18%); polymicrobial etiology was documented in 20% of cases. A quantitative PCR value of > 1860 cp/mL for Pneumocystis jirovecii was set as a diagnostic threshold for pneumocystis pneumonia. The absorbance index of GM in BALF of 0.5 was set as a diagnostic threshold for aspergillosis. The examination of BAL fluid revealed the presence of pathogen in more than 50% of cases and is, therefore, highly useful in this regard when concerning pulmonary infiltrates.

Chronic hepatitis C is curable disease. Low detection rate could be one of the reasons of poor treatment uptake. It is important to identify HCV prevalence and anti-hepatitis C virus (HCV) positive patients in population by effective screening strategy such as risk-based or birth cohort screening programs. There are no national population-based estimates of the HCV prevalence in the Czech Republic (CZ). The most recent seroprevalence survey determined a prevalence of positive anti-HCV antibodies of 0.2% (in 2001). The aim of the study was to determine the seroprevalence of HCV, HCV viraemia and HCV genotype in the CZ adult population. We also estimated the number of persons living with chronic hepatitis C in CZ. The examined group included 3000 adults, 18-90 years of age enrolled in 2015. All serum samples were examined to determined anti-HCV antibodies positivity, HCV-RNA positivity and genotypes. Of the 3000 samples, 50 were found to be anti-HCV-positive, for a seroprevalence of 1.67% (2.39% in males, 0.98% in females). The overall prevalence of positive HCV RNA was 0.93%: 1.5% in males, 0.39% in females. HCV genotype (GT) 1a was determined in 25%, GT 1b in 25% and GT 3a in 46%. Since 2001, the HCV seroprevalence has increased 8-fold. The highest HCV seroprevalence occurred in males aged 30-44 years. We can estimate that there are more than 140,000 people with HCV antibodies and more than 80,000 people with chronic hepatitis C living in the CZ. The introduction of birth cohort HCV screening could be beneficial for the country.

Herpetické viry patří mezi významné patogeny v imunosuprimované populaci, což vede ke stoupající spotřebě antiherpetik a také ke stoupajícímu problému se vznikem rezistence k těmto lékům.Článek stručně pojednává o nejvýznamnějších zástupcích z antiherpetik, o základních mechanismech vzniku rezistence a u vybraných herpetických virů také o současných znalostech

BACKGROUND:The incidence of mumps has decreased in many countries since the introduction of vaccination programmes, however, in the past decade a rapid increase in the disease occurrence has been reported worldwide. The reason for this situation is still not clear. We present the results of a serological survey carried out in the Eastern Bohemia Region of the Czech Republic during the years 2008-2012.METHODS:In total, 2,536 samples of 2,034 patients were examined during the study period. The study cohort was divided into two groups, one consisted of individuals born before the introduction of mandatory vaccination and the other one comprised individuals born after mandatory vaccination started. For the serology analyses the ELISA kits RIDASCREEN Mumpsvirus IgM and IgG (R-Biopharm®, Germany) were used.RESULTS:Out of 2,536 samples (including paired sera), 23.9% (n=606) were positive and 12% (n=304) had equivocal results. Most of the positive samples were obtained from patients aged 17-20 years. Significantly more (p<0.05) positive patients were born after the start of the national vaccination programme (patient group 2) (22.8%) compared to those born before its start (patient group 1) (13.7%). Interestingly, the analysis of data showed that 75.3% of patients falling into group 1 had anti-mumps IgG antibodies, which means that they had contracted mumps, whilst 23.5% of patients of group 2 had undetectable IgG antibodies, even though they should have been vaccinated.CONCLUSION:The data from our study, with a low number of positive samples in the first years of the study and an increase in the last two years, could suggest the occurrence of outbreaks every 4-6 years.

OBJECTIVES:The aim was to introduce a diagnostic method for detecting variants of hepatitis C virus (HCV) with protease NS3 resistance primarily to simeprevir (Q80K mutation in HCV genotype 1a) and its subsequent use in routine practice.MATERIAL AND METHODS:The detection of HCV resistance-associated variants in the NS3 protease gene by sequence analysis was introduced in the molecular biology laboratory of University Hospital Hradec Kralove in 2015. The primers were designed by sequence analysis software Custom Primers - OligoPerfect™ Designer. The method was optimized for HCV genotype 1a. The search for variants was performed using two programs.RESULTS:A total of 16 patients with genotype 1a chronic hepatitis C have been examined since 2015. In five of them, the Q80K variant was detected.CONCLUSION:The development of resistance to antiviral therapy for chronic hepatitis C gained importance after the introduction of direct-acting antivirals. Given the relatively high prevalence of the Q80K mutation in HCV genotype 1a, it is crucial to confirm its presence or absence before the therapy is initiated. The reported method enables clear and early detection of the Q80K mutation.

OBJECTIVE:To determine the incidence of infection with ganciclovir-resistant cytomegalovirus (CMV) in adult allogeneic hematopoietic stem cell transplant (HSCT) recipients. Clinical resistance or treatment failure was defined as persistent DNAemia or increasing viral load in peripheral blood after 2 weeks of virostatic treatment. The association between the treatment failure and viral resistance was analysed. The presence of ganciclovir-resistant CMV strains was confirmed by genotypic testing able to detect mutations conferring resistance.METHODS:In 2012 and 2014, 40 patients who underwent allogeneic HSCT for hematologic malignancies and were treated for human CMV reactivation/disease were followed up prospectively. In patients with treatment failure, CMV DNA was isolated and analysed by nucleotide sequence analysis of the UL 97 and UL 54 genes conferring resistance to the virostatic agent.RESULTS:The treatment failure occurred in seven patients, but ganciclovir resistance conferring mutations were only detected in two of them (mutations L595F and M460I in the UL 97 gene). Another mutation in the UL 97 gene (N510S) was found in a patient with recurrent CMV replication who needed to be retreated but did not meet the criteria for treatment failure.CONCLUSION:The low incidence of genetically confirmed ganciclovir-resistant CMV isolates in HSCT recipients with relatively common clinical treatment failure suggests that the mechanism underlying slower viral clearance is often other than mutations conferring ganciclovir resistance to the virus.

