Levels of hepatitis B virus replicative intermediate in serum samples of chronic hepatitis B patients

Abstract

Hepatitis B virus (HBV) cccDNA levels is an absolute marker of HBV replication in the liver of HBV infected patients. This study aimed to quantify the HBV cccDNA levels in sera and liver tissue samples of treatment naïve patients with chronic hepatitis B. Eighty one chronic hepatitis B (CHB) treatment naïve patients were enrolled from January 2009 to June 2011. Total HBV DNA and HBV cccDNA levels were quantified using sensitive real time PCR assay. The mean age of recruited patients was 34 ± 11.5 years. Fifty four (66.7%) patients were HBeAg negative. Liver tissue samples were available from 2 HBeAg positive and 21 HBeAg negative CHB patients. The amount of total intrahepatic HBV DNA ranged from 0.09 to 1508.92 copies/cell. The median intrahepatic HBV cccDNA was 0.31 and 0.20 copies/cell in HBeAg positive and HBeAg negative cases, respectively. Serum HBV cccDNA was detectable in 85.2 % HBeAg positive and 48.1% HBeAg negative CHB patients. Median serum HBV cccDNA was 46,000 and 26,350 copies/mL in HBeAg positive and HBeAg negative subjects, respectively. There was a significant positive correlation between the levels of intrahepatic total HBV DNA and intrahepatic HBV cccDNA (r = 0.533, p = 0.009). A positive correlation was also seen between serum HBV cccDNA levels and serum HBV DNA levels (r = 0.871, p < 0.001). It was concluded that serum HBV cccDNA could be detectable in higher proportion of HBeAg positive patients compared to HBeAg negative patients. Moreover, the median level of serum HBV cccDNA was significantly higher in HBeAg positive patients in contrast to HBeAg negative subjects.

Detection of Hepatitis B Virus Covalently Closed Circular DNA in the Plasma of Iranian HBeAg-Negative Patients With Chronic Hepatitis B

“…It has been reported that serum cccDNA of HBV was detectable in 85.2% HBeAg positive and 48.1% negative chronic hepatitis B participants ( 12 ), whereas the HBV cccDNA was detected in plasma specimens in 19 (17.9%) out of the total 106 patients in the present study. Therefore, it seems that more research should be done on this area.…”

“…The main disadvantage of detection of the HBV cccDNA before treatment response is the requirement for liver biopsies ( 16 ). However, it has been reported that the HBV cccDNA can also be detected in the plasma of HBV-infected patients ( 12 , 15 ).…”

“…Therefore, the detection of cccDNA should be performed on other samples, when a liver biopsy specimen is not available. Recent studies have indicated that the HBV cccDNA level is a marker of HBV replication in the hepatocytes of HBV-infected patients ( 12 ).…”

Oral administration of encapsulated bovine lactoferrin protein nanocapsules against intracellular parasite Toxoplasma gondii

“…The number of copies per unit volume (mL) was calculated using the formula described elsewhere. 25 Serial dilutions of these stock solutions served as quantification standards to plot the standard curves. Following formula was used to calculate the copy number.…”

Detection of HBV Covalently Closed Circular DNA

“…Some reports have shown that cccDNA is present in patients’ sera and that it is a marker of off-treatment virological relapse [11,26,32,34,114], whereas others have refuted the existence of cccDNA in serum and have used DNA extracted from CHB patients’ sera as a negative control in the quantification of intracellular cccDNA [39,42,63]. Do some non-liver cells, such as PBMCs, support HBV infection and formation of cccDNA?…”