ABSTRACT
BACKGROUND: Opioids and epidural analgesia are a mainstay of perioperative analgesia but their influence on cancer recurrence remains unclear. Based on retrospective data, we found that cancer recurrence following colorectal cancer surgery correlates with the number of circulating tumor cells (CTCs) in the early postoperative period. Also, morphine- but not piritramide-based postoperative analgesia increases the presence of CTCs and shortens cancer-specific survival. The influence of epidural analgesia on CTCs has not been studied yet. METHODS: We intend to enroll 120 patients in four centers in this prospective randomized controlled trial. The study protocol has been approved by Ethics Committees in all participating centers. Patients undergoing radical open colorectal cancer surgery are randomized into epidural, morphine, and piritramide groups for perioperative analgesia. The primary outcome is the difference in the number of CTCs in the peripheral blood before surgery, on the second postoperative day, and 2-4 weeks after surgery. The number of CTCs is measured using molecular biology methods. Perioperative care is standardized, and relevant data is recorded. A secondary outcome, if feasible, would be the expression and activity of various receptor subtypes in cancer tissue. We intend to perform a 5-year follow-up with regard to metastasis development. DISCUSSION: The mode of perioperative analgesia favorably affecting cancer recurrence would decrease morbidity/mortality. To identify such techniques, trials with long-term follow-up periods seem suboptimal. Given complex oncological therapeutic strategies, such trials likely disable the separation of perioperative analgesia effects from other factors. We believe that early postoperative CTCs presence/dynamics may serve as a sensitive marker of various perioperative interventions´ influences on cancer recurrence. Importantly, it is unbiased to the influence of long-term factors and minimally invasive. Analysis of opioid/cannabinoid receptor subtypes in cancer tissue would improve understanding of underlying mechanisms and promote personalization of treatment. We are not aware of any similar ongoing studies. TRIAL REGISTRATION NUMBER: NCT03700411, registration date: October 3, 2018. STUDY STATUS: recruiting.
Subject(s)
Analgesia, Epidural , Colorectal Neoplasms , Neoplastic Cells, Circulating , Humans , Analgesics, Opioid/therapeutic use , Prospective Studies , Retrospective Studies , Neoplasm Recurrence, Local , Morphine , Colorectal Neoplasms/surgery , Randomized Controlled Trials as Topic , Multicenter Studies as TopicABSTRACT
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.
Subject(s)
COVID-19 , Lung/physiopathology , COVID-19/complications , COVID-19/epidemiology , COVID-19/pathology , Czech Republic/epidemiology , Hospitalization , Humans , Prospective Studies , SARS-CoV-2 , Post-Acute COVID-19 SyndromeABSTRACT
BACKGROUND: Invasive infections caused by Capnocytophaga canimorsus are rare. Immunocompromised patients, who report being bitten by or having a close contact with an animal, represent a high-risk group for this infection. There are only few dozens of infections by this bacteria manifesting as purulent meningitis reported worldwide. The reported case is a first reported case of purulent meningitis caused by by Capnocytophaga canimorsus in Czech Republic with only a limited risk factor history. CASE PRESENTATION: The patient, a 74 years old man, was referred to the infectious diseases department of a teaching hospital with clear signs of developing purulent meningitis. His anamnestic data did not show any unusual findings. He was treated for compensated diabetes mellitus type II. The blood cultures were negative and the etiological agent did not grow from the cerebrospinal fluid (CSF) on common media. Eventually, it was identified by detecting pan-bacterial DNA and DNA sequencing. Subsequently, the pathogen was confirmed by anaerobic cultivation from CSF. Only after then the patient recalled being bitten by his German shepherd puppy during play. The patient was successfully treated intravenously by ceftriaxone. CONCLUSIONS: Purulent meningitis caused by Capnocytophaga spp. is a rare disease, but it needs to be considered in patients at risk with pre-existing conditions, who report close contact with or being bitten by an animal. It is important to test for this microbe in cases with negative microbiological results for the more common agents.
Subject(s)
Capnocytophaga/pathogenicity , Gram-Negative Bacterial Infections/microbiology , Meningitis, Bacterial/microbiology , Aged , Animals , Bites and Stings/complications , Blood Culture , Capnocytophaga/genetics , Ceftriaxone/therapeutic use , Czech Republic , Dogs , Gram-Negative Bacterial Infections/drug therapy , Gram-Negative Bacterial Infections/etiology , Humans , Immunocompromised Host , Male , Meningitis, Bacterial/drug therapy , Meningitis, Bacterial/etiologyABSTRACT
Definitive diagnosis and therapy proved challenging in the case of a 60-year-old male with malaria and rickettsiosis. Returning travellers who are unwell can present practical difficulties in diagnosis and treatment and the focus here is on conditions relevant to the Republic of South Africa. Malaria, rickettsiosis and Q fever are discussed.
