الفهرس 
Review of LiteratureOnly 14 pages are availabe for public view
Abstract
Hepatitis C virus (HCV) infection has been recognized by WHO as a global health burden and a public health threat since 2016. One hundred and thirty to one hundred and fifty million hepatitis C virus-infected patients occur all over the world and up to 80% become chronic cases and many of them suffer from liver cirrhosis and hepatocellular carcinoma (HCC). About 700,000 deaths occur yearly due to hepatitis C-related liver diseases worldwide.
Although hepatitis C virus is mainly hepatotropic, it can infect peripheral blood mononuclear cells (PBMCs) and remain there as a reservoir. Persistence of replicating virus in PBMCs may cause reactivation of infection later on even after viral clearance from the serum. A varying number of HCV infected patients have experienced relapses after achieving SVR post-treatment with DAAs.
This study aimed to detect HCV RNA by PCR in PBMCs in patients with chronic hepatitis C after antiviral therapy and compare that with viral detection in the serum to determine if the presence of viral RNA in PBMCs after apparent clearance from serum samples can be used as a predictor of relapse after treatment.
In this study, fifty-two chronic infected HCV patients (22 males and 30 females) with ages ranging from 18 to 75 years old, were selected from Ain Shams University Hospitals in the period from December 2016 to March 2017. All the enrolled patients were positive for HCV RNA by real-time PCR before treatment. Patients whose INR ≥ 1.7, serum bilirubin > 3mg/dl, serum albumin < 2.8 g/dl, platelets count < 50000 /mm3, with hepatocellular carcinoma, with extra-hepatic malignancy and pregnant were excluded.
According to Egyptian National Committee for Control of Viral Hepatitis, November 2015, patients were classified into two groups:
’’Easy to treat’’ group (35 patients): treatment-naive patients whose total serum bilirubin ≤1.2mg/dl, serum albumin ≥ 3.5g/dl, INR ≤ 1.2 and platelet count > 150000/mm3. They were treated with sofosbuvir (400mg/day) and daclatasvir (60mg/day).
’’Difficult to treat’’ group (17 patients): Peg-IFN (Pegylated Interferon) treatment-experienced patients whose total serum bilirubin ≥ 1.2mg/dl, serum albumin ≤ 3.5g/dl, INR ≥ 1.2 and platelet count ˂ 150000/mm3. They were treated with sofosbuvir (400mg/day), daclatasvir (60mg/day) and weight-based ribavirin.
All patients underwent the following:
a. Baseline screening: Full clinical history including history of schistosomiasis infection, blood transfusion, surgery, being a member of healthcare workers and a history suggestive of liver cell failure. Laboratory tests included complete blood count, liver and kidney function tests, Alpha Feto protein levels, HCV Ab, HBVs Ag and real time quantitative PCR for serum HCV RNA levels.
b. Collection of blood samples at three interval times; at the end of treatment (EOT), 12 weeks, and 52 weeks later.
c. End of treatment (EOT) assessment: Clinical and laboratory examinations represented monthly follow up. In additional to HCV RNA detection in both serum and PBMCs, through HCV RNA extraction first, then amplification by nested RT-PCR in PBMCs and quantification of HCV RNA in sera by Real- time PCR.
d. Twelve weeks and 52 weeks after EOT follow up: Clinical and laboratory examinations were similar to the monthly follow up. Additionally, serum HCV RNA was determined by PCR method.
The results demonstrated that all fifty-two patients (100%) achieved ETR and SVR. Forty-one patients (78.8%) showed undetectable HCV RNA in PBMCs, while 11 patients (21.2%) showed positive HCV RNA in PBMCs at EOT. Six of these patients (11.54%) relapsed (showing detectable HCV RNA in serum) 52 weeks after the EOT.
A statistically significant difference was found between relapsers and non-relapsers as regards HCV RNA-positivity/negativity in their PBMCs. All relapsers had positive HCV RNA in their PBMCs (P<0.001). There is no statistically significant difference between relapsers and non-relapsers as regards treatment regimen (P=1) gender of patients (P=0.226), and any of the studied baseline characteristics.
The performance of HCV RT PCR in PBMCs in comparison to HCV RT PCR in serum as gold standard test in prediction of HCV relapsers; its sensitivity is 100%, specificity is 89%, PPV is 54.6% and NPV is 100% with accuracy of the test is nearly 90%.
These findings indicate that the absence of HCV in the serum of chronic HCV infected patients does not mean that there is no circulating virus. HCV RNA in mononuclear cells may be an indicator of persisting infection and can therefore be a predictor of relapse. Considering the significance of viral persistence and subsequent relapse following treatment in the control of HCV infection, this study warrants attention to viral detection in PBMCs as a mechanism of patient follow-up.