Restriction fragment length polymorphism analysis and direct sequencing for determination of HBV genotypes in a Turkish population.
The aim of this study was to analyze the restriction fragment length polymorphism and direct sequencing results in genotyping of hepatitis B virus from a Turkish population in a clinical virology laboratory. Serum samples of 54 chronic hepatitis B patients attending the Ege University Hospital were studied. Sequences of partial S gene PCR products were analysed and RFLP was performed. Fifty-three isolates could be identified by direct sequencing as genotype D. One sample needed to be cloned and determined as genotype D. Forty-two isolates were genotyped as D with RFLP according to published determinative patterns. Twelve isolates had undefined patterns. Eight of them suggested a mixture of isolates with different patterns and cloning of these samples confirmed the presence of heterogeneous isolates. Four isolates with undefined pattern were determined as genotype D by direct sequencing. All the studied isolates were genotype D. The results of this studied population suggest that RFLP is suitable for HBV genotyping in a routine clinical virology laboratory setting. However sequence analysis and even cloning may be needed to clarify indeterminate results.
Sertoz
Ruchan Yazan
RY
Ege University Medical Faculty, Department of Microbiology and Clinical Microbiology, Izmir, Turkey. ruchan.sertoz@ege.edu.tr
Erensoy
Selda
S
Pas
Suzan
S
Ozacar
Tijen
T
Niesters
Hubert
H
D016000:Cluster Analysis; D016172:DNA Fingerprinting; D004279:DNA, Viral / Q000235:genetics; D005838:Genotype; D006514:Hepatitis B Surface Antigens / Q000235:genetics; D006515:Hepatitis B virus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D019694:Hepatitis B, Chronic / Q000821:virology*; D006801:Humans; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA / Q000379:methods*; D014421:Turkey
D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens