New combination test for hepatitis C virus genotype and viral load determination using Amplicor GT HCV MONITOR test v2.0

Table 1 Genotype specific capture probes (Aminolink1) and competitive oligonucleotide probes.

Probe name	Sequences	Use
MH1Ami	5'-Aminolink-tttttgctcaatgcctggagatttg-3'	Genotype 1 specific capture probe
MHCG1C	5'-cactctatgcccggcc-3'	Competitor oligonucleotide to genotype 2
MHCG1C3	5'-tcaatrcccraaatttg-3'	Competitor oligonucleotide to genotype 3
MH2Ami	5'- Aminolink-tttttcttggataaacccactctatgyc-3'	Genotype 2 specific capture probe
MHCG2C	5'-ggatcaacccgctcaatg-3'	Competitive oligonucleotide to genotype 1
MHCG2C3	5'-arcaacccgctcaatg-3'	Competitive oligonucleotide to genotype 3
MH3Ami	5'- Aminolink-tttttgaatcgctggggtgaccg-3'	Genotype 3 specific capture probe
MH3’Ami	5'- Aminolink-tttttcgcgagatcactagccgagtag-3'	Genotype 3 specific capture probe
MHCG3C	5'-aattgccrggamgac-3'	Competitive oligonucleotide to genotype 1, 2
MHCG3’C	5'-cgcragactgctagccga-3'	Competitive oligonucleotide to genotype 1, 2

¹Aminolink is a registered trademark of applied biosystems.

Table 2 Universal biotin-labeled HCV specific probes for the amplification of the detection signal.

Probe name	Sequences
MH211B+	5'-Biotin-ctagccgagtagcgttgggttgcgaaagg-3'
MH111B+	5'-Biotin-ctagccgagtagtgttgggtcgcgaaagg-3'
MHBT1	5'-Biotin-tagtatgagtgtcgtgcagcctccaggc-3'
MHBT2	5'-Biotin-gagagccatagtggtctgcggaaccggt-3'
MHBT3	5'-Biotin-gagtacaccggaattgccaggacgacc-3'
MHBT4	5'-Biotin ccttgtggtactgcctgatagggtgcttg-3'

Table 3 Detection limit of the HCV Guideline test in 264 samples infected with HCV.

Amplicor monitor	Number (IU/mL)	HCV-Guideline positivity, n (%)
<500	12	0 (0)
500-1000	18	18 (100.0)
1000-10000	73	73 (100.0)
10000-100000	71	71 (100.0)
100000-850000	62	62 (100.0)
850000<	28	28 (100.0)

Table 4 HCV genotypes of 252 serum samples from patients with chronic HCV infection as determined by HCV-Guideline test and a core genotyping method reported by Ohno and colleagues[19].

Core genotyping	HCV-Guideline test
	1	2	3	1+2	1+3	Total
1a	1	0	0	0	0	1
1b	147	0	0	0	0	147
2a	0	64	0	0	0	64
2b	0	15	0	0	0	15
3a	0	0	13	0	0	13
3b	0	0	1	0	0	1
2a+2b	0	2	0	0	0	2
1b+2a	2	1	0	0	0	3
1b+2b	0	0	0	1	0	1
1b+3a	0	0	1	0	1	2
Not classified	0	3	0	0	0	3
Total	150	85	15	1	1	252

Table 5 HCV genotypes and serotypes of 252 serum samples from patients with chronic HCV infection as determined by HCV-Guideline test and serological Immucheck HCV GR test.

Serotyping	HCV-Guideline test
	1	2	3	1+2	1+3	Total
G1	142	0	0	0	1	143
G2	11	76	0	0	0	77
Not determined	7	9	15	1	0	32
Total	150	85	15	1	1	252

¹HCV core sequence by direct sequencing indicated genotype 1b, and confirmed by a core genotyping method[19].