[Assessment of body mass index in patients with chronic hepatitis C who received an effective antiviral therapy].

AIM: To evaluate the body mass index (BMI) in patients with chronic hepatitis C (CHC) with different stages of liver fibrosis and steatosis who received effective antiviral therapy (AVT). MATERIALS AND METHODS: The study included 278 CHC patients with a sustained virologic response (SVR) at the end of treatment. In addition to assessing the investigational data to determine the clinical status of the patient, we calculated BMI (following the World Health Organization guidelines) and determined the severity of liver fibrosis (F) and steatosis (S) using transient elastography. The patients were assessed at the start of antiviral therapy, after ≥6 months from the moment SVR was confirmed, and then every 12 to 24 months. RESULTS: By the end of the study, the mean patient age was 49 years, 53% of them were men, and 34% of the patients were obese. Excessive weight gain was registered in 17% (n=48) of the cases, with 60% newly diagnosed with Class 1 to 2 obesity. Both before the start of AVT and years after reaching SVR, the mean BMI corresponded to the reference pre-obesity values, the liver steatosis was significantly more often absent in normal BMI; on the contrary, fatty liver (predominantly S2 to S3) was registered in individuals with elevated BMI (p<0.0001). After the long-term period following a successful therapy, Stage F4 liver fibrosis patients were mainly diagnosed with obesity (80% versus 44% before AVT; p=0.0010). CONCLUSION: The high proportion of patients with elevated BMI and liver steatosis seen years after a successful CHC therapy indicates a continued risk of progression of chronic liver disease. Such patients should be advised on how important it is to change their lifestyle to reduce overweight and prevent weight gain. We also need long-term assessments of how liver steatosis changes over time and what are the outcomes associated with post-SVR increase in BMI.

Decreased plasma levels of 25(OH)D in multiple sclerosis patients. Correlation with disease severity expressed by EDSS, MSSS, progression index and Herbert´s scale severity grade.

OBJECTIVES: Multiple sclerosis is a chronic inflammatory and autoimmune demyelinating disease of the brain and spinal cord. Vitamin D has anti-inflammatory and anti-Th1, Th17 activities, activates the function of regulatory T cells, shifts the immune response towards Th2, so it might be favorable for downregulation of the disease pathogenesis, and if inflammation and Th1 and Th17 immunity are hyperactivated. The aim of our study was to highlight the role of vitamin D in multiple sclerosis pathogenesis. METHODS: We investigated 178 patients with multiple sclerosis. Plasma levels of 25(OH)D and HMGB1 were investigated. RESULTS: Despite a regular use of VD by patients, the plasma levels of 25(0H)D were significantly decreased in 57% of them, 14.1% had VD deficiency (level of 25(OH)D < 20 ng/mL) and more than 6 % of patients had VD severe deficiency with the plasma level of 25(OH)D < 12 ng/mL. The level of 25(OH)D negatively correlated with the severity of the disease (EDSS, index of progression, duration of the disease) and negative association was found also with Herbert´s six severity grades. HMGB1 levels were higher in patients (p < 0.0001). CONCLUSION: Our result showed that vitamin D deficiency plays a role in multiple sclerosis pathogenesis. We believe that administration of vitamin D to patients at a sufficient dose providing a physiological level of vitamin D could have a positive effect on the course of the disease. However, regular monitoring of vitamin D levels is required, which should be at least within 30-75 ng/mL, and even more, but below the toxicity limit (Tab. 6, Ref. 66).

[Liver-induced fibrosis in young and old animals is accompanied by age-dependent changes in bone marrow cells.]

The hypothesis of dependence of the functional characterizations of bone marrow cells (BMC) - proliferation rate, direction proliferation, etc. - not only by the age of animals, but also on features of BMC microenvironment was verified. Two methods of changing the microenvironment were used. There were in vivo (induction of liver fibrosis in young and old animals) and in vitro (transfer of young and old animals BMC obtained in intact animals and animals with fibrosis into the same standard culture system). CuSO4-induced liver fibrosis and CCl4-induced liver fibrosis had a different effect on the ratio of cell types in the bone marrow in young and old animals. Thus, in young animals, regardless of the type of liver fibrosis inducer, the relative number of morphologically identifiable cell types decreased. This was accompanied by an increase in the number of identified cell types against the background of CuSO4-induced liver fibrosis and did not change against the background of CCl4-induced liver fibrosis in old animals. The proliferative activity of BMC isolated from old animals and transferred to an in vitro culture was superior to that of young animals. This is due to the large number of lymphocytes in the bone marrow of old animals by 167% and the specific composition and characteristics of the BMC microenvironment in old animals.

