[Results of a novel real-time PCR, sequence analysis, Inno-LiPA line probe assays in the detection of hepatitis B virus G1896A precore mutation in French blood donors].
OBJECTIVE
To screen hepatitis B virus (HBV) genotypes and associated basal core promoter (BCP; T1762A/A1764) and precore (PC; A1896) mutations among the 100 HBV surface antigen (HBsAg) positive voluntary blood donors in France.


METHODS
HBV genotypes were determined by using direct sequence analysis. Three methods were used to detect G1896A mutation: non-commercial real-time PCR (PCRTR°, line probe assay (InnoLiPA HBV PreCore, INNOGENETICS(®)) and direct sequencing of precore gene. HBV viral load was quantified with two commercial real-time PCR (COBAS(®) AmpliPrep/COBAS(®) TaqMan(®) HBV Test/Roche and Real Time HBV/M2000/Abbott).


RESULTS
The mean age of donors was 30 (18-64). Patients were from Africa (42%), Europa (50%), and Asia (8%). HBV/D was the most predominant (37%) genotype followed by HBV/A (31%) and HBV/E (22%). PC and BCP mutants were found in 57% with Inno-LIPA HBV test and 59% with both PCRTR and sequencing methods. A significant difference in the viral load of blood donors with wild and PC mutants was observed with the Taqman Cobas real time PCR (3,19 Log(10) UI/ml versus 4,93 Log(10) UI/ml, p < 0.05). Precore phenotype determination was in agreement with the three PC mutation detection methods in 56% of cases.


CONCLUSIONS
Non-Caucasian genotype E was present in the French blood donors. PC mutation was more common than BCP mutations in this study. As HBV infected blood donors were more often asymptomatic carriers, we could speculate that the G1896A mutation may favour the asymptomatic state, supporting previous observations.
Ducancelle
A
A
Laboratoire de virologie, CHU d'Angers, 4, rue Larrey, 49000 Angers, France. alducancelle@chu-angers.fr
Servant-Delmas
A
A
Beuvelet
T
T
Balan
V
V
Pivert
A
A
Maniez
M
M
Laperche
S
S
Lunel-Fabiani
F
F
D000293:Adolescent; D000328:Adult; D000349:Africa / Q000208:ethnology; D001208:Asia / Q000208:ethnology; D001782:Blood Donors; D003199:Computer Systems; D004252:DNA Mutational Analysis / Q000379:methods*; D005060:Europe / Q000208:ethnology; D005260:Female; D005602:France / Q000453:epidemiology; D005838:Genotype; D006509:Hepatitis B / Q000453:epidemiology / Q000235:genetics / Q000821:virology*; D006514:Hepatitis B Surface Antigens / Q000032:analysis; D006515:Hepatitis B virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D008403:Mass Screening; D008875:Middle Aged; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D017422:Sequence Analysis, DNA; D014766:Viremia / Q000453:epidemiology / Q000235:genetics / Q000821:virology*; D055815:Young Adult
D006514:Hepatitis B Surface Antigens; D011933:Reagent Kits, Diagnostic