Hepatitis Monthly
Volume:20 Issue: 9, Sep 2020

Although the liver is the main site for the Hepatitis C Virus (HCV) replication, there is still an essential debate about extrahepatic HCV reservoirs.

It has been proposed that Peripheral Blood Mononuclear Cells (PBMCs) could be the possible virus replication sites. Therefore, PBMCs may be candidates for recurrent HCV infection after achieving Sustained Virologic Response (SVR). In this study, we designed a lymphocyte culture to explore more about virus replication in PBMCs collected from patients with chronic hepatitis C.

Plasma and PBMC samples were collected from 16 randomly selected seropositive patients for the anti-HCV antibody. Four out of 16 (25%) patients received combination therapy with alpha interferon and ribavirin. PBMCs were isolated from whole blood. Between 106 -107 cells were cultured with optimized concentrations of IL-2 (10 mg/ml) and phytohemagglutinin A (5 mg/ml). Total RNA was extracted from the first collected sera and harvested lymphocytes. Constructed plasmids containing the NCR coding region were used to plot the standard curve for the relative quantification of SYBR green real-time PCR. The sensitivity and specificity of the detection were established by using plasmids containing cDNA.

With this plasmid containing the NCR coding region, the Limit of Detection (LOD) of in-house-developed real-time RT-PCR sensitivity was 2×101 copies. Using primers for the NCR region, 10 out of 16 (62.5 %) PBMCs were positive for negative-strand HCV RNA. Among the four samples collected from patients with SVR, negative-strand HCV RNA was found in two patient samples.

Our results indicated that cultured lymphoid cells from patients with chronic hepatitis, even with SVR, in the presence of IL-2 and PHA, markedly enhanced the detection of HCV RNA replica-tive strands. Therefore, PBMCs may be reservoirs for recurrent hepatitis infection after SVR and antiviral treatment. However, more clinical samples and control groups (lymphocyte culture without mitogen) should be examined to support the data presented in this study

With the identification of blood donors with a subsequent positive result in HCV screening and the possibility of HCV transmission via their previous negative donations, the role of look-back investigation has been crucial in enhancing blood safety. A retrospective study was conducted to identify the fate of blood components from donors subsequently found to be confirmed HCV positive cases with previous negative donations.

This look-back study aimed to determine the transfusion-induced hepatitis C virus infection through previous HCVnegative donations of subsequent HCV positive blood donors in Iran.

In this study, all serologically confirmed HCV positive blood donors across the country from December 2015 to June 2017 were included. A look-back process was conducted by tracing back previous HCV negative donations of subsequent HCV-positive blood donors to specific recipients, according to the IBTO instructions. The HCV RNA testing was carried out using an in-house onestep TaqMan real-time RT-PCR assay.

During the study period, 280 serologically confirmed HCV-positive blood donors were included, with 267 as first-time, nine as repeated, and three as regular donors. Of the participants, the first-time donors were excluded, the repeated donors were not eligible, and only the regular donors were selected for the look-back study. The HCV RNA was detected in one regular donor, but HCV infection was not detected in his previous donation recipient. In addition, more than two HCV risk factors were reported by each regular blood donor.

According to the results, evidence of HCV transmission via previous blood transfusions and the risk of HCV infection was not observed among recipients. Due to the reporting of some HCV risk factors by the subjects, paying proper attention to the donor selection process is highly recommended.

The serological measurement of the anti-hepatitis C virus antibody is a widely used tool in the first-line diagnosis of HCV infection. Therefore, increasing the testing criteria of these tests is of crucial importance for screening HCV infection.

The current study aimed to optimize a novel enzyme-linked immuno assay model to detect E2 antigen with or without sample pretreatment in combination with antibodies against core, NS3, NS4, and NS5 antigens of the hepatitis C virus and to compare the performances of these assays with indirect antigen (Ag), biotin/HRP labeled Antigen Sandwich andmethods of enzymelinked immunosorbent assay (ELISA) for their ability to detect HCV.

A total of 107 positive and 415 negative controls from volunteer whole blood donors in Blood TransfusionOrganization and 204 blood samples from patients under hemodialysis treatment in Tehran and Bandar Abbas hemodialysis centers are investigated. Six different methods of ELISA test were used to detect anti-HCV antibodies and/or HCV antigens in serum samples.

Regarding sensitivity, specificity, and accuracy, E2 Antigen detection alone or combined with antibody detection have the highest accuracy value (99% and 98%, respectively) compared to othermethods for antibodies detection. The results of the combined Ag/Ab ELISA test were closer to the results of real-time PCR.

This new approach to the detection of antigen and antigen/antibody has better performance criteria concerning the serologic detection of HCV, especially in HD patients who might experience a longer window period.

Hepatitis C is a public health threat, affecting approximately 1.9% of the Malaysian population.

This study demonstrates how a series of initiatives taken by the Ministry of Health (MOH) of Malaysia have impacted the treatment coverage and drug expenditure for hepatitis C patients since 2013, the year in which the first direct-acting antiviral (DAA) was introduced in public health institutions.

The data were obtained from all the 144 hospitals and 33 primary healthcare centers throughout the country, which were identified to have offered the pharmacological treatment to hepatitis C patients over the last seven years.

The total number of hepatitis C patients treated each year was shown to increase by more than 10 times, reaching 3,116 in 2019. However, the drug expenditure for hepatitis C relative to the overall health expenditure did not significantly increase over time (P = 0.094). The use of DAAs was once limited by its exorbitant cost. A remarkable elevation in the number of patients receiving the treatment only took place as of 2016, particularly following the engagement of the MOH in endeavors driven by non-profit organizations to enhance the accessibility of DAAs and the issuance of a compulsory license to sofosbuvir.

Timely decisions of the MOH and the judicious use of policy tools were shown to have transformed the landscape of hepatitis C management in Malaysia without considerably raising the budgetary pressure. Yet, continuous efforts to massively upscale the screening and treatment of the disease are warranted going forward.