Naturally occurring mutations associated with resistance to HCV NS5B polymerase and NS3 protease inhibitors in treatment-naïve patients with chronic hepatitis C

Table 1 Primers used for HCV NS3 protease gene amplification

HCV genotype	I-PCR primers	II-PCR primers	5’-3’ sequence^b	H77 location
1b	MarsF2^a MarsR1^a	NS31b F^b MarsR2^a	5'- TRCCHGTCTCCGCCCGVAG-3'^c	3291-3310 4215–4237 3340–3358 4035-4054
2c	MarsF1^a MarsR1^a	NS3 2c F^b NS3 2c R^b	5'- TGGGCCCTGCTGATGGATAC-3'^d 5'- GTAGGTCTGGGGCACAGCTG-3'	3291-3310 4215–4237 3376–3395 4010-3991
4d	G4F1^a G4R1^a	NS3 4d F^b NS3 4d R^b	5’-ATGCGCACRCYAYGAAGGG-3’^e 5’-GGCACGGCAGGAGGAGTGGA-3’	3369-3388 3997–4018 3388–3407 4000-3981

^a Previously published primers (Vallet S [30])
^b Primers selected by alignment of sequences from Los Alamos databank
^c Amplification conditions in the second PCR for genotype 1b, were: 2’ at 94 °C, and then 30 cycles at 94 °C for 30”, 60 °C for 30” and 72 °C for 45”, plus extension at 72 °C for 7’
^d Amplification conditions in the second PCR for genotype 2c, were: 2’at 94 °C, and then 30 cycles at 94 °C for 30”, 62 °C for 30” and 72 °C for 45”, plus extension at 72 °C for 7’
^e Amplification conditions in the second PCR for genotype 4, were: 2’at 94 °C, and then 30 cycles at 94 °C for 30”, 60 °C for 40” and 72 °C for 45”, plus extension at 72 °C for 7’

ISSN: 1743-422X