Subject(s)
Malaria/diagnosis , Rickettsia Infections/diagnosis , Travel , Humans , Malaria/complications , Malaria/epidemiology , Male , Middle Aged , Rickettsia Infections/complications , Rickettsia Infections/epidemiology , South Africa/epidemiologyABSTRACT
Studies focused on arbovirus diseases transmitted by invasive species of mosquitoes have become increasingly significant in recent years, due to the fact that these vectors have successfully migrated to Europe and become established in the region. Mosquitoes, represented by more than 3 200 species, occur naturally worldwide, except in Antarctica. They feed on the blood of warm-blooded animals and by this route, they are capable of transmitting dangerous diseases. Some species can travel a distance of 10 km per night and can fly continuously for up to 4 hours at a speed of 1-2 km/h. Most species are active at night, in the evening or morning. It usually takes a mosquito female about 50 seconds to penetrate the skin of mammals and the subsequent blood meal usually takes about 2.5 minutes. Mosquitoes live for several weeks or months, depending on the environmental conditions. The VectorNet project is a European network of information exchange and sharing of data relating to the geographical distribution of arthropod vectors and transmission of infectious agents between human populations and animals. It aims at the development of strategic plans and vaccination policies which are the main tasks of this time, as well as the development and application of new disinfectants to control vector populations.
Subject(s)
Communicable Diseases/transmission , Culicidae , Insect Vectors , Introduced Species , Animals , Biological Factors , Humans , VaccinationABSTRACT
AIM: To compare the effectiveness of treatment for ocular toxoplasmosis with pyrimethamine + clindamycin (or sulfadiazine) + a corticoid (Group 1), or azithromycin or a combination of azithromycin with a corticoid or a corticoid alone (Group 2). To determine the relapse rate depending on the treatment approach. MATERIAL AND METHODS: A total of 25 patients treated for ocular toxoplasmosis over the last five years (2008-2013) were analyzed. Group 1 comprised 16 patients (3 were excluded) and Group 2 consisted of 6 patients. RESULTS: Visual improvement was more rapid in Group 1 (day 10.7) than in Group 2 (significant improvement on day 29.6). There were 5 cases of relapse in Group 1; in 13 cases, no relapse was noted; all patients in Group 2 relapsed (a total of 13 relapses). Twenty-three patients were positive for specific IgG antibodies. CONCLUSION: According to our experiences, pyrimethamine + clindamycin (or sulfadiazine) + a corticoid should be the treatment of choice in patients with ocular toxoplasmosis.
Subject(s)
Adrenal Cortex Hormones/therapeutic use , Anti-Bacterial Agents/therapeutic use , Toxoplasmosis, Ocular/drug therapy , Adrenal Cortex Hormones/administration & dosage , Adult , Anti-Bacterial Agents/administration & dosage , Drug Therapy, Combination , Female , Humans , Male , Recurrence , Retrospective StudiesABSTRACT
BACKGROUND: Therapeutic regimens for the treatment of toxoplasmosis are not standardized. Treatment strategy mainly at the end of the second and the beginning of the third trimester, especially in cases of negative prenatal diagnosis, is the least uniform. In some situations, the choice of treatment may be ambiguous, and adverse drug reactions of the therapy should be taken into consideration. METHODS: Adverse drug reactions of anti-toxoplasma therapy with spiramycin (n = 77) versus pyrimethamine/sulfadiazine (n = 35) were compared in 112 pregnant women. RESULTS: Up to 36.6% of women reported adverse reactions to the treatment overall (n = 41). Out of those 38.9% (n = 30) were treated with spiramycin and 31.4% (n = 11) with pyrimethamine/sulfadiazine. Toxic allergic reactions were the only indication for discontinuation of treatment in 8.9% of patients (n = 10), where 9.1% (n = 7) were reported in spiramycin and 8.6% (n = 3) in pyrimethamine/sulfadiazine cohort. Neurotoxic complications (acral paraesthesia) were significantly more frequent during the therapy with spiramycine in 19.5% (n = 15) compared to no cases in pyrimethamine/sulfadiazine group (p = .003). Other adverse drug reactions, such as gastrointestinal discomfort, nephrotoxicity, vaginal discomfort were reported, but the differences between the cohorts were not significant. CONCLUSIONS: The superiority of one of the therapeutic regimens was not statistically demonstrated, since the differences in overall toxicity or incidence of toxic allergic reactions between the cohorts were not confirmed (p = .53 and p = 1.00, respectively). However, although the isolated neurotoxicity of spiramycin was the only significant adverse reaction demonstrated in this study, pyrimethamine/sulfadiazine therapy should be preferred, because it is known to be more effective and with limited adverse reactions.
