Long-Term Immunological Alertness and Response to COVID-19 Vaccination-Conditions for Prevention in Early Palliative Oncological Care Patients.

Aside from the general population, the COVID-19 pandemic has also affected a group of patients in palliative oncology care. In this study, long-term immune responses against SARS-CoV-2 after vaccination were monitored in a cohort of patients in palliative oncology care. This non-randomized, prospective, and open-label pilot study recruited patients from the Palliative Oncology Program and included 147 patients, of which 80 were females (54.4%) and 67 males (45.6%). The overall evaluation included current health status, SARS-CoV-2 anti-S IgG titer, and neutralizing antibodies using the SARS-CoV-2 virus neutralization test (VNT). Anti-S IgG antibody analysis revealed high (H) antibody levels in 35.7% (n = 10) and very high (VH) levels in 39.3% (n = 11) of patients after the second vaccination dose. Similarly, after the third dose, H was found in 29.6% (n = 32) and VH in 55.5% (n = 60) of patients. High and very high anti-S IgG antibody levels were consistent with high VNT titers (>2560) and H antibody levels in 17.1% (n = 12) or VH in 82.9% (n = 58) of patients. Patients with two or more doses showed H and VH antibody levels at a median of 451 and 342 days after vaccination, respectively. In this clinical trial, patients showed high and very high levels of anti-S IgG antibodies over a longer period of time. These patients did not show reduced immunological responses to the COVID-19 vaccine challenge. We can assume that prevention through vaccination can reduce the risk of complications or death from COVID-19 in patients in early palliative oncology care.

Multicentric evaluation of sensitivity of eight commercial anti-SARS-CoV-2 antibody assays and their correlation to virus neutralization titers in seropositive subjects.

Diagnosis of SARS-CoV-2 virus is mainly based on direct detection. Determination of specific antibodies has been used mostly for epidemiological reasons. However, select immunoassays showed good correlation to plaque reduction virus neutralization test (PRNT) in smaller patient cohorts, which suggests their potential as predictors of virus neutralization titer. A total of 3,699 samples from Covid-19 patients were included in the multicentric study performed in the Czech Republic. Anti-SARS-CoV-2 antibody levels were evaluated by 8 commercial antibody assays. Simultaneously, PRNT evaluations were performed with the SARS-CoV-2 B.1.258 variant. All immunoassays showed an overall high true positive diagnostic value ranging from 79.17 to 98.04%. Several commercial EIA methods showed highly positive correlation between the assay results and PRNT levels, e.g., Liaison CoV-2 TrimericS IgG DiaSorin (Spearman r = 0.8833; Architect SASRS-CoV-2 IgG Abbott (r = 0.7298); NovaLisa SARS-CoV-2 IgG NovaTec (r = 0.7103) and Anti-SARS-CoV-2 ELISA IgG Euroimmun (r = 0.7094). While this correlation was less positive for other assays, those, conversely, presented higher true positive values. For most immunoassays, the positive percent agreement of the results was ≥ 95% in sera exhibiting PRNT levels of 1:80 and higher. The assays tested have shown variable correlation to PRNT. Those possessing high positive predictive values serve well as qualitative tests, while others can be utilised as quantitative tests highly predictive of neutralization antibody levels.

The sensitivity and specificity of Abbott Panbio™ COVID 19 Ag Rapid test in the context of four SARS-CoV-2 variants.

Rapid antigen tests for the detection of SARS-CoV-2 are commonly used for the diagnosis of Covid-19. Previously published data showed a wide range of sensitivity and specificity of RATs, but these studies were performed on relatively small numbers of samples and using only limited numbers of virus variants. The aim of the study was to evaluate the main parameters of a commonly used RAT for 4 different virus variants in comparison with PCR. Material and methods: A set of 2874 samples obtained from Covid-19 patients were examined both by PCR and RAT. Two commercial PCR kits (Generi Biotech, Diana Biotechnologies) and one RAT - Abbott Panbio™ COVID 19 Ag Rapid - were compared for their sensitivity and specificity in samples positive for one of the four different SARS-CoV-2 variants - B.1.258 (n = 496), Alpha (n = 645), Delta/Delta+ (n = 687), and Omicron (n = 1046). Results: The sensitivity of Panbio™ COVID19 Ag Rapid test varied from 80.0 % in Omicron to 88.92 % in Alpha variants. The specificities of the RAT for all variants reached above 93 %. Statistically significant differences were found between the results from RAT assay in select virus variants. In addition, significantly higher sensitivity (p < 0.05) was detected in samples with higher viral loads than in those with lower. Conclusion: Despite the different sensitivity and specificity of Panbio™ COVID19 Ag Rapid test (Abbott ®) for different SARS-CoV-2 variants, this test sensitivity was proven to be always above the 80 % suggested by WHO, which makes it suitable for common use, regardless of the virus variability.