[Efficacy and safety of narlaprevir/ritonavir and daclatasvir non interferon combination in population of Russian patients with chronic hepatitis C].

AIM: Evaluate efficacy and safety of a combination of direct - acting antivirals narlaprevir/ritonavir with daclatasvir in patients with viral hepatitis C. MATERIALS AND METHODS: The study enrolled adult patients with HCV genotype 1b infection without demonstrated NS5A resistance - associated substitutions Y93C/H/N/S and/or L31F/M/V/I. Patients were treated with narlaprevir 200 mg QD, ritonavir 100 mg QD and daclatasvir 60 mg QD. Treatment duration was 12 weeks. Proportion of patients achieving sustained virological response 12 weeks after treatment (SVR12) was the primary efficacy endpoint. RESULTS AND DISCUSSION: In total, 105 (75.0%) patients were treatment with the study combination. Patients' age varied from 21 to 69 years, the mean age being 43.2±10.9 years. There were slightly more women (55.2%), and 69 patients (65.7%) had comorbidities. SVR 12 was 89.5% (95% CI 82.0-94.7%). In 10 of 11 patients with treatment failures NS5A resistance - associated substitutions in residues 31 and/or 93 were found, as well as less clinically relevant substitutions L28M, P58S, R30Q, Q62K. Adverse events (AEs) were found in less than one half of patients (45 patients, or 42.9% in the safety population). Almost all recorded AEs were mild to moderate. CONCLUSION: Efficacy of treatment with a combination of narlaprevir/ritonavir and daclatasvir in treatment - naïve patients with HCV genotype 1b was close to 90%. This combination was found to be safe and well - tolerated.

Inhibitory effect of streptococci on the growth of M. catarrhalis strains and the diversity of putative bacteriocin-like gene loci in the genomes of S. pneumoniae and its relatives.

S. pneumoniae is a facultative human pathogen causing a wide range of infections including the life-threatening pneumoniae or meningitis. It colonizes nasopharynx as well as its closest phylogenetic relatives S. pseudopneumoniae and S. mitis. Both the latter, despite the considerable morphological and phenotypic similarity with the pneumococcus, are considerably less pathogenic for humans and cause infections mainly in the immunocompromized hosts. In this work, we compared the inhibitory effect of S. pneumoniae and its relatives on the growth of Moraxella catarrhalis strains using the culture-based antagonistic test. We observed that the inhibitory effect of S. mitis strains is kept when a hydrogen peroxide produced by cells is inactivated by catalase, and even when the live cells are killed in chloroform vapors, in contrast to the pneumococcus whose inhibiting ability disappeared when the cells die. It was suggested that this effect may be due to the production of bacterial antimicrobial peptides by S. mitis, so we examined the genomes of our strains for the presence of bacteriocin-like peptides encoding genes. We observed that a set of bacteriocin-like genes in the genome of S. mitis is greatly poorer in comparison with S. pneumoniae one; moreover, in one S. mitis strain we found no bacteriocin-like genes. It could mean that there are probably some additional opportunities of S. mitis to inhibit the growth of competing neighbors which are still have to be discovered.

Mediators and Biomarkers of Inflammation in Meningitis: Cytokine and Peptidome Profiling of Cerebrospinal Fluid.

