Hepatitis C virus genotype frequency in Isfahan province of Iran: a descriptive cross-sectional study

<p>Abstract</p> <p>Background</p> <p>Hepatitis C is an infectious disease affecting the liver, caused by the hepatitis C virus (HCV). The hepatitis C virus is a small, enveloped, single-stranded, positive sense RNA virus with a large genetic heterogeneity. Isolates have been classified into at least eleven major genotypes, based on a nucleotide sequence divergence of 30-35%. Genotypes 1, 2 and 3 circulate around the world, while other genotypes are mainly restricted to determined geographical areas. Genotype determination of HCV is clinically valuable as it provides important information which can be used to determine the type and duration of therapy and to predict the outcome of the disease.</p> <p>Results</p> <p>Plasma samples were collected from ninety seven HCV RNA positive patients admitted to two large medical laboratory centers in Isfahan province (Iran) from the years 2007 to 2009. Samples from patients were subjected to HCV genotype determination using a PCR based genotyping kit. The frequency of HCV genotypes was determined as follows: genotype 3a (61.2%), genotype 1a (29.5%), genotype 1b (5.1%), genotype 2 (2%) and mixed genotypes of 1a+3a (2%).</p> <p>Conclusion</p> <p>Genotype 3a is the most frequent followed by the genotype 1a, genotype 1b and genotype 2 in Isfahan province, Iran.</p

Allosteric Modulation of the HIV-1 gp120-gp41 Association Site by Adjacent gp120 Variable Region 1 (V1) N-Glycans Linked to Neutralization Sensitivity

The HIV-1 gp120-gp41 complex, which mediates viral fusion and cellular entry, undergoes rapid evolution within its external glycan shield to enable escape from neutralizing antibody (NAb). Understanding how conserved protein determinants retain functionality in the context of such evolution is important for their evaluation and exploitation as potential drug and/ or vaccine targets. In this study, we examined how the conserved gp120-gp41 association site, formed by the N- and Cterminal segments of gp120 and the disulfide-bonded region (DSR) of gp41, adapts to glycan changes that are linked to neutralization sensitivity. To this end, a DSR mutant virus (K601D) with defective gp120-association was sequentially passaged in peripheral blood mononuclear cells to select suppressor mutations. We reasoned that the locations of suppressors point to structural elements that are functionally linked to the gp120-gp41 association site. In culture 1, gp120 association and viral replication was restored by loss of the conserved glycan at Asn136 in V1 (T138N mutation) inconjunction with the L494I substitution in C5 within the association site. In culture 2, replication was restored with deletion of the N139INN sequence, which ablates the overlapping Asn141-Asn142-Ser-Ser potential N-linked glycosylation sequons inV1, in conjunction with D601N in the DSR. The 136 and 142 glycan mutations appeared to exert their suppressive effects by altering the dependence of gp120-gp41 interactions on the DSR residues, Leu593, Trp596 and Lys601. The 136 and/or 142glycan mutations increased the sensitivity of HIV-1 pseudovirions to the glycan-dependent NAbs 2G12 and PG16, and also pooled IgG obtained from HIV-1-infected individuals. Thus adjacent V1 glycans allosterically modulate the distal gp120-gp41 association site. We propose that this represents a mechanism for functional adaptation of the gp120-gp41 association site to an evolving glycan shield in a setting of NAb selection

Profit enhancing competitive pressure in vertically related industries

Coevolution of viruses and their hosts represents a dynamic molecular battle between the immune system and viral factors that mediate immune evasion. After the abandonment of smallpox vaccination, cowpox virus infections are an emerging zoonotic health threat, especially for immunocompromised patients. Here we delineate the mechanistic basis of how cowpox viral CPXV012 interferes with MHC class I antigen processing. This type II membrane protein inhibits the coreTAP complex at the step after peptide binding and peptide-induced conformational change, in blocking ATP binding and hydrolysis. Distinct from other immune evasion mechanisms, TAP inhibition is mediated by a short ER-lumenal fragment of CPXV012, which results from a frameshift in the cowpox virus genome. Tethered to the ER membrane, this fragment mimics a high ER-lumenal peptide concentration, thus provoking a trans-inhibition of antigen translocation as supply for MHC I loading. These findings illuminate the evolution of viral immune modulators and the basis of a fine-balanced regulation of antigen processing

S-gene sequences and genotype-related restriction sites in hepatitis B virus carriers in Turkey

Background: Although hepatitis B virus (HBV) infections are a major health problem in Turkey, there is little information on the genotype distribution of the virus. In this study, HBV genotypes were determined by DNA sequencing and restriction enzyme analysis of the S gene