Long-Term Antibody Response and Vaccination Efficacy in Patients with COVID-19: A Single Center One-Year Prospective Study from the Czech Republic.

BACKGROUND: The diagnosis of SARS-CoV-2 is almost exclusively performed by PCR or antigen detection. The detection of specific antibodies has not yet been considered in official diagnostic guidelines as major laboratory evidence for a case definition. The aim the present study is to analyze antibody responses in outpatient and inpatient cohorts of COVID-19 patients in the Czech Republic over a 12-month period, and assess the potential of antibodies as a diagnostic tool. METHODS: A total of 644 patients was enrolled in the prospective study. IgA, IgM and IgG antibody levels, as well as virus neutralization titers, were analyzed over a 12-month period. RESULTS: Our study showed low antibody positivity levels at the admission. However, at 2 weeks after infection, 98.75% and 95.00% of hospitalized patients were IgA and IgG positive, respectively. Even in the outpatient cohort characterized by milder disease courses, the IgG antibody response was still sustained at 9 and 12 months. The data show a high correlation between the IgG levels and virus neutralization titers (VNTs). Samples from later time-points showed positive antibody responses after vaccination in both cohorts characterized by high IgG levels and VNT over 1:640. The samples from unvaccinated persons indicated a relatively high level of reinfection at 6.87%. CONCLUSIONS: Our results show that the detection of antibodies against the SARS-CoV-2 shows an increasing sensitivity from week 2 after infection and remains highly positive over the 12-month period. The levels of IgG antibodies correlate significantly with the VNTs. This suggests that the serological data may be a valuable tool in the diagnosis of SARS-CoV-2 infection.

Francisella tularensis caused cervical lymphadenopathy in little children after a tick bite: Two case reports and a short literature review.

Although Francisella (F.) tularensis is a well-described and understood zoonotic pathogen, its importance in Central Europe is relatively minor and, as such, tularaemia may be missed in the differential diagnosis. The annual incidence of tularaemia in the Czech Republic is relatively stable with up to 100 reported cases per year, except in the epidemic years 1998 and 1999 with 225 and 222 reported cases, respectively. It is, however, higher in comparison with the neighbouring countries. The common route of transmission in Central Europe is handling infected animals. Tularaemia is not commonly recognized as a tick-borne disease. Here we report two rare cases of a tick bite-associated ulceroglandular form of tularaemia in 2.5-year-old and 6.5-year-old children presenting with cervical lymphadenopathy. The unusual and interesting features of those cases are the young age and relatively uncommon route of transmission suggesting possible changes in the epidemiology of tularaemia in the Czech Republic. Therefore, the infection with F. tularensis should be considered in the differential diagnosis after a tick bite even in infants.

[Effect of the gut microbiota on the development of colorectal cancer and leakage of intestinal anastomoses].

Colorectal cancer is a very common malignancy with high mortality. Many factors influencing both the development and subsequent treatment, such as age, gender or genetic predisposition, are not modifiable. Others, such as stress, diet, physical activity or smoking, may be prevented by each individual. The gut microbiota is an important factor involved in both the development and treatment outcomes. With the advancing study of the gut microbiota, the relationship between its composition and various diseases is better understood. The proportions of members of the phyla Firmicutes (as beneficial microbiota) and Bacteroidetes (as mostly disease-associated microbiota) seem to be particularly important. Some studies suggest that certain bacteria may contribute to postoperative anastomotic leaks that prolong hospital stays, are a burden to patients, increase costs and may be fatal. Bacteria associated with the complication are, for example, enterococci, pseudomonads or bifidobacteria. Better understanding of the pathognomonic relationship between increased detection of certain bacteria and a complication may lead to individualized therapy aimed to reduce complications during surgical management of colorectal cancer.

Heterogeneity of post-COVID impairment: interim analysis of a prospective study from Czechia.

We stratified post-COVID patients into four newly established clinical groups based on the presence or absence of at least one subjective respiratory symptom and at least one objective sign of pulmonary involvement. Nearly half of outpatients and one third of hospitalized post-COVID patients had objective signs of pulmonary involvement without accompanying subjective respiratory symptoms three months after diagnosis.

