IP Library › Granted Patent US 12,492,437
Granted Patent B2
US 12,492,437 · App. 17/850,480 · Granted Dec 9, 2025

Compositions and methods for detecting or quantifying hepatitis C virus

Inventors: Siobhan Miick (San Diego, CA); Paul M. Darby (San Diego, CA); Jo Ann Jackson (Lakeside, CA); Sheila M.J. Aubin (San Diego, CA)
Assignee: Gen-Probe Incorporated
C12Q1/706C12Q1/6806C12Q1/6876C12Q1/686C12Q2525/113C12Q2525/125C12Q2525/143C12Q2525/301C12Q2527/143C12Q2600/16
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Quick Facts
Patent No.
US 12,492,437
App. No.
17/850,480
Granted
Dec 9, 2025
Kind
B2
Abstract

This disclosure provides oligomers, compositions, and kits for detecting and quantifying Hepatitis C virus (HCV), including different genotypes and variants thereof, and related methods and uses. In some embodiments, oligomers target the 5′ untranslated region of HCV and are configured to provide substantially equivalent quantification of different genotypes and variants of HCV.

Claims (28)

1 . A method of detecting Hepatitis C virus in a sample comprising:

contacting the sample with at least first, second, and third amplification oligomers, thereby forming a composition,

performing a nucleic acid amplification reaction in the composition which produces one or more amplicons in the presence of a Hepatitis C virus nucleic acid, and

detecting the amplicon, wherein:

the first amplification oligomer comprises a target-hybridizing sequence comprising at least 10 contiguous nucleotides of SEQ ID NO: 2, including at least a portion of the nucleotide sequence containing positions 5, 7, 12, and 15 of SEQ ID NO: 2; and

the second amplification oligomer comprises a target-hybridizing sequence comprising at least 10 contiguous nucleotides of SEQ ID NO: 3 including at least a portion of the nucleotide sequence containing positions 5, 7, 12, and 15 of SEQ ID NO: 3; and

the third amplification oligomer comprises at least about 14 contiguous nucleotides of antisense Hepatitis C virus sequence and is configured to specifically hybridize downstream of HCV genomic position 78 of SEQ ID NO: 75; and

the target-hybridizing sequences of the first and second amplification oligomers each comprise at least about 14 contiguous nucleotides of Hepatitis C virus sequence; and

the one or more amplicons are produced through extension of the first and third amplification oligomers or the second and third amplification oligomers in the presence of the Hepatitis C virus nucleic acid; and (i) at least one of the amplification oligomers is a promoter-primer; or (ii) the one or more amplicons are detected with a probe oligomer, wherein the probe oligomer comprises a non-nucleotide detectable label and/or wherein at least about half of the sugar moieties in the probe oligomer are 2′-O-methyl-ribose.

2 . The method of claim 1 , wherein the first amplification oligomer comprises a target-hybridizing sequence comprising at least one, two, three, or four of SEQ ID NOs: 23-27; or wherein the first amplification oligomer comprises at least 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26 contiguous nucleotides of SEQ ID NO: 2; or wherein the first amplification oligomer comprises the sequence of SEQ ID NO: 2.

3 . The method of claim 1 , wherein the second amplification oligomer comprises a target-hybridizing sequence comprising at least one, two, three, or four of SEQ ID NOs: 28-32; or wherein the second amplification oligomer comprises at least 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26 contiguous nucleotides of SEQ ID NO: 3; or wherein the second amplification oligomer comprises the sequence of SEQ ID NO: 3.

4 . The method of claim 1 , wherein the third amplification oligomer does not anneal downstream of an HCV genomic position selected from a position in at least one HCV type that corresponds to position 120, 125, 130, 135, 140, 145, or 150 of SEQ ID NO: 75.

5 . The method of claim 4 , wherein the at least one HCV type includes one or more of HCV types 1a, 1b, 2b, 3b, 4b, 5a, and 6a.

6 . The method of claim 1 , wherein the third amplification oligomer is configured to specifically hybridize to a site comprising at least a portion of the nucleotide sequence containing positions 80-119 of SEQ ID NO: 75; or wherein the third amplification oligomer comprises a target-hybridizing sequence comprising at least 10 contiguous nucleotides of SEQ ID NO: 6 or 7; or wherein the third amplification oligomer comprises a target-hybridizing sequence comprising at least one, two, three, or four of SEQ ID NOs: 33-37; or wherein the third amplification oligomer comprises 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26 contiguous nucleotides of SEQ ID NO: 7; or wherein the third amplification oligomer comprises the sequence of SEQ ID NO: 7; or wherein the third amplification oligomer comprises the sequence of at least one, two, three, four, or five of SEQ ID NOs: 42-47; or wherein the third amplification oligomer comprises the sequence of SEQ ID NO: 5.

7 . The method of claim 1 , wherein the first and second amplification oligomers are present in relative molar amounts (first: second) ranging from about 8.5:1.5 to about 1.5:8.5, about 7.5:2.5 to about 2.5:7.5, about 8:2 to about 7:3, about 7:3 to about 6:4, about 6:4 to about 5:5, about 5:5 to about 4:6, about 4:6 to about 3:7, or about 3:7 to about 2:8; or wherein the first and second amplification oligomers are present in relative molar amounts (first: second) ranging from about 6:4 to about 1.5:8.5, about 4:6 to about 6:4, or about 4.5:5.5 to about 5.5:4.5.