Subject(s)
Drug-Related Side Effects and Adverse Reactions , Hypersensitivity , Spiramycin , Toxoplasmosis, Congenital , Toxoplasmosis , Female , Humans , Pregnancy , Spiramycin/adverse effects , Pyrimethamine/adverse effects , Sulfadiazine/adverse effects , Toxoplasmosis/drug therapy , Drug Therapy, Combination , Fetus , Hypersensitivity/drug therapy , Toxoplasmosis, Congenital/drug therapyABSTRACT
Pancreatic cancer (PDAC) has a poor prognosis despite surgical removal and adjuvant therapy. Additionally, the effects of postoperative analgesia with morphine and piritramide on survival among PDAC patients are unknown, as are their interactions with opioid/cannabinoid receptor gene expressions in PDAC tissue. Cancer-specific survival data for 71 PDAC patients who underwent radical surgery followed by postoperative analgesia with morphine (n = 48) or piritramide (n = 23) were therefore analyzed in conjunction with opioid/cannabinoid receptor gene expressions in the patients' tumors. Receptor gene expressions were determined using the quantitative real-time polymerase chain reaction. Patients receiving morphine had significantly longer cancer-specific survival (CSS) than those receiving piritramide postoperative analgesia (median 22.4 vs. 15 months; p = 0.038). This finding was supported by multivariate modelling (p < 0.001). The morphine and piritramide groups had similar morphine equipotent doses, receptor expression, and baseline characteristics. The opioid/cannabinoid receptor gene expression was analyzed in a group of 130 pancreatic cancer patients. Of the studied receptors, high cannabinoid receptor 2 (CB2) and opioid growth factor receptor (OGFR) gene expressions have a positive influence on the length of overall survival (OS; p = 0.029, resp. p = 0.01). Conversely, high delta opioid receptor gene expression shortened OS (p = 0.043). Multivariate modelling indicated that high CB2 and OGFR expression improved OS (HR = 0.538, p = 0.011, resp. HR = 0.435, p = 0.001), while high OPRD receptor expression shortened OS (HR = 2.264, p = 0.002). Morphine analgesia, CB2, and OGFR cancer tissue gene expression thus improved CSS resp. OS after radical PDAC surgery, whereas delta opioid receptor expression shortened OS.
ABSTRACT
Two weeks after delivery of a healthy term neonate, the mother developed lymph node syndrome, which corresponded serologically to acute toxoplasmosis. The blood of the newborn showed positive IgM, IgG and IgA antibody titers against Toxoplasma gondii with a low avidity of IgG. The newborn did not show any clinical signs or organ damage connected to toxoplasmosis either at the beginning or during the follow-up. The IgA and IgM titers exhibited a decrease over time, while the KFR, IgG antibody titers and avidity had an increasing trend. A sharp increase of KFR, IgE and IgA antibody titers was detected during the sixth month of life, probably due to maturation of the immune system in the setting of an asymptomatic infection with T. gondii. This short increase was followed by a subsequent decrease in titers of these antibodies until they reached negative levels during the 21st month of life. The evaluation of serological results in newborns infected with T. gondii is always difficult and should be performed by an expert physician. Children at risk should be placed under a long-term follow-up to avoid potential development of toxoplasma chorioretinitis.
Subject(s)
Toxoplasmosis, Congenital/diagnosis , Antibodies, Fungal/blood , Female , Humans , Infant , Infant, Newborn , Pregnancy , Toxoplasmosis/diagnosis , Toxoplasmosis/transmissionABSTRACT
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.