Effective Treatment for Uncomplicated Urinary Tract Infections with Oral Fosfomycin, Single Center Four Year Retrospective Study.

Fosfomycin represents a relatively old antibiotic, but it is experiencing a comeback in recent years. According to some studies, the increasing therapeutic use of this drug led to a rapid increase in the levels of resistance in bacteria causing urinary tract infection. In the presented study, levels of resistance to fosfomycin in more than 3500 bacterial isolates before and after fosfomycin introduction into therapeutic use in the Czech Republic and the clinical efficacy of treatment in 300 patients using this drug were assessed. The results show that the resistance levels to fosfomycin in Escherichia coli isolates before and after the drug registration were not significantly different (3.4% and 4.4%, respectively). In some other Gram-negative rods, such as otherwise susceptible Enterobacter, resistance to fosfomycin increased significantly from 45.6% to 76.6%. Fosfomycin treatment of urinary tract infections showed an excellent seven-day clinical efficacy (79.7%). However, when used to treat recurrent or complicated urinary tract infections, fosfomycin treatment was associated with high levels of infection relapse, leading to relapse in a total of 20.4% of patients during the first two months. This indicates that fosfomycin exhibits good efficacy only for the treatment of uncomplicated urinary tract infections.

Isolation of Bordetella trematum from the respiratory tract of a patient with lung cancer: a case report.

We report the case of isolation of Bordetella trematum from the respiratory tract of a patient with lung carcinoma. This gram-negative, opportunistic rod was firstly described in 1996. To date, only several strains of Bordetella trematum have been isolated and reported, mostly from skin and soft tissue infections. The patient was admitted to the ICU of the Pulmonary Department in incipient septic shock with respiratory failure. Intravenous fluid resuscitation and non-invasive ventilation were administered immediately. A broad spectrum antibiotic piperacillin/tazobactam was administered empirically after sampling of material for microbiological examination. The bronchoscopy showed a large cavern of decayed tumour invading into mediastinum. Both sample cultures showed significant quantities of gram-negative non-fermenting bacteria. The isolate was identified using MALDI-TOF MS as Bordetella trematum and the identification was confirmed using 16S ribosomal RNA sequencing. In the last few years, routine bacterial identification using MALDI-TOF MS has enabled correct discrimination of this species. Nevertheless, isolation of Bordetella trematum in clinical samples is still very uncommon, and it is appropriate to confirm the species identification via 16S ribosomal RNA sequencing. To our knowledge, this is the first case of B. trematum isolated from the human respiratory tract since its first description. The clinical significance of Bordetella trematum in the rapid deterioration of the patient's status remains unclear.

Herpes simplex virus resistant to acyclovir: A single-centre experience from the Czech Republic.

INTRODUCTION AND AIM: Infections caused by herpes simplex viruses (HSV) are frequent in the human population. Because of the widespread use of long-term treatment or prophylaxis by anti-herpetic antivirals in various specific medical contexts (immunosuppression, recurrent infections), the level of antiviral resistance is increasing. According to previous studies, there is a low resistance level in immunocompetent populations but a relatively high level in populations with immunodeficiency. However, there has been no study from the Czech Republic. This study presents results of a single-centre retrospective study from the Czech Republic. MATERIALS AND METHODS: Deep frozen DNA from patients with suspected clinical antiviral failure over a long time period (2009-2016) - a total of 15 isolates of HSV1 and seven of HSV2 - were examined for the presence of mutations associated with antiviral resistance. Sequence analysis was performed using an ABI PRISM 3500xL Genetic Analyzer (Applied Biosystems®). RESULTS: There were no mutations associated with resistance to antivirals inside the UL23 gene in HSV1 isolates. However, resistant mutation D672N (nucleotide change G2014A) was found inside the UL30 gene in seven of the isolates. One mutation associated with resistance to acyclovir (M183stop) was found inside the UL23 gene in one HSV2 isolate. Resistant mutation E678G (nucleotide change A2033G) was identified inside the UL30 gene in six of the HSV2 isolates. CONCLUSIONS: This study confirmed the presence of resistance mutations within the Czech population, but it will be necessary to examine a higher number of isolates for further conclusions.

Human polyomavirus 9 in immunocompromised patients in the University Hospital in Hradec Kralove, Czech Republic.

