Molecular diagnosis of viral hepatitis☆
Section snippets
Detection and quantification of viral genomes
Viral genomes are generally present in relatively small amounts in body fluids of infected patients, hindering their detection by simple molecular hybridization–based techniques. Thus, their detection and quantification require a preliminary “amplification” step. This can be achieved by using 2 categories of molecular biology–based techniques, namely target amplification and signal amplification.
HCV RNA detection and quantification
Table 1 lists the commercial assays that can currently be used to detect or quantify HCV RNA.Approval of each test for diagnostic purposes or research use only varies from one country to another at the time of writing.
Qualitative, nonquantitative assays can detect the presence of HCV RNA, but they cannot measure HCV viral load. They are based on target amplification, i.e., PCR or TMA. Table 1 shows the lower detection cutoffs of the available tests, as stated by the manufacturers. Qualitative
HBV DNA detection and quantification
Table 3 shows the commercial assays that can currently be used to detect and quantify HBV DNA.For each of them, current approval for diagnostic purposes or research use only varies from one country to another.
Table 3 shows the dynamic ranges of quantification stated by the manufacturers. The dynamic range of quantification can be extended to higher values by diluting and re-testing high-viral-load samples, but the dilution step may affect accuracy. These assays have been shown to be specific
Conclusion
In the past decade, the introduction and constant improvement of molecular biology–based techniques have provided invaluable tools for the management of chronic viral hepatitis. They can now be used to test blood donations, diagnose active infection, help to establish the prognosis, guide treatment decisions, and assess the virological response to therapy. Further work is required to fully standardize assays and quantification units, improve automation, and better define clinically relevant
Acknowledgements
The author is grateful to Magali Bouvier-Alias, Bertrand Pellegrin, and Susan L. Stramer for helpful discussion.
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Address requests for reprints to: Jean-Michel Pawlotsky, M.D., Ph.D., Department of Virology, Hôpital Henri Mondor, 51 avenue du Maréchal de Lattre de Tassigny, 94010 Créteil, France. e-mail: [email protected]; fax: 33 (1) 4981 2839.