Differential diagnosis of bacterial and viral meningitis is an urgent problem of the modern clinical medicine. Early and accurate detection of meningitis etiology largely determines the strategy of its treatment and significantly increases the likelihood of a favorable outcome for the patient. In the present work, we analyzed the peptidome and cytokine profiles of cerebrospinal fluid (CSF) of 17 patients with meningitis of bacterial and viral etiology and of 20 neurologically healthy controls. In addition to the identified peptides (potential biomarkers), we found significant differences in the cytokine status of the CSF of the patients. We found that cut-off of 100 pg/ml of IL-1ß, TNF, and GM-CSF levels discriminates bacterial and viral meningitis with 100% specificity and selectivity. We demonstrated for the first time the reduction in the level of two cytokines, IL-13 and GM-CSF, in the CSF of patients with viral meningitis in comparison with the controls. The decrease in GM-CSF level in the CSF of patients with viral meningitis can be explained by a disproportionate increase in the levels of cytokines IL-10, IFN-γ, and IL-4, which inhibit the GM-CSF expression, whereas IL-1, IL-6, and TNF activate it. These observations suggest an additional approach for differential diagnosis of bacterial and viral meningitis based on the normalized ratio IL-10/IL-1ß and IL-10/TNF > 1, as well as on the ratio IFN-γ/IL-1ß and IFN-γ/TNF < 0.1. Our findings extend the panel of promising clinical and diagnostic biomarkers of viral and bacterial meningitis and reveal opposite changes in the cytokine expression in meningitis due to compensatory action of pro- and antiinflammatory factors.

[The effect of cytokine therapy on the results of surgical treatment of patients with acute ulcerative gastroduodenal bleeding].

AIM: To improve the results of surgical treatment of patients with acute ulcerative gastroduodenal bleeding using immune correction with ß-leukin. MATERIAL AND METHODS: Urgent and elective operations were performed in 120 patients with gastroduodenal ulcers. Patients of the main group (n=63) received systemic cytokine therapy with ß-leukin in addition to conventional treatment. Pre- and postoperative immune correction was absent in the comparison group. RESULTS AND DISCUSSION: There was preoperative imbalance of antiphlogistic and proinflammatory cytokines in comparison group. Severity of this process depended on blood loss. Therapy with interleukin decreased severity of cytokine imbalance.

[Possibilities of using cepeginterferon alpha-2b in double (cepeginterferon alfa-2b and ribavirin) and triple (simeprevir, cepeginterferon alpha-2b, and ribavirin) antiviral therapy regimens for chronic hepatitis C. A review of clinical trials and experience of everyday clinical practice]. / Vozmozhnosti primeneniia tsepeginterferona al'fa-2b v sostave dvukh- (tsepeginterferon al'fa-2b i ribavirin) i trekhkomponentnoi (simeprevir, tsepeginterferon al'fa-2b i ribavirin) skhem protivovirusnoi terapii khronicheskogo gepatita S. Obzor klinicheskikh issledovanii i opyta povsednevnoi klinicheskoi praktiki.

Since the incidence of chronic hepatitis C (CHC) increases steadily, the priority of national health care is to provide antiviral therapy (AVT) for the maximum number of patients infected with hepatitis C virus (HCV). The regimens including pegylated interferons (PEG-IFN) are still in demand in the Russian Federation. A number of clinical trials have been conducted to evaluate the efficacy and safety of cepeginterferon alpha-2b (cePEG-IFN alpha-2b), an original PEG-IFN-α developed in the Russian Federation. Their results have shown that cePEG-IFN alpha-2b in the two-component AVT regimen has at least no less clinical efficacy than PEG-IFN alpha-2b and PEG-INF alpha-2a in HCV monoinfected and HCV/HIV co-infected patients. The pooled analysis of data has indicated that the use of cePEG-IFN alpha-b in combination with ribavirin allows an average of 80% of the patients with HCV genotypes 2 and 3 and 62% of those with HCV genotype 1 to achieve a sustained virological response (SVR). In clinical practice when the two-component AVT regimen (cePEG-IFN alpha-b and ribavirin) was used in patients with early-stage CHC and mild fibrosis, SVR was recorded in 90.7% of the patients with HCV genotype 2/3 and in 75% of those with HCV genotype 1. The experience in using cePEG-IFN alpha-2b as a component of the three-component AVT regimen (simeprevir, cePEG IFN alfa-2b, and ribavirin) has been published. The observational program manly covered young patients with mild or moderate fibrosis. SVR was observed in 94% of the patients. Another paper describes the experience with the triple AVT therapy (simeprevir, cePEG-IFN alfa-2b, and ribavirin) in 22 patients, the majority of whom had advanced fibrosis. SVR was recorded in 71.4% of those who had completed treatment. Thus, an individual approach and assessment of predictive response factors to two- or three-component AVT regimens including cePEG-IFN alpha 2b can achieve successful treatment outcomes in most patients with CHC, which is, in some cases, more economically sound than interferon-free regimens used as first-line therapy.

Young and old animals use various strategies of metabolic adaptation to Cu-induced liver fibrosis.