Human polyomaviruses such as JC polyomavirus and BK polyomavirus have long been well known pathogens of immunocompromised patients. Several new members of this viral family have been described during the last decade. Human polyomavirus 9 seems to be a novel pathogen of transplanted patients according to some studies. The aim of our study was to determine the presence of human polyomavirus 9 in patients after kidney or stem cell transplantation (SCT) at the University Hospital in Hradec Kralove, Czech Republic. Overall 100 patients, 65 after kidney transplantation and 35 after SCT, were included into the study. At least three follow-up samples from each patient were examined for human polyomavirus 9 DNA presentation with the two previously described in-house PCR protocols. Despite the frequent reactivation of human CMV (14.3% in kidney transplantation and 63.3% after SCT) or BK polyomavirus in our patient group, there was no positivity for human polyomavirus 9 either in blood samples or urine samples. One of the possible reasons for this discrepancy versus previous published studies could be a relatively low proportion of patients treated by induction therapy before kidney transplantation in our study cohort.

Mumps in the Eastern Bohemia Region of the Czech Republic - a Serological Survey 2008-2012.

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.

The susceptibility to fosfomycin of Gram-negative bacteria isolates from urinary tract infection in the Czech Republic: data from a unicentric study.

BACKGROUND: Against a background of rapid increase of ß-lactamase-producing or multi-resistant pathogenic bacteria and the resulting lack of effective antibiotic treatment, some older antibiotics have been tested for new therapeutic uses. One of these is fosfomycin, to which according to studies these resistant bacteria are very sensitive. Our study was designed because there is no data on the fosfomycin susceptibility rate in the Czech Republic. METHOD: In this study from January 2013 to June 2014 3295 unique isolates of Gram-negative bacteria which had caused urinary tract infections were examined. The antibiotic susceptibility was measured by disk diffusion test. Both EUCAST and CLSI guidelines criteria (for fosfomycin only) were used for the antibiotic susceptibility evaluation. RESULTS: The most frequently tested bacterial isolates were Escherichia coli (51.3%, n = 1703), Klebsiella pneumoniae (19.4%, n = 643) and Proteus spp. (11.8%, n = 392). Among all isolates 29.0% (n = 963) were resistant to fluoroquinolones, 11.3% (n = 374) produced extended spectrum ß-lactamase and 4.2% (n = 141) produced AmpC ß-lactamase. The overall in vitro susceptibility was significantly higher for fosfomycin compared to the other tested per-oral antibiotics (nitrofurantoin, ampicillin, co-trimoxazole, ciprofloxacin and cefuroxime) against all tested Gram-negative rod isolates (excluding Morganella morgani and Acinetobacter spp. isolates). Fosfomycin also remained highly active against those isolates with extended spectrum ß-lactamase (ESBL) production (95.8% in Escherichia coli isolates and 85.3% in Klebsiella pneumoniae isolates), unlike other tested per-oral antibiotics, which showed significant (p < 0.0001) susceptibility decrease. CONCLUSION: We have confirmed in the Czech Republic the very high susceptibility to fosfomycin trometamol of urinary tract infection pathogens, particularly Gram-negative rods including those producing ß-lactamase.

Prevalence of overweight and obesity in professional soldiers of the Czech Army over an 11-year period.