No: 1501 Presentation at ESCV 2015: Poster 2 The study of resistance of human cytomegalovirus to ganciclovir in patients after alogeneic transplantation of haematopoietic stem cells Stepanova Vlasta1, L. Pliskova2, E. Vejrazkova3, R. Kutova2, P. Hubacek4, M. Fajfr1 1 Inst. of Clin. Microbiology, University Hospital and Faculty of Medicine, Charles University, Hradec, Czech Republic 2 Inst. of Clin. Biochemistry and Diagnostics, University Hospital and Faculty of Medicine, Charles University, Hradec, Czech Republic 3 Clinic of Internal Diseases, Department of Clin. Haematology, University Hospital and Faculty of Medicine, Charles University, Hradec, Czech Republic 4 Clinic of Paediatric Haematology and Oncology, Inst. of Medical Microbiology of University Hospital,

Polyomaviruses belong to a group of viruses that has recently attracted the attention of many research groups. During 35 years, JC and BK viruses, known pathogens in immunocompromised patients, seemed to be the only human polyomaviruses. But in 2007, two other polyomaviruses, WU and KI, were isolated whose pathogenicity is still a matter of discussion. A year later, another human polyomavirus, associated with Merkel cell carcinoma, was identified, and seven more were described by the end of 2014. Some of them were found to be related to various diseases, others seem to be a part of the normal skin and mucosal microbiome. The article summarizes basic information about all so far described human polyomaviruses.

PURPOSE:Acute gastroenteritis is one of the most common diseases in humans worldwide and represents a significant cause of mortality and morbidity. The majority of cases are of viral aetiology, evidence for which has been increasing in the past decade. Several studies on the prevalence in European countries of viral aetiology of gastroenteritis have been published in the last decade, but none from the Czech Republic.MATERIAL AND METHODS:In total 107 faeces samples obtained from patients hospitalised in the University Hospital in Hradec Králové were examined by immunochromatographic tests using ROTA-ADENO Card Rapid-Viditest (VIDIA, Czech Republic) and RidaQuick Norovirus (R-Biopharm, Germany), and by an in-house Real-time PCR panel.RESULTS:Overall findings of viruses detected by PCR in the tested faeces samples were: rotaviruses in 29.9%, noroviruses in 14.0% and adenoviruses in 5.0%. Immunochromatographic antigen detection performed at lower sensitivity compared with PCR: rotaviruses in 28.0%, noroviruses in 4.7% and adenoviruses in 2.0%. Our findings demonstrate even lower sensitivity of the used immunochromatographic tests compared with manufacturers data.CONCLUSION:Our study has revealed limitations in immunochromatographic tests, especially in their sensitivity and the necessity for using another confirmatory method. We have set up real-time PCR in routine diagnosis of viral gastroenteritis in our hospital.

Purpose: The evaluation of four post-pandemic influenza seasons 2009-2013 in the Faculty hospital Hradec Kralove and comparison of used rapid antigen tests (RATs) with real time RT-PCP.Material and methods: Between November 2009 and June 201.3 were examined 3845 samples from patients with respiratory tract infections by RATs (Influenza A/B 2 Panel Test (GECKO '" Pharma, Germany), Rapid VIDITEST Influenza A+B Card (VIDIAJ. Czech Republic) and BinaxNOW Influenza A&B Test (ALERP, USA) or real time RT-PCP (RIR InfA/H1N1 Detection Set (Roche'"), RealStar" Influenza S and T PT-PCR Kit 3.0 (Altona, Germany).Results: A totally 1059 samples were examined simultaneously by RAT and real time RT-PCR. The overall sensitivity and specificity of RATs compared with real time RT-PCR were 32,2% and 98,1% for influenza A and 17.6% and 99.4% for influenza B. Higher sensitivity of RATs were in children (66.6%) compared with adults (14.3 - 40.0 %). In the first three post-pandemic seasons were continuously decrease of positive samples from 23.5% in season 2009-2010 to 3.3% in season 2011-2012. but in season 2012-2013 were rapidly increase of positive results. to 31.5%, with high share of influenza A/ H1N1/ 2009 (79,6%).Conclusion: Our results shown insufficient sensitivity of all used PATs and necessity of having other confirmatory test, like RT-PCR. It was also shown unexplained increase of case and influenza severity in season 2010-0013.

The epidemiology of selected sexually transmitted diseases in the Czech Republic has been carefully evaluated for many years. Data from 1981-2011 for eastern Bohemia shows a sharp decrease in the incidence of gonorrhea in 1993-1994 and a very low incidence thereafter with a slightly higher prevalence in males. However, syphilis and genitourinary infections with Chlamydia trachomatis show entirely opposite trends. Also, for the similar number of diagnostic tests performed, Chlamydia had a 10 fold higher rate of positive results. This underscores the changing epidemiology of sexually transmitted infections (STI) and necessity for adapting the reporting algorithms accordingly.