Comparative methods for genotyping hepatitis C virus isolates from Romania.
Accurate genotyping of hepatitis C virus (HCV) has clinical implications for treatment orientation and epidemiological impact in tracing the contamination sources. The aim of the study was to compare a genotyping assay by restriction fragment length polymorphism (RFLP) in the HCV 5'untranslated region (5'UTR) with sequencing in the 5'untranslated and NS5B regions. One hundred and three samples, collected between 2004 and 2006 from chronically infected patients with HCV, were tested with the 5'UTR and NS5B protocols. Of the total number of the samples tested by the 5'UTR-RFLP assay (n=103) the HCV subtype could be inferred by this method for 92 samples, by 5'UTR sequencing for 16 samples out of 23 tested (n=23) and by using the NS5B sequencing for all the samples tested (n=34). Our results showed that the HCV genotype distribution in Romania is: 1b--86.4%, 1a--10.7% and 4a--2.9%. In conclusion, RFLP screening in the 5'UTR is a convenient method for HCV genotyping and discrimination between 1b and non-1b genotypes but has a poor resolving power for subtyping and evaluation of the transmission routes. Sequencing in NS5B region is more adapted than RFLP and sequencing in 5'UTR for subtyping and epidemiological investigation.
Oprişan
Gabriela
G
NIRDMI Cantacuzino, Bucharest, Romania. goprisan@cantacuzino.ro
Szmal
Camelia
C
Dinu
Sorin
S
Oprişoreanu
Ana-Maria
AM
Thiers
Valérie
V
Panait
Mircea
M
Oţelea
Dan
D
Mavromara
Penelope
P
Ruţă
Simona
S
Sultana
Camelia
C
Alexiu
Irina
I
Manolescu
Loredana
L
Anton
Gabriela
G
Grancea
Camelia
C
Neagu
Ana
A
Sencovici
Carmen
C
Calistru
Petre Jacob
PJ
Târdei
Gratiela
G
Moţoc
Adriana
A
Lazăr
Stefan
S
Ionescu
Camelia
C
Ceauşu
Emanoil
E
Cristea
Cristiana
C
Voiculescu
Ghe
G
Brehar-Cioflec
Dana
D
Popovici
Damian
D
Chicin
Gratiana
G
Claici
Camelia
C
D020121:5' Untranslated Regions; D005838:Genotype; D016174:Hepacivirus / Q000235:genetics* / Q000302:isolation & purification; D006521:Hepatitis, Chronic / Q000097:blood / Q000821:virology*; D006801:Humans; D010802:Phylogeny; D012150:Polymorphism, Restriction Fragment Length; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012383:Romania; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics; D012324:RNA-Dependent RNA Polymerase
D020121:5' Untranslated Regions; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase