Innovative technologies in the diagnosis of chronic hepatitis B.


Chronic hepatitis B is one of the main etiological causes of liver failure, cirrhosis and hepatocellular carcinoma worldwide. This condition cannot be completely cured by currently available medications due to the constant existence of the ring covalently closed DNA (ccDNA) of the hepatitis B virus (HBV) in infected hepatocytes, which is the matrix for transcription of HBV RNA. Since the quantitative assessment of CDDNA itself requires an invasive procedure (liver biopsy), serum biomarkers reflecting intrahepatic activity of CDDNA are needed. Recently, more and more studies show that circulating HBV RNA can serve as a serum biomarker of infection, treatment and prognosis of HBV. Serum RNA is transcribed directly from cDNA, so serum RNA levels may indicate the presence of transcriptional activity of the virus.

HBV RNA levels correlate with HBV DNA levels and are associated with serum HBsAg and ALT levels. HBV RNA levels vary depending on the infection phase — they are highest in the immunotolerant phase and lowest in patients with inactive HBV infection. In patients receiving treatment with nucliazide analogues (the main group of drugs used to treat chronic viral hepatitis B), HBV RNA levels are higher than HBV DNA levels. In general, HBV RNAs in serum indicate transcriptional activity of CDDNA. This biomarker (in combination with HBV DNA and HBsAg levels) can be used to monitor the response to treatment in patients receiving therapy with nucliazide analogues.

It can also be used to identify patients who can safely stop taking nucleazide analogues and/or achieve the disappearance of HBsAg (functional cure).Serial evaluation of intrahepatic HBV cDNA is not possible, therefore, measurement of HBV RNA together with analysis of viral activity can be used to determine the activity of HCDNA, especially when developing treatment methods aimed at elimination and/or functional cure.

ROSSA has developed a set of reagents “ROSSAmed HBV-RNA” for the detection of Hepatitis B virus RNA by polymerase chain reaction in real time. The kit is suitable for tablet-type devices: “DTprime”, “Dtlight”, “DT-96” (NPO DNA Technology LLC), or CFX96 (Bio-Rad), qTOWER (Analytik Jena), as well as for a rotary-type detecting amplifier, for example, Rotor-Gene Q (QIAGEN).

The ROSSAmed HBV- RNA kit was tested in the reference laboratory of the M.Z. RUz Virology Research Institute, and also received a registration certificate of the “State Center for Expertise and Standardization of Medicines, Medical Devices and Medical Equipment” of the Republic of Uzbekistan. The kit set received positive conclusions and was recommended for use in laboratory practice. There are no analogues of the ROSSAmed HBV- RNA kit in Uzbekistan at the moment.

The recommended system for isolating RNA/DNA from biological material is “MagSorb Plasma” (manufactured by OOO “ROSSA”).