We are currently trying to get the complete genome of the hepatitis B virus using the Gunther method for PCR. Usually, this methodology is useful for a HBV viral load of more than 10000. However, in occult hepatitis B patients, the viral load is lower than 10000. we have been trying to use the Gunther method with a different thermal profile. Until now, we were not able to get the complete genome in PCR.
You may follow the protocol described in the following publication and amplify Occult HBV genome:
Chaudhuri V, et al. (2004). Occult hepatitis B virus infection in chronic liver disease: full-length genome and analysis of mutant surface promoter.
Gastroenterology 127: 1356-1371.
our own primers which had been designed in Dr Carman's lab in Scotland long time ago (90s) are quiet efficient (Jazayeri et al, 2004-Journal of Viral Hepatitis). after increasing the amount of initial temple and an Mgcl2 gredient in multiple runs, we could be able to get about 90% of our occult HBV DNA. only 6 samples left.
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