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Novel approach for quantification of hepatitis C virus in liver cirrhosis using real-time reverse transcriptase PCR
Kewal Krishan Maudar1, Puneet Gandhi, Pradyumna Kumar Mishra
1Department of Surgical Gastroenterology, Bhopal Memorial Hospital & Research Center, Karond, Raisen Bypass Road, Bhopal, 462 038, Madhya Pradesh, India. maudarji@yahoo.com
Insights
Hepatitis C virus (HCV) RNA was detected in liver tissue of three patients with serosilent HCV infection. This modified Light Cycler 2.0 method reliably detects and localizes HCV RNA in liver biopsies.
Area of Science:
- Hepatology
- Virology
- Molecular Diagnostics
Background:
- Hepatitis C virus (HCV) infection affects 3% of the global population, leading to liver fibrosis, cirrhosis, and cancer.
- Current HCV diagnosis relies on detecting anti-HCV antibodies in serum.
- Detecting HCV RNA in liver tissue is crucial for diagnosing HCV, especially when serum markers are negative.
Purpose of the Study:
- To evaluate a modified Light Cycler 2.0 technique for quantitative detection and localization of HCV RNA in liver tissue.
- To assess the reliability of this method in diagnosing "serosilent" HCV infections.
Main Methods:
- Ten liver biopsy samples from patients with cryptogenic liver cirrhosis, negative for anti-HCV and serum HCV RNA, were analyzed.
- HCV RNA detection in liver tissue was performed using a modified protocol with Light Cycler 2.0, achieving a minimum detection limit of 10 copies/ml.
Main Results:
- HCV RNA was quantitatively detected in three out of ten liver biopsy samples.
- The modified method demonstrated the presence of viral nucleic acid in liver tissues.
Conclusions:
- The modified Light Cycler 2.0 technique provides a reliable method for quantitative detection and localization of HCV RNA in liver tissue.
- This approach is effective for diagnosing cases of "serosilent" HCV infection where traditional serological markers are absent.
Background:
Hepatitis C virus (HCV) infects nearly 3% of the population worldwide and is a major cause of acute and chronic infections leading to fibrosis, cirrhosis, and hepatocellular carcinoma. Current laboratory diagnosis of HCV is based on specific antibody detection (anti-hepatitis C virus (anti-HCV)) in serum. As HCV replicates in the liver cells, detection and localization of HCV RNA in liver tissue are vital for diagnosis.
Methods:
Ten biopsy samples diagnosed for cryptogenic liver cirrhosis, negative for the presence of anti-HCV and serum HCV RNA, were studied for analyzing presence of viral nucleic acid in liver tissues. Qualitative screening for HCV was done through ELISA while the nucleic acid analysis was performed through COBAS Amplicor. Detection of HCV RNA in liver tissue biopsies was performed following standard protocol of HCV detection kit (Shenzhen PG Biotech) with modifications using Light Cycler 2.0 (minimum detection limit 10 copies/ml).
Result:
Quantitative detection in liver biopsies following the modified method showed the presence of HCV RNA in three samples out of the ten studied.
Conclusion:
The results indicate that using Light Cycler 2.0, following the modified technique described, constitutes a reliable method of quantitative detection and localization of HCV in tissue in "serosilent" HCV infection.