Subject(s)
Antibody Formation , COVID-19 , Antibodies, Viral , COVID-19/diagnosis , COVID-19/epidemiology , Czech Republic/epidemiology , Humans , Prospective Studies , SARS-CoV-2 , VaccinationABSTRACT
Background: Non-small cell lung cancer (NSCLC) is a leading cause of cancer-related death with a 5-year survival of only 21%. Reliable prognostic and/or predictive biomarkers are needed to improve NSCLC patient stratification, particularly in curative disease stages. Since the endogenous cannabinoid system is involved in both carcinogenesis and anticancer immune defense, we hypothesized that tumor tissue expression of cannabinoid 1 and 2 receptors (CB1 and CB2) may affect survival. Methods: Tumor tissue samples collected from 100 NSCLC patients undergoing radical surgery were analyzed for CB1 and CB2 gene and protein expression using the quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC). The gene and protein expression data were correlated with disease stage, histology, tumor grading, application of chemotherapy, and survival. Additional paired tumor and normal tissue samples of 10 NSCLC patients were analyzed independently for comparative analysis of CB1 and CB2 gene expression. Results: Patients with tumors expressing the CB2 gene had significantly longer overall survival (OS) (P<0.001), cancer specific survival (CSS) (P=0.002), and disease-free survival (DFS) (P<0.001). They also presented with fewer lymph node metastases at the time of surgery (P=0.011). A multivariate analysis identified CB2 tumor tissue gene expression as a positive prognostic factor for CSS [hazard ratio (HR) =0.274; P=0.013] and DFS (HR =0.322; P=0.009), and increased CSS. High CB2 gene and protein expression were detected in 79.6% and 31.5% of the tested tumor tissue samples, respectively. Neither CB1 gene nor CB1 or CB2 protein expression affected survival. When comparing paired tumor and tumor-free lung tissue samples, we observed reduced CB1 (P=0.008) and CB1 (P=0.056) gene expression in tumor tissues. Conclusions: In NSCLC patients undergoing radical surgery, expression of the CB1 and CB2 receptor genes is significantly decreased in neoplastic versus tumor-free lung tissue. CB2 tumor tissue gene expression is strongly associated with longer survival (OS, CSS, DFS) and fewer lymph node metastases at the time of surgery. More studies are needed to evaluate its role as a biomarker in NSCLC and to investigate the potential use of CB2 modulators to treat or prevent lung cancers.
ABSTRACT
OBJECTIVES: Recapitulation of our knowledge dealing with diagnosis and therapy of oral mucosal diseases caused by HSV. MATERIAL AND METHODS: Analysis of a sample of 135 patients coming from Department of Dentistry, Faculty of Medicine in Hradec Králové within 11 years. Statistical evaluation of the data and related associations (clinical diagnosis, age, gender, general health status, treatment mode) were realized by ANOVA analysis, t-test, and Fisher's exact test. RESULTS: A total sample of 135 individuals (81 women and 54 men) consisted of 49 individuals with primary herpetic gingivostomatitis (37 %) and recurrences with clinical signs of herpetic stomatitis in 68 patients (50 %) and labial herpes simplex in 18 patients (13 %) aged from 1 to 90 years, mean age 40.4 year, with maximal occurrence in the 3rd and 7th decades. The mean age of individuals with herpetic gingivostomatitis was significantly lower (p = 0.0001). The occurrence of herpetic stomatitis was associated with severe concomitant diseases (p = 0,0001). Topical treatment was applied in 73 individuals (54 %), combined mode in 58 individuals (43 %), only systemic therapy in 4 individuals (3 %). In most of individuals with herpetic gingivostomatitis only topical treatment was used (80 %, in most of herpetic recurrences combined therapy was applied (62 %). CONCLUSIONS: The age of individuals with primary infection and herpetic recurrences was significantly different. Herpetic stomatitis occurred often as an opportunistic viral infection in seriously ill and older patients. Antiseptics with antiviral properties and their combinations with systemic administration of antiviral drugs proved successfully in the treatment.
Subject(s)
Herpes Labialis , Stomatitis, Herpetic , Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Female , Herpes Labialis/diagnosis , Herpes Labialis/therapy , Humans , Infant , Male , Middle Aged , Stomatitis, Herpetic/diagnosis , Stomatitis, Herpetic/therapy , Young AdultABSTRACT
Infections caused by pathogenic protozoan Toxoplasma gondii in our geographic area is the most frequent parasitic infection; Czech Republic declares seroprevalence approx. 30 %. Diagnosis of toxoplasmosis is mostly based on serological methods are used (EIA IgM, IgA, IgE, IgG and avidity in IgG, Western Blot, complement fixation). According to positive results of these tests diagnosis of acute toxoplasmosis is established. In our retrospective study we tried to evaluate results of T. gondii DNA positivity from blood samples by PCR compared with positive markers of acute infection in patients before specific therapy was initiated. In accordance with literature we concluded, that in routine examination of immunocompetent outpatients of Clinic of Infectious Diseases from the moment of 4 weeks after lymphatic nodes swelling protozoan DNA detection in blood sample is not possible.
