Significance of genotyping in the diagnosis of arthritis associated with C-viral hepatitis

To date, a number of studies have been conducted on the relationship of HCV genotypes with complications such as liver cirrhosis, hepatocellular carcinoma, but research on the correlation of the genotype with extrahepatic clinical manifestations associated with HCV, especially rheumatic manifestations, is insufficient. The purpose of the study is to analyze the most common genotypes of viral hepatitis C in the Republic of Uzbekistan and study their correlation with rheumatic manifestations of HCV. Methods: This study involved 88 patients with HCV infection who received inpatient and subsequent ambulatornoe treatment in Scientific Research Institute of Epidemiology, Microbiology and Infectious diseases of the Republic of Uzbekistan. Diagnostics procedures at the initial level included a set of studies, i. e. clinical, rheumatological and laboratory studies, ultrasound of the joint. In addition, 88 samples of RNA - positive serum from patients diagnosed with HCV were genotyped. Results: the Results showed that genotype 1 of HCV (subtype 1b) is the main genetic variant of HCV in Uzbekistan. Based on the results obtained, the genotypic features of viruses can be markers of the development of extrahepatic clinical manifestations. There is a reliable direct correlation between the incidence and severity of associated arthritis in patients with chronic viral hepatitis C with genotype 1b of the virus according to our research. Conclusions: The results of the study can be provided to help the doctor of the polyclinic service as recommendations that patients with the 1b genotype of the virus have a greater adherence to associated arthritis, and in this case, timely targeted consultation of a specialist is necessary.

Synthesis and Analysis of Properties of an Immunogenic Fragment from NS4A Polypeptide of Hepatitis C Virus

Abstract— The work is aimed at the synthesis and analysis from NS4A of hepatitis C virus (HCV) antigen peptide fragment that contains a conserved B-cell and T-helper epitopes. The 24-mer peptide VIVGRIILSGRPAVIPDREVLYRK-NH2, which contains the main immunogenic site 24–46 of HCV NS4A antigen (corresponding to the 1681–1703 amino acid residues of the HCV polypeptide), subtype 1b, has been prepared via solid-phase synthesis according to the Fmoc-protocol. Particles with diameters of 73 ± 10 nm (30%) and 236 ± 5 nm (70%) have been detected in the water solution of the highly purified peptide (0.5 mg/mL) by dynamic light scattering. The polydispersity index of 0.377 ± 0.012 implies the existence of heterogeneity because of the aggregation of the peptide molecules. The ζ-potential of the peptide aggregates has been determined as 7.0 ± 0.5 mV by means of electrophoretic light scattering. These data confirm the possibility for the development of a nanoscale liposome form of the peptide preparation. Immunoreactivity of the synthesized highly purified peptide has been studied with the use of blood sera of patients with chronic hepatitis C. Antipeptide immunoglobulins G have been detected in 41.7% of serum samples. Thus, this peptide has been shown to reproduce at least one B-epitope, to which antibodies are raised during natural HCV infection. The synthesized 24-mer peptide is a promising candidate for further research and for use as a potential immunogen for the design of a nanoscale therapeutic immunogenic liposomal peptide composition with synthetic lipids as an adjuvant.

Phylogenetic and Molecular Analyses of More Prevalent HCV1b Subtype in the Calabria Region, Southern Italy

Hepatitis C virus subtype 1b (HCV1b) is still the most prevalent subtype worldwide, with massive expansion due to poor health care standards, such as blood transfusion and iatrogenic procedures. Despite safe and effective new direct antiviral agents (DAA), treatment success can depend on resistance-associated substitutions (RASs) carried in target genomic regions. Herein we investigated transmission clusters and RASs among isolates from HCV1b positive subjects in the Calabria Region. Forty-one NS5B and twenty-two NS5A sequences were obtained by Sanger sequencing. Phylogenetic analysis was performed using the maximum likelihood method and resistance substitutions were analyzed with the Geno2pheno tool. Phylogenetic analysis showed sixteen statistically supported clusters, with twelve containing Italian sequences mixed with foreign HCV1b isolates and four monophyletic clusters including only sequences from Calabria. Interestingly, HCV1b spread has been maintained by sporadic infections in geographically limited areas and by dental treatment or surgical intervention in the metropolitan area. The L159F NS5B RAS was found in 15 isolates and in particular 8/15 also showed the C316N substitution. The Y93H and L31M NS5A RASs were detected in three and one isolates, respectively. The A92T NS5A RAS was found in one isolate. Overall, frequencies of detected NS5B and NS5A RASs were 36.6% and 22.7%, respectively. For the eradication of infection, improved screening policies should be considered and the prevalence of natural RASs carried on viral strains.