The role of the prooxidant-antioxidant and immune systems in formation of age-related adaptive responses to chronic effects of toxic substances (copper ions) was studied. In this purpose in young and old animals by multiple sequential administration of copper sulfate was induced liver fibrosis. Using this model the characteristics of prooxidant-antioxidant and immune systems, and physiological characteristics of were assessed. The Cu-induced liver fibrosis was accompanied by the oxidative stress and a significant reduction in cell immune activity. In the case of elimination of oxidative stress by introducing exogenous antioxidants (mix factor) in animals with fibrosis was recovered efficiency, body weight, the content of red blood cells. In old animals the restoring of the changed parameters of cellular immunity was more effective than in the young. Young and old animals used different metabolic strategies of adaptation to toxic environmental factors.

[State of immune and cytokine status in acute ulcerative gastroduodenal bleeding].

The paper analyzes the results of a survey of 37 patients with acute ulcerative gastroduodenal hemorrhage. During endoscopy found that 7 patients the cause of bleeding was gastric ulcer, duodenal ulcer in 27 and 3 concomitant ulcer. Hemostasis was evaluated by J. Forrest and the severity of blood loss was assessed by the A.A. Shalimov: with blood loss was mild in 11 patients, the average degree--12 and with severe--14. In all patients on admission in the peripheral blood were studied state of cellular, humoral and cytokine profile: CD3+, CD4+, CD8+, CD4+/CD8 + and CD19+, FI, FF, CEC, IgA, M, G, TNF-α, IFN-γ, IL-1, IL-2, IL-4, IL-6, IL-8 and IL-10. In general patients with on admission revealed significant changes in the immune and cytokine status. In cellular immunity occurs immunosuppression. Disturbances in humoral immunity manifests itself in increased levels of lymphocytes in blood loss and hardships of all multi-directional changes in the concentration of immunoglobulins depending on the amount of blood loss. When there is an imbalance in cytokine status, the depth of which depends on the degree of blood loss.

[Effect of combined and local cytokine- and ozone therapy on the indices of lipid peroxidation, endogenous intoxication and ferroproteins in diffuse peritonitis].

The article analyzes the results of effect of combined and local cytokine- and ozone therapy on the indices of lipid peroxidation, endogenous intoxication and ferroproteins in 111 patients with diffuse peritonitis. It was shown, that combined sequential local and systemic cytokine and ozone therapy allows correcting the expression of endogenous intoxication and lipid peroxidation in diffuse peritonitis. This method suppresses an inflammation in the abdominal cavity. At the same time, it accelerates the elimination of intestine atony and thereby potentiates the possibilities of traditional methods of treatment.

[Comparative estimation of efficacy of the immunecorrecting combined three- level cytokino- and ozonotherapy in complex treatment of extended peritonitis].

The results of surgical treatment of 211 patients, suffering extended peritonitis (EP) of various etiology, were analyzed. The peritonitis severity was graded in accordance to Mannheim's index of peritonitis (MIP). The patients were divided on two groups. In 60 patients (group of comparison) the basic treatment was conducted, without immunocorrection. The patients of the main group were divided on three subgroups. To the patients of the first subgroup (43) 400 ml of ozonated isotonic solution (OIS) of sodium chloride was infused intravenously additionally in the contents of basic therapy, as well as peritoneal-enteral detoxication using OIS was conducted. In a second subgroup a regional intraabdominal endolymphatic ozonotherapy (OTH) was conducted to 57 patients with OIS and peritoneal-enteral sanation using medical ozone. In the 3d subgroup in 51 patients additionally were applied intravenous infusion of OIS and peritoneal-enteral sanation with medical ozone. To these patients cytokinotherapy was conducted, when splenopid was applied intravenously, intraperitoneally and enterally simultaneously. The indices of the T- and B-immunity links, phagocytic activity of neutrophils, including phagocytic index, phagocytic number, the completeness of phagocytosis index, as well as the content of the tumor necrosis factor alpha (TNF-alpha), gamma-interferon (IFN-gamma) and interleukins (IL) - IL-1, IL-2, IL-4, IL-6, IL-10 were estimated in the blood serum in dynamics. The combined staged three-level (systemic, intraperitoneal and enteral) application of natural cytokins and medical ozone have promoted mutual potentiating of their action, significant efficacy of the immunemodulating therapy in comparison with such systemic and local OTH.