Sequence analysis of the 5' non-coding region of Turkish HCV isolates: Implications for PCR diagnosis

Hepatitis C virus (HCV) is a positive-strand RNA virus related to pestiviruses and flaviviruses. The 5' noncoding region (NCR) of the virus genome consists of 324-341 nucleotides and is generally highly conserved among different HCV isolates which has made this region the choice for primer selection in amplification of HCV sequences by polymerase chain reaction (PCR). In this study, we report the partial nucleotide sequences of the 5'-NCR from type la (n = 4), type 1b (n = 6) and type 4 (n = 1) Turkish HCV isolates. Sequence information was obtained by direct sequencing of RT-PCR product using biotinylated primers and single strands were sequenced using T7 DNA polymerase after binding to streptavidin coated magnetic beads. In comparison to prototype type la consensus sequence, all type 1b sequences had A-G substitution at position - 99. Nucleotid changes from the prototype 1a sequence were found in 12 of the 174 nucleotide positions. The most variable domain spans 51 nucleotides (positions - 167 to - 117) where nine polymorphic sites were identified. Although the nucleotide sequence of the 5'-noncoding region is highly conserved there are type-specific polymorphic sites within this region that has to be taken into consideration in the design of oligonucleotide primers for reliable amplification of sequences from different HCV genotypes

CONCENTRATION AND AVIDITY OF ANTITETANUS ANTIBODIES IN MOTHER-INFANT PAIRS - RELATION TO IMMUNIZATION TIME

The concentration and avidity of anti-tetanus antibodies in two groups of mother-infant pairs were compared. Mothers immunized during pregnancy and their newborns (group A) had significantly higher antibody concentrations than mothers immunized at least a year before their last pregnancy and their newborns (group B) as measured by an indirect enzyme immunoassay (EIA) procedure. Antibody avidity of samples was measured by an inhibition EIA technique and urea denaturation test. Although antibody avidity was higher in group B, the differences were not significant. These findings may represent a secondary antibody response to a protein antigen, when considering that all mothers in both groups had received a primary tetanus vaccination during childhood. In mothers with a history of primary tetanus immunization, a single booster dose of tetanus toroid during pregnancy is enough to induce protective levels of antibodies with reasonably high avidity in both mother and newborn

Molecular evidence of nosocomial transmission of hepatitis C virus in a haemodialysis unit

The molecular epidemiology of type 2a hepatitis C virus (HCV) infections in patients undergoing haemodialysis in the same unit in a Turkish hospital was investigated. Of nine HCV-infected patients four were infected with type 2a, four with type Ib and one with type la viruses. Since type 2 HCV infections in the Turkish population are rare, the possibility of nosocomial infection was investigated by means of phylogenetic analysis of viral sequences amplified by the polymerase chain reaction in the NS5b region. One of the samples failed to show amplification and therefore could not be sequenced. The sequences of the remaining three virus samples were grouped closely in a cluster within the type 2a group. The results thus showed that three patients were infected with the same HCV type 2a strain. Seroconversion and clinical data suggested that these patients may have been infected on different occasions, there being possibly more than one mode of transmission. Breaches in infection control procedures and lack of environmental decontamination between two haemodialysis sessions were probably the causes of HCV infections in these patients

Application of hepatitis serology testing algorithms to assess inappropriate laboratory utilization

Rationale, aims and objectives Many studies pointed out inappropriate utilization of laboratory caused by excessive amounts of tests ordered by doctors. To prevent and to eliminate the ordering of unhelpful tests, introducing diagnostic algorithms, which are also a suitable practice for application to hepatitis serology, have been suggested. This study aimed to determine inappropriate test ordering rates with respect to the commonly approved algorithms for serological diagnosis of viral hepatitis. Methods To assess the number of inappropriate test orders, laboratory records of samples sent for hepatitis A, B, and D serology were reviewed and evaluated retrospectively with respect to algorithms for serological diagnosis of viral hepatitis. Orders including serological marker groups with inadequate clinical information to determine whether or not the order was inappropriate were excluded from the analysis. Results Application of diagnostic algorithms showed that 50% of anti-HAV IgM and anti-HAV total; 12.7% of anti-HBs, 12.7% of anti-HBc total, 78.5% of anti-HBc IgM, 87.3% of HBe Ag, 78.8% of anti-HBe, 58.7% of anti-HD total orders were made inappropriately. Conclusions Our study provides information for inappropriate laboratory utilization for hepatitis serology testing and we suggest to use diagnostic algorithms applied by the serology laboratory to decrease the rate of unhelpful test orders