Identification and genotyping of a new subtype of bovine viral diarrhea virus 1 isolated from cattle with diarrhea

In 2019, diarrhea cases occurred on cattle farms in Qionglai and Guang'an, Sichuan Province. Two out of 20 (10%) serum and nasal swab samples were positive when tested using a bovine viral diarrhea virus (BVDV) antigen-capture ELISA kit. Two non-cytopathic strains of BVDV were isolated and named QL1903 and GA190608, respectively. The nucleotide sequences of the genomes of the two isolates were 89.52% identical. Phylogenetic analysis based on the 5'-UTR sequence revealed that the BVDV isolate QL1903 belonged to BVDV subtype 1b, whereas isolate GA190608 clustered with strains HN1814, EN-19, and BJ09_26 in a separate branch, which has tentatively been classified as a new genetic subtype, "1v".

Evolutionary modelling of HCV subtypes provides rationale for their different disease outcomes

Hepatitis C virus (HCV) is a leading cause of liver-associated disease and liver cancer. Of the major HCV subtypes, patients infected with subtype 1b have been associated with having a higher risk of developing chronic infection, cirrhosis and hepatocellular carcinoma. However, underlying reasons for this increased disease severity remain unknown. Here, we provide an evolutionary rationale, based on a comparative study of fitness landscape and in-host evolutionary models of the envelope glycoprotein 2 (E2) of HCV subtypes 1a and 1b. Our analysis demonstrates that a higher chronicity rate of subtype 1b may be attributed to lower fitness constraints, enabling 1b viruses to more easily escape antibody responses. More generally, our results suggest that differences in evolutionary constraints between HCV subtypes may be an important factor in mediating distinct disease outcomes. Our analysis also identifies antibodies that appear to be escape-resistant against both subtypes 1a and 1b, providing directions for the design of HCV vaccines having cross-subtype protection.

THE IMPORTANCE OF ANTI-HCV SCREENING IN PATIENTS OF LARGE MULTIFIELD HOSPITALS

In the studied cohort, HCV subtypes 1b and 3a prevailed — 47,7% and 38,9%, respectively HCV genotype 2 was detected in 7,2% of cases and HCV subtype 1a — in 5,8% of cases. HCV genotype was not determined in 0,4% of patients. Difference between a frequency of detection of subtypes 1b and 3a of HCV in men and women were recorded. Subtype 3a HCV was determined in men in 44,8% of cases, and in women — in 31,7% of cases (p < 0.01). HCV subtype 1b in women was determined with a higher frequency (55,0%, р < 0, 01), than in men (41,8%).

Phylogenetic analysis confirms hepatitis C virus transmission among hemodialysis patients in Kosovo

Introduction: It has recently been demonstrated that there is a very high prevalence of hepatitis C virus (HCV) infection among hemodialysis patients in Kosovo with HCV subtype 1 being the most prevalent subtype. In this study, we further detail the molecular epidemiology of HCV outbreaks occurring in seven dialysis centers in Kosovo. Methodology: In total, 273 samples obtained from HCV RNA positive patients undergoing hemodialysis at one of the seven centers in Kosovo were selected for this study: 171 subtype 1a samples, 91 subtype 4d samples, and 11 subtype 1b samples. A partial HCV NS5B region was amplified and sequenced. Subtype-specific phylogenetic analyses were performed with the inclusion of control sequences and transmission clusters were identified. Results: NS5B sequences were successfully obtained in 257/273 (94.1%) of samples; 162 subtype 1a, 84 subtype 4d, and 11 subtype 1b sequences. Phylogenetic analyses showed a high degree of phylogenetic clustering of HCV sequences subtyped 1a (99.4%), 1b (63.6%), and 4d (76.2%). Distinct phylogenetic clusters of sequences obtained from hemodialysis patients were observed for all three subtypes studied. In addition, several smaller clusters within the large clusters were identified, mainly from a single dialysis center. Conclusions: Phylogenetic analyses confirmed nosocomial transmission during dialysis as a major factor in the spread of HCV at the seven dialysis centers in Kosovo.