[Immunogenetic predictors of a rapid virologic response to antiviral therapy in patients with chronic hepatitis C].

AIM: To determine the correlation between interleukin 28B (IL28B) gene polymorphism in patients with chronic hepatitis C (CHC), the presence or absence a rapid virologic response to antiviral therapy, and a number of immunological characteristics as a basis for a personalized approach to treating the patients. SUBJECTS AND METHODS: Seventeen CHC patients infected with hepatitis C virus genotype 1b were examined and underwent genetic testing for IL28B gene polymorphism for rs12979860 (CC, CT or TT genotypes) and rs8099917 (TT, TG or GG genotypes) using the modified method of adjacent samples, which revealed single nucleotide substitutions in the genes. Their immunological parameters were identified by a flow cytometry technique by taking into account whether a rapid virologic response had been achieved. RESULTS: The key phenomena of a rapid virologic response in the representatives of different IL28B genotypes are the nonspecific proliferative activity of blood natural killer cells before treatment, as well as the count of regulatory T cells before and 4 weeks after therapy start. CONCLUSION: To predict the efficiency of antiviral therapy for CHC, it is desirable to supplement genetic studies with immunological data.

[The influence of ozone therapy to some immunological parameters in patients with generalized peritonitis].

The immune status of the surgical treatment of 103 patients with generalized peritonitis (GP) were discussed. Patients were divided into two groups. Comparison group included 60 patients who received conventional combined therapy, main group - 43 patients, who, along with the standard complex treatment measures, were used local and systemic intravenous ozone therapy (OT). The severity state of patients evaluated by Mannheim peritoneal index (MPI). To assess the immune status in blood of the patients with GP were studied the level of lymphocyte subpopulations (CD3+, CD4+, CD8+, CD11a+, CD19+), the phagocytic activity of neutrophilic granulocytes, concentration of immunoglobulin A, M, G, and the content of circulating immune complexes (CIC). Studies have shown that GP is accompanied by changes in the functioning of the immune system, the severity and extent of which depends on the severity of MPI. The conventional basic therapy is not effective in correcting of these disturbances. Using the local and systemic OT in the complex of therapeutic measures in individually selected patients with GP have a positive effect on some level of immunity with a tendency to restore the immune system dysfunction.

[K+/Na+ in the animal extracellular fluid at weathering of granitoids and problem of the origin of life].

Leaching of granitoids of the paleoproterozoic age was performed from several seconds to 360 days in water solutions (pH < 6.0) in the oxygen or argon medium. Under these conditions the entrance of K+ into the solution has been shown to occur at the higher rate than that of Na+. The obtained data are considered as evidence in favor of a possibility of the existence at the earlier stages of the Earth geologic history of water reservoirs with predominance of K+ over Na+. The K+/Na+ ratio exceeding 1 has been shown in prokaryotes, in cells and tissues of the free living and parasitic species of invertebrate and vertebrate animals. At the same time in the extracellular fluid of the fresh water, marine, and terrestrial animals, in which the Na+ concentration varies from 13 to 482 mmol/l, the K+/Na+ ratio is preserved at the level of 0.034 +/- 0.001. These results are discussed in connection with the problem of origin of protocells and of concentration ratios of monovalent cations in water phases of multicellular organisms.

[Using the operonic gene pairs for establishing the treshold for correlation coefficient of differently expressed genes].

We developed an approach for effective estimating the correlations in the noise component of gene expression data. An efficent noise reduction technique has been suggested. The resulting technique has been applied to E. coli microarray data and tested on SOS response modulated genes.

[Outcomes of severe pandemic influenza A/H1N1/2009].

AIM: To study the outcomes of severe pandemic influenza A/H1N1/2009. SUBJECTS AND METHODS: The study enrolled 24 patients, including 8 males and 16 females (10 of whom were pregnant), aged 17 to 58 years, with a laboratorily verified diagnosis of pandemic influenza A/H1N1/2009, treated at the intensive care unit for the significant symptoms of acute respiratory failure (RF). Real-time RT-PCR was used to verify the diagnosis. Organs and tissues from deceased patients were histologically studied; chest computed tomography, body plethysmography, fibrobronchoscopy, breath test, and 6-minute walk test were performed in the late period. RESULTS: Within the first 30 days, a fatal outcome caused by therapy-resistant progressive RF was observed in 33% of the patients with pandemic influenza treated at the intensive care unit. Diffuse alveolar damage caused by influenza virus, which gives rise to hyaline membranes, underlies RF. Lung tissue fibrosis formed in recovered patients. CONCLUSION: The severity of pandemic influenza A/H1N1/2009 was determined by massive bilateral pneumonia, interstitial (alveolar) pulmonary edema, formation of diffuse bilateral lung fibrosis at the outcome of severe virus pneumonia (acute respiratory distress syndrome) with a decrease in vital and diffusing capacities, thereby generating a need to follow up this patient category and, possibly, to elaborate special rehabilitation programs.