Subject(s)
DNA, Protozoan/blood , Polymerase Chain Reaction , Toxoplasmosis/diagnosis , Adolescent , Adult , Child , Child, Preschool , Complement Fixation Tests , Female , Humans , Immunocompetence , Immunoglobulins/blood , Male , Middle Aged , Toxoplasma/genetics , Toxoplasma/isolation & purification , Toxoplasmosis/immunology , Young AdultABSTRACT
Since introduction of polymerase chain reaction for the detection of DNA Toxoplasma gondii 398 biological samples from 301 patients were examined from 2000 to 2003. Positive finding of toxoplasmosis DNA we noted in 23 cases. Polymerase chain reaction enables exact and fast diagnosis of the actual parasitemia Toxoplasma gondii, which is important especially in the pregnant patients, new-born with suspicion on congenital toxoplasmosis and in the patients with immunosupression.
Subject(s)
DNA, Protozoan/analysis , Polymerase Chain Reaction , Toxoplasmosis/diagnosis , Animals , Female , Humans , Pregnancy , Toxoplasma/genetics , Toxoplasmosis, Congenital/diagnosis , Toxoplasmosis, Ocular/diagnosisABSTRACT
Coinfection by HIV and syphilis has become a growing problem due to the re-appearance of unsafe sexual practices in the era of potent anti-retroviral drugs. We describe a repeated import of syphilis by a couple of men-who-have-sex-with-men from Thailand to Czech Republic likely due to non-adherence of the patients to physician recommendations. Such cases can become foci for dissemination of once locally rare infections and present a danger for the community.
Subject(s)
HIV Infections/microbiology , Homosexuality, Male , Syphilis/virology , Coinfection , Czech Republic , HIV Infections/ethnology , HIV Infections/immunology , Humans , Male , Patient Compliance , Prevalence , Recurrence , Syphilis/ethnology , Syphilis/immunology , Syphilis/transmission , Thailand/ethnologyABSTRACT
Case of 59-year-old male with chronic obstructive pulmonary disease and a number of comorbidities, who has developed meningoencephalitis caused by Cryptococcus neoformans var. grubii with polyarteritis nodosa diagnosed during hospitalization, was presented. Before evidence of meningoencephalitis, the patient was being treated with ketoconazole and low doses of fluconazole (200 mg/day) for alleged candidiasis. The dosage was increased (800 mg/day) following laboratory diagnosis of C. neoformans based on positive latex agglutination test and biochemical identification of encapsulated yeast isolated from the blood and CSF. Later, the yeast identification was confirmed by sequencing analysis. Owing to inadequate clinical response, fluconazole therapy was switched to voriconazole (400 mg/day) and later to intravenous amphotericin B (1.0 mg/kg per day). Despite of a temporary stabilization and improvement, which correlated with decline of cryptococcal antigen titers (from 1:1024 to 1:8), after 6 weeks, the patient's underlying condition deteriorated due to severe pancolitis and serious nosocomial bacterial infections. The patient died of multiorgan failure several days later. Our case demonstrates a possible connection between the development of life-threatening cryptococcosis and an autoimmune vasculitis disease and emphasizes that the outcome of the management of cryptococcal meningoencephalitis is highly dependent on early diagnosis, adequate treatment, including dosage, and last but not least control of underlying disease and risk factors.
Subject(s)
Cryptococcus neoformans/isolation & purification , Meningoencephalitis/microbiology , Polyarteritis Nodosa/complications , Antifungal Agents/administration & dosage , Cryptococcus neoformans/drug effects , Cryptococcus neoformans/genetics , Fatal Outcome , Fluconazole/administration & dosage , Humans , Male , Meningoencephalitis/drug therapy , Meningoencephalitis/etiology , Middle Aged , Polyarteritis Nodosa/microbiologySubject(s)
Chickenpox/diagnosis , Cystic Fibrosis/complications , Herpesvirus 3, Human/isolation & purification , Skin/pathology , Chickenpox/immunology , Child , DNA, Viral/genetics , DNA, Viral/isolation & purification , Female , Genotype , Herpesvirus 3, Human/classification , Herpesvirus 3, Human/genetics , Humans , Skin/virologyABSTRACT
Varicella zoster virus typically causes a benign disease in childhood called varicella (chickenpox) and can reactivate in adults as a dermatomally distributed, painful rash illness known as herpes zoster (HZ). Infection with VZV can however lead to severe complications in immunocompromised patients that can result in hospitalization and, occasionally, death. Here we describe a patient, who acquired primary VZV infection during a 3-week-long treatment regimen with corticosteroids. The disease took a fulminant course, leading to a liver failure and severe coagulopathy. The patient died 9 days following hospital admission, despite intensive antiviral and supportive treatment. Wild-type VZV DNA was detected from multiple samples from esophagus, liver and skin. Genotypic analysis based on single nucleotide polymorphism profiles in open reading frames (ORFs) 21, 22 and 50 identified this strain as a clade 4 isolate, which is typically found in tropical countries. This is the first description of a clade 4 strain from a patient in the Czech Republic.