BACKGROUND/AIM: Obesity is currently considered to be the most frequent metabolic disease worldwide, not only in developed but also in developing countries. The aim of this work was to describe the development of health status in soldiers of the Armed Forces of the Czech Republic (ACR) and to emphasizethe markers of non-communicable diseases. Our study describes the anthropometric and biochemical parameters of a large group of Czech Army professional soldiers. Data were obtained over a period of 11 years. METHODS: During the monitored period, from 1999 to 2009, military physicians carried out on the average 6,360 examinations on professional soldiers per year and monitored their health and nutritional status with the aim of preventing the risk factors of non-communicable diseases. These examinations are compulsory for all professional soldiers at the age of 25, 30, 33, and 36 years. From the age of 39, these examinations are carried out every year till the end of their career. Besides taking personal histories and carrying out standard physical examinations, blood was taken for biochemical examination. The following anthropometric parameters were monitored: body constitution using body mass index (BMI) and waist circumference. Our study describes only part of the data concerning anthropometric and biochemical parameters of professional soldiers which were obtained over a period of 11 years. RESULTS: Average BMI values in men were in the overweight range (26.5-27 kg/m2). Average values of waist circumference, however, ranged from 91.9 cm to 93.4 cm. Between the first and the last year of monitoring a statistically significant decrease in these values ranging from 93.4 ± 9.8 cm to 92.7 ± 9.5 cm (p < 0.001) was observed. All monitored anthropometric parameters in female professional soldiers were within normal limits. During the monitored period the proportion of overweight men gradually increased from 52% to 57.1% (p < 0.001). There were no statistically significant changes in the prevalence of obesity in men (12-15%). Average glycaemia levels were within normal range in both men and women. A statistically significant decrease in these levels, however, was observed in men (from 5.1 ± 0.9 mmol/L to 4.8 ± 0.7 mmol/L (p < 0.001) and in women (from 4.9 ± 0.6 mmol/L to 4.6 ± 0.6 mmol/L (p < 0.001). Concerning the lipid profile in men, a significant decrease in average values of total cholesterol, triglycerides and LDL cholesterol was observed; from 5.5 ± 1.1 mmol/L to 5.1 ± 1.0 mmol/L (p < 0.001), from 2.0 ± 1.6 mmol/L to 1.6 ± 1.2 mmol/L (p < 0.001) and from 3.4 ± 1.1 mmol/L to 3.2 ± 0.9 mmol/L (p < 0.001), respectively. CONCLUSION: During the monitored period only one-third of military professionals had normal body weight. More favorable situation was in female professional soldiers, two-thirds of them had normal body weight during the monitored period. Additionally, the increase in the number of individuals with BMI values in the overweight range was observed. Although the number of overweight soldiers was overestimated as a result of the inclusion of individuals with increased body weight due to well-developed musculature, the number of overweight and obese soldiers is still high.

[Human polyomaviruses]. / Lidské polyomaviry.

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.

[Viral gastroenteritis]. / Virové gastroenteritidy.

The causative agents of viral gastroenteritis belong to generally little known pathogens. But according to some studies, these viruses make up a considerable proportion of agents causing diarrhea in children and adults. In this review, the authors focused on both relatively known pathogens such as adenoviruses, rotaviruses or caliciviruses and not so well known or newly recognized viral gastroenteritis pathogens like toroviruses, astroviruses, bocaviruses or parechoviruses. The first part of this review is concerned with basic information about each etiological agent. The second one deals with their detection and identification. The last part is about therapy and prophylaxis of viral pathogens.

[Options for the detection and identifications of agents causing viral hemorrhagic fevers]. / Moznosti detekce a identifikace puvodcu virových hemoragických.

Viral hemorrhagic fevers (VHFs) belong to the most deadly diseases. Therefore rapid detection and identification of causal agent is very important for epidemiological investigation and countermeasures. The review present all published information about VHF detection and identification, with focus on polymerase chain reaction (PCR) methods. In the first part, basic information about microbiological diagnostic technique is provided. The second part is concerned with detection and identification methods for individual agents causingt VHFs.

[Experience with the PCR method in chlamydia detection.].

PURPOSE OF THE STUDY: The study was intended to summarize and evaluate the results in patients with a suspected infection by the genus Chlamydia, investigated with an in-house method of nested PCR (polymerase chain reaction). The study worked with data from patients living in eastern Bohemia, who were examined in the years 2001-2003 at the Dept. of Molecular Biology, a research laboratory shared by the Institute of Clinical Microbiology and the Institute of Clinical Biochemistry and Diagnostics. MATERIAL AND METHODS: 291 explorations were done in 2001, in 2002 already 562 and in 2003 their figure reached 760. The total number of samples received for investigation during that period was 1 613. 1 587 were actually investigated, 26 were unsuitable and could not be used. More than 70 % of all investigations were done with three types of material: urine (41.8 % of all the investigated samples), BAL (15.3 % of all the investigated samples) and whole blood (14.9 % of all the investigated samples). The investigations were carried out with the in-house nested PCR method, which uses primers from the MOMP(ompA) area of the genus Chlamydia spp. RESULTS: Total positivity was 5.67 %, in 1.26 % of the samples the resulted was considered uncertain and 93.07 % of the investigated samples were negative. In men PCR positivity was 6.11 %, in women 5.35 %. The major proportion of positive samples was from the age groups 70-79 years (11.67 %), 10-19 years (6.51 %) and 40-49 years (6.45 %). Overall positivity in smears from the urogenital system was 6.48 %, from urine 3.92 %, from BAL 10.70 % and from whole blood 5.51 %. KEYWORDS: nested PCR, Chlamydia spp., detection.