*Dr. Mushtak T. S Al-Ouqaili **Dr. Yasin Hamad Majeed Based on Molecular level, A Study of Hepatitis C in AL-Anbar Governorate, West of Iraq By:- *Dr. Mushtak T. S Al-Ouqaili **Dr. Yasin Hamad Majeed *Assistant Professor, Dean of College of Pharmacy, University of Al-Anbar **Lecture, Dep. Of Intern. Med., College of Medicine, University of Al-Anbar

بِسمِ اللهِ الرَّحمن الرَّحِيمِ {فَأَمَّا الزَّبَدُ فَيَذْهَبُ جُفَاءً وَأَمَّا مَا يَنفَعُ النَّاسَ فَيَمْكُثُ فِي الأَرْضِ كَذَلِكَ يَضْرِبُ اللَّهُ الأَمْثَال} صَدَقَ اللهُ العَظيمْ

Introduction

Introduction Hepatitis C virus (HCV) infection is one of the main causes of chronic liver disease worldwide. It is estimated that approximately 130–210 million individuals i.e. 3% of the world population, are chronically infected with HCV.

Hepatitis C Virus

HCV Structure RNA virus Virion: Single-stranded Enveloped Positive-sense 9400 nucleotide Virion: Enveloped Icosahedral capsid 40-60 nm in diameter

HCV Genome 1 hypervariable region: E1, E2 envelope proteins Allows for evasion of host immunologic response 9400 nucleotide, single ORF Codes for a viral polyprotein about 3000 a.a. in length 3 highly conserved areas: 5’ UTR: initiating translation Core: codes for capsid protein monomer 3’ UTR: essential for RNA synthesis & packaging

Pathology of HCV Acute Hepatitis C: Generally benign: No jaundice (80%) Usually asymptomatic Can be severe, but liver failure rare Only real threat of acute Hepatitis C is its ability to reach chronic stages undetected and untreated.

Pathology of HCV Chronic Hepatitis C: 70% of patients become chronic Possible results: Cirrhosis End-stage liver disease Hepatocellular carcinoma

Aims of this study

Aims of this study This study has been undertaken for the following purposes:- 1:- Preliminary screening for hepatitis C virus infection depending on immunochromatographic essay and enzyme linked immunosorbent essay. 2:- Detection of HCV-RNA copies through determination of viral load by real time polymerase chain reaction (RT-PCR)

Aims of this study 3:- Genotyping of HCV-RNA using Smart Cycler Genotyper. 4:- Follow up of Patients infected with hepatitis C virus before and after treatment using quantitative real time PCR.

Patients & methods

Method Plan Laboratory Part Serological Part Molecular Part

Methods A total of 105 patients with suspicion of hepatitis C infection were studied during the period from January, 2012 and till now. They were well diagnosed by experient clinician during admitting GIT Centre in Ramadi Teaching Hospital and Private Clinics.

The clinical samples were submitted for immunochromatographic essay and enzyme linked immunosorbent (ELISA) essay as preliminary screening tests.

Confirmatory test include reverse transcriptase real time polymerase chain reaction (RT-PCR) were used to detect viral load for HCV-RNA copies considering the lower sensitivity threshold was 200 copy/ml in addition to follow up the quantity of nucleic acid copies after period of therapeutic regimen using the same instrument.

Also, advanced molecular technique include genotyping to detect HCV-RNA genotypes into (1, 2, 3, 4, 5, and 6) was achieved.

Results & Discussion

Results and Discussion The study result revealed that all study patients, 105 (100%) were positive by dipstick assay which depends on its mechanism on immunochromatographic essay. Of these, 91 (86.7%) patients were appeared to be positive for anti-hepatitis C virus antibodies by ELISA. the study patients

Results and Discussion Out of these patients, 73 (80.2%) had observed number of HCV-RNA by RT-PCR with mean 5.34E+06 ± 5.34E+06 IU/ml (1.81E+07 ± 3.87E+07 copy/ml) while the remaining 18 (19.8%) showed undetectable HCV-RNA copies (Less than lower sensitivity threshold, 200 copy/ml).

Statistical parameters   Mean 5.34E+06 1.81E+07 Median 1.22E+06 4.88E+06 Std. Deviation 1.22E+07 3.87E+07 Minimum 1.70E+02 6.80E+02 Maximum 7.49E+07 2.18E+08

Results and Discussion In genotyping study, positive HCV-RT-PCR patients were yielded as follow: 56 (76.7%) with genotype 4, 14 (19.2%) genotype 1 while 3 (4.2%) were positive for genotype 3. On the other hand, in term of quantitative RT-PCR, out of study patients, 19 (20.8%) were submitted to follow up before and after treatment with ribavirin and interferon. They showed sharp decrease in HCV-RNA copies count.

In Iraq, the prevalence of HCV ranged from 3. 21 to 7 In Iraq, the prevalence of HCV ranged from 3.21 to 7.1% among the pregnant women and general population of four HCV genotypes, 1a, 1b, and 3a and 4. A study carried out on Iraqi refuges showed further discrepancy in this regard; The researcher showed that by using HCV-RNA analysis found that the prevalence among these refuges was 0.1%, and HCV genotype 4c, d was the commonest.

HCV is less frequent among Lebanese; it ranged from 0. 16 to 1 HCV is less frequent among Lebanese; it ranged from 0.16 to 1.22% among healthy volunteers, blood donors, and health care workers, though it was 27% among haemodialysis patients. Genotype 4 was the commonest isolated genotype among the various groups studied followed by genotypes 1a and 1b. The above results were in consistent with our study results.

Conclusions & Recommendations

Conclusions ELISA was reliable confirmatory test for detection of anti-hepatitis C virus antibodies. On molecular level, RT-PCR was a cut-off diagnostic tool for confirmation of HCV infection. 3-It is so beneficial in the follow up of patients to monitor therapeutic response depending on the amount of HCV-RNA copies. 4-HCV-genotype 4 was the most common among study patients followed by genotype 1 and 3 respectively.

Aknowledgment Special thanks and deep gratitude to my colleague the experient gastroenterologist Dr. Yasin Hamad Majeed, Lecturer in Department of Internal Medicine, College of Medicine, University of Anbar for his scientific efforts in providing the work clinical specimens and and scientific contribution in all study details

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