Subject(s)
Chickenpox/complications , Hepatitis/complications , Herpesvirus 3, Human/physiology , Immunocompromised Host , Adult , Czech Republic , Fatal Outcome , Female , Genes, Viral/genetics , Genotype , Hepatitis/pathology , Herpesvirus 3, Human/classification , Herpesvirus 3, Human/genetics , Humans , Immunosuppressive Agents/adverse effectsABSTRACT
OBJECTIVES: Monitoring of the varicella-zoster virus is becoming an important tool for analysis of the circulation of individual strains of VZV which differ not only at the genomic level, but show a variability in their clinical and epidemiological characteristics. Such data are not available on a large scale from the Czech population and could help understanding the epidemiological and evolutionary characteristics of the virus, as well as its potential for reinfection and increased pathogenesis in the population groups at higher risk for complications. The main aim of this study was detection and monitoring of wild-type or vaccine VZV strain isolates in the region of Eastern Bohemia and genotypic characterization of these isolates. MATERIAL: A total of 273 clinical samples were obtained from patients exhibiting symptoms of varicella zoster virus (VZV) infection manifested as chickenpox or herpes zoster (HZ) treated in the Faculty Hospital of Charles University, Medical School in Hradec Kralove, Czech Republic. METHODS: Characterization of individual short VZV DNA sequences was performed utilizing restriction fragment length polymorphisms (RFLPs), PCR and sequencing. Single nucleotide polymorphisms (SNP) in open reading frames (ORF) 21, 22 and 50 were used to identify individual VZV strains. RESULTS: All clinical isolates (97 from varicella, 176 from herpes zoster) were VZV positive wild-type strains. Sequencing analysis showed that 89 isolates were of the European E1 genotype, 180 were of the European E2 genotype and 2 were identified as the Mosaic M1 strain. In addition, for the first time in this region two unusual genotypes were identified, both representing a combination of E1 and M2 strain specific SNPs. CONCLUSION: Our prospective VZV genotyping study which is the first to monitor the VZV epidemiological situation in the Czech Republic using such a large set of clinical specimens, has provided valuable epidemiological data and identified two unique VZV recombinants.
Subject(s)
Genotype , Herpesvirus 3, Human/genetics , Adolescent , Adult , Aged , Aged, 80 and over , Chickenpox/virology , Child , Child, Preschool , Czech Republic , DNA, Viral/genetics , Female , Herpes Zoster/virology , Herpesvirus 3, Human/isolation & purification , Humans , Male , Middle Aged , Open Reading Frames , Polymorphism, Restriction Fragment Length , Polymorphism, Single Nucleotide , Young AdultABSTRACT
AIMS: We report a case of multi-recurrent herpes zoster in a 53-year-old Caucasian woman treated repeatedly at the Faculty Hospital Hradec Kralove, Czech Republic over the years 2009 - 2011. METHODS: Specific PCR methods targeting single nucleotide polymorphisms (SNPs) in open reading frames (ORF) 38, 54 and 62 were utilized to determine vaccine or wild type varicella-zoster (VZV) strains followed by SNPs analysis using two amplicons in ORF 22 and/or ORF 21/ORF 50. Additional genotyping in ORF 1, 6, 9 and 28 was subsequently performed due to the unusual results. RESULTS: Three sets of clinical specimens from one patient (from hospital visits 2, 3 and 4) were analyzed and the presence of an unusual wild-type strain of VZV was discovered. The VZV strain isolated from the lesions bears a combination of markers characteristic both for Mosaic 2 (M2) and European 1 (E1) wild-type VZV strains. CONCLUSION: This is the first report of atypical wild-type VZV strain circulating currently in Czech Republic.