8 . The method of claim 1 , further comprising contacting the sample with an initial amplification oligomer, wherein the initial amplification oligomer comprises a target-hybridizing sequence comprising at least one, two, three, four, five, six, or seven of SEQ ID NOs: 33-41; or wherein the initial amplification oligomer comprises 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39 contiguous nucleotides of SEQ ID NO: 6; or wherein the initial amplification oligomer comprises the sequence of SEQ ID NO: 6; or wherein the initial amplification oligomer comprises the sequence of at least one, two, three, four, or five of SEQ ID NOs: 42-47; or wherein the initial amplification oligomer comprises the sequence of SEQ ID NO: 4.

9 . The method of claim 1 , wherein the probe oligomer comprises a target-hybridizing sequence comprising at least one or two of SEQ ID NOs: 50-52; or wherein the probe oligomer comprises the sequence of SEQ ID NO: 48 or 49; or wherein the probe oligomer comprises at least 11, 12, 13, 14, or 15 contiguous nucleotides of SEQ ID NO: 12; or wherein the probe oligomer comprises a target-hybridizing sequence comprising at least 11, 12, 13, 14, or 15 contiguous nucleotides of SEQ ID NO: 13; or wherein the probe oligomer comprises a first self-complementary region at its 5′ end and a second self-complementary region at its 3′ end; or wherein the probe oligomer comprises a first self-complementary region at its 5′ end and a second self-complementary region at its 3′ end and wherein the self-complementary regions can hybridize to form about 4 to 7 Watson-Crick or wobble base pairs; or wherein the probe oligomer comprises a first self-complementary region at its 5′ end and a second self-complementary region at its 3′ end and wherein the self-complementary regions can hybridize to form about 5 Watson-Crick or wobble base pairs; or wherein the probe oligomer comprises the sequence of SEQ ID NO: 12; or wherein the probe oligomer comprises a target-hybridizing sequence comprising the sequence of SEQ ID NO: 13.

10 . The method of claim 9 , wherein the probe oligomer comprises a non-nucleotide detectable label.

11 . The method of claim 10 , wherein the non-nucleotide detectable label is a fluorescent label.

12 . The method of claim 11 , wherein the probe oligomer comprises a quencher.

13 . The method of claim 12 , wherein the non-nucleotide detectable label is a fluorescent label and the quencher absorbs fluorescence to a greater extent when the probe is free than when the probe is annealed to a target nucleic acid.

14 . The method of claim 13 , wherein the fluorescent label is fluorescein, hexachlorofluorescein, carboxyrhodamine, or acridine.

15 . The method of claim 14 , wherein the quencher is DABCYL.

16 . The method of claim 15 , wherein the fluorescent label is attached to the 5′-terminus of the probe oligomer and the quencher is attached to the 3′-terminus of the probe oligomer, or the fluorescent label is attached to the 3′-terminus of the probe oligomer and the quencher is attached to the 5′-terminus of the probe oligomer.

17 . The method of claim 9 , wherein at least about half, at least about 90%, or all of the sugar moieties in the probe oligomer are 2′-O-methyl-ribose.

18 . The method of claim 1 , wherein at least one amplification oligomer is a promoter-primer comprising the sequence of SEQ ID NO: 8, 9, 10, or 11.

19 . The method of claim 1 , wherein the third amplification oligomer is configured to specifically hybridize downstream of position 78 of SEQ ID NO: 75 and is a promoter-primer.

20 . The method of claim 1 , wherein at least one amplification oligomer is a promoter-primer comprising a T7 promoter located 5′ of the target-hybridizing sequence.

Assignments (3)
RELEASE OF SECURITY INTEREST RECORDED AT REEL/FRAME 061639/0513 Recorded Apr 24, 2026
From: BANK OF AMERICA, N.A., AS COLLATERAL AGENT
To: HOLOGIC, INC.; GEN-PROBE INCORPORATED; FAXITRON BIOPTICS, LLC; BIOTHERANOSTICS, INC.
Reel/Frame 075457/0268 →
SECURITY INTEREST Recorded Apr 8, 2026
From: BIOTHERANOSTICS, INC.; GEN-PROBE INCORPORATED; GEN-PROBE PRODESSE, INC.; CYTYC CORPORATION; SUROS SURGICAL SYSTEMS, INC.; GYNESONICS, INC.; BOLDER SURGICAL, LLC; FAXITRON BIOPTICS, LLC; HEALTH BEACONS, INC.; HOLOGIC, INC.
To: ROYAL BANK OF CANADA, AS COLLATERAL AGENT
Reel/Frame 075462/0440 →
SECURITY INTEREST Recorded Oct 11, 2022
From: HOLOGIC, INC.; FAXITRON BIOPTICS, LLC; BIOTHERANOSTICS, INC.; GEN-PROBE INCORPORATED
To: BANK OF AMERICA, N.A., AS COLLATERAL AGENT
Reel/Frame 061639/0513 →
Continuity (3)
Division 15787344 · Oct 18, 2017
Provisional Application 62410188 · Oct 19, 2016
Related Publication 20220389525A1 · Dec 8, 2022
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