Detection Of HBVDNA In PBMC With Occult HBV Infection And Analysis Of HBV S Gene Mutation

Background: HBsAg-positive HBV infection is an important evidence. However,there are a large number of clinical HBsAg-negative hepatitis. Studies have confirmedthat serum HBsAg negative in some individuals, using polymerase chain reaction(PCR) technology can detect serum and (or) liver tissue HBVDNA, called occult HBVinfection. This infection can occur in single or multiple antibody-positive, and can alsobe found in individuals with the hepatitis B virus serum markers (HBVM) werenegative.Occult HBV infection is also possible by blood transfusion, organtransplantation, hemodialysis, and other maternal and child vertical transmission of;will lead to a variety of harmful consequences, such as: HB immunization vaccinefailure, fulminant hepatic failure, chronic liver injury; HCV infection overlap increasethe severity of the disease; lead to the occurrence of hepatocellular carcinoma, etc.Objective: (1)to discusses HBVDNA in serum and PBMC of the unexplainedliver ill patients, judge the diagnosis of occult HBV infection significance with the twomethod;(2) study the gene variation with HBV S region of, to analysis molecularbiological mechanisms. of the occult HBV infection .Methods: 76 patients with unexplained liver disease, separating serum andPBMC, serum and PBMC were divided into two groups, the two groups wasamplified by nested PCR ,the positive analysis their clinical data, positive in serumanalysis their variable sites by sequencingHBV S gene .Results: (1) HBVDNA serum and PBMC were positive in 21.05% (16/76), and9.21% (7/76), the difference in the two groups was statistically significant, positive inserum group was higher than PBMC . (2) a case of serum HBVDNA is negative butwithin PBMC HBVDNA positive.(3) 7 patients with positive PBMC in HBVDNA,clinical diagnosis of both cirrhosis. Found that clinical symptoms, PBMC-positivecirrhosis of the liver higher risk of complications, severe illness, complications andmore. (4) 8 cases with positive in serum HBVDNA sequencing of were selectedrandomly, a determinant found within the sequence alignment, A→T at nt574mutation occurred, the deduced amino acids did not change; nt587 G→A mutationoccurs is derived from amino acid glycine mutation Arg, that is, the more commonG145R mutation. a determinant of foreign gene sequences found, nt355, nt484 baseby A→C, the deduced amino acids did not change, nt512 base by a C→A, deducedamino acid proline for the threonine variant.Conclusions: (1) for the diagnosis of occult HBV infection, serum is superior thatPBMC, but the detection of HBVDNA PBMC in serum can be a supplement. (2)PBMC occult HBV infection and cirrhosis in HBVDNA positive close relationshipwith judging the progress of the disease some guidance. (3) G145R mutation was oneof reason of occult HBV infection.