[Supposed role of "metabolic memory" in formation of response reaction to stress-factors in young and adult organisms].

The influence of the combined long-lasted influences of sulfur sulfate and diet restriction in young (3 month age) and adult (21 month age) Vistar rats on activity of glucose-6-phospatase, alaninaminotranspherase (ALT), aspartataminotranspherase (AST), and on phosphorilating activity of liver mitochondria was studied to investigate the role of metabolic memory on the peculiarities of response reaction. The young animals not differed from adult ones in the possibility of inducing activity of glucose-6-phospatase, ALT, and on phosphorilating activity after the influence of sulfur sulfate and diet restriction. The age-related differences in glucose-6-phospatase and transpherases and phosphorilating activity existing in control disappeared after the long-lasted action of sulfur sulfate and diet restriction. The answer reaction in enzyme activity to stress factors applied many times depends upon the metabolic memory formed in the process of adaptation, and the age of animals have no influence on it. In some relation the ontogenesis may be considered as a result of adaptation genesis. The metabolic memory can change the answer of the system to the stress influence. There are three types of modification of the answer to stress factors: the answer remains unchanged (metabolic memory), "paradox answer" formation, and super activation of the metabolic system.

[HCV genome variability in acute and chronic viral hepatitis C].

AIM: To evaluate HCV genome variability in acute and chronic phases of viral hepatitis C. MATERIAL AND METHODS: The study of heterogeneity of HCV in acute hepatitis C has detected genetic heterogeneity and variability of individual HCV population circulating in the blood. Significant genetic heterogeneity of HCV was observed in 1b, 2a and 3a genotypes. Variability of HCV did not depend on virus load. Genetic HCV structure changed significantly both in patients with manifest ALT deviations and in normal ALT, mean number of HCV genetic variants in these groups being the same. No significant correlations were found between virus concentration in the patient's blood, its variability and ALT values. Genetic heterogeneity of interferon-sensitive region of gene NS5A subtype 1b HCV was studied in blood of 16 patients with chronic hepatitis C resistant to interferon therapy. RESULTS: It is shown that genetic heterogeneity and variability of an individual HCV population circulating in blood serum can not be a prognostic criterion in assessment of variants of acute hepatitis C course. No mutations in ISDR region were found in 25% of 16 patients studied. 75% cases had 1-3 replacements of amino acid sequences, most frequent mutation was replacements in position 2218 (histidin/arginin). The above results are close to those obtained in Japanese and European populations. Results of ISDR sequence-analysis conducted before treatment may predict efficacy of interferon-alpha2 treatment in an individual patient in future. Large-scale trials are necessary for detection of mutations responsible for resistance to interferon-alpha2 in patients living in Russia.

Hepatitis B virus genetic typing using mass-spectrometry.

Mini-sequencing with subsequent result registration using MALDI-ToF mass-spectrometry was employed for hepatitis B virus genetic typing in Russian population. This approach was employed for hepatitis B virus genetic typing in HBsAg-positive patients with chronic hepatitis B, hepatitis of combined etiology and hepatic cirrhosis and allowed to show the prevalence of D genotype (83.3%) in all groups of patients. Other hepatitis B virus genotypes: genotype A (5.9%), genotype C (3.6%), and mixed infection with D and C (7.2%) were also found in patients with chronic hepatitis B and hepatic cirrhosis. All genotypes were found in patients with chronic hepatitis B and hepatic cirrhosis. Chronic hepatitis of combined etiology was noted only in patients with genotype D. Possibility of detection of mixed infection with hepatitis B viruses of various genotypes is a distinct advantage of mini-sequencing approach over direct nucleotide sequence evaluation for hepatitis B virus genetic typing.