# US 8,785,609 B1 · Cloning and expression of HTLV-III DNA

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- Number: US 8,785,609 B1
- Document ID: US8785609B1
- Status: Granted
- Kind: B1
- Application number: 08/462,504
- Filed: 1995-06-05
- Granted: 2014-07-22
- Earliest claimed: 1984-08-22
- Assignees: The United States of America, as represented by the Secretary, Department of Health and Human Services
- Applicants: Nancy T. Chang; Robert C. Gallo; Flossie Wong-Staal
- Inventors: Nancy T. Chang; Robert C. Gallo; Flossie Wong-Staal
- Web page: https://patentgrep.com/patent/US8785609B1

## CPC

- C12Q 1/702: {for retroviruses}
- C07K 14/005: from viruses
- C07K 16/1143: {Gag-pol proteins, e.g. p17or p24}
- C07K 16/1145: {Env proteins, e.g. gp41, gp110/120, gp160, V3, principal neutralising domain [PND] or CD4-binding site}
- C12Q 1/703: {Viruses associated with AIDS}
- A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00)
- C07K 2319/00: Fusion polypeptide
- C12N 2740/14022: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2740/16122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The determination of the nucleotide sequence of HTLV-III DNA; identification, isolation and expression of HTLV-III sequences which encode immunoreactive polypeptides by recombinant DNA methods and production of viral RNA are disclosed. Such polypeptides can be employed in immunoassays to detect HTLV-III.

## Claims

### Claim 1

A method of detecting a human immunodeficiency virus (HIV) nucleic acid in a fluid sample, the method comprising the steps of:
(a) contacting the fluid sample with a nucleic acid probe comprising an HIV nucleotide sequence complementary to that depicted in FIG. 3, or a portion thereof, wherein an HIV nucleic acid, if present in the fluid sample, specifically hybridizes to a nucleic acid complementary to the nucleotide sequence of FIG. 3; and
(b) determining duplex formation between the nucleic acid probe and the HIV nucleic acid, if present in the fluid sample,
wherein the nucleic acid probe forms a duplex with an HIV nucleic, but does not form a duplex with an HTLV-I nucleic acid or an HTLV-II nucleic acid.

### Claim 2

A method comprising the steps of:
(a) providing a fluid sample suspected of containing a human immunodeficiency virus (HIV) nucleic acid;
(b) providing a nucleic acid comprising a nucleotide sequence specific to HIV comprising an HIV nucleotide sequence complementary to that depicted in FIG. 3, or a portion thereof; and
(c) combining nucleic acids in the fluid sample and the nucleic acid of step (b) under hybridization conditions that permit duplex formation between the nucleic acid of step (b) and the HIV nucleic acid but not between the nucleic acid of step (b) and an HTLV-I nucleic acid or an HTLV-II nucleic acid,
wherein, if the nucleic acid of step (b) forms a duplex with a nucleic acid in the sample, the HIV nucleic acid is detected in the sample, and wherein, if the nucleic acid of step (b) does not form a duplex with a nucleic acid in the sample, the HIV nucleic acid is not detected in the sample.

### Claim 3

A method comprising the step of:
combining a fluid sample suspected of containing a human immunodeficiency virus (HIV) nucleic acid with a nucleic acid comprising a nucleotide sequence specific for HIV selected from the group consisting of:
(i) a nucleic acid comprising an HIV nucleotide sequence as depicted in FIG. 3, or a portion thereof, and
(ii) a nucleic acid comprising an HIV nucleotide sequence complementary to the nucleotide sequence as depicted in FIG. 3, or a portion thereof;
wherein the combining of the fluid sample and the nucleic acid is under hybridization conditions that permit duplex formation between the nucleic acid of (i) or (ii) and an HIV nucleic acid in the fluid sample but not between the nucleic acid of (i) or (ii) and an HTLV-I or an HTLV-II nucleic acid,
wherein, if the nucleic acid of (i) or (ii) forms a duplex with a nucleic acid in the fluid sample, the HIV nucleic acid is detected in the fluid sample, and wherein, if the nucleic acid of (i) or (ii) does not form a duplex with a nucleic acid in the fluid sample, the HIV nucleic acid is not detected in the fluid sample.

### Claim 4

A method of detecting a human immunodeficiency virus (HIV) nucleic acid in a bodily fluid, the method comprising the steps of:
(a) contacting the bodily fluid with a nucleic acid selected from the group consisting of:
(i) a nucleic acid comprising an HIV nucleotide sequence as depicted in FIG. 3, or a portion thereof, and
(ii) a nucleic acid comprising an HIV nucleotide sequence complementary to the nucleotide sequence as depicted in FIG. 3, or a portion thereof;
and
(b) determining duplex formation between the nucleic acid of (i) or (ii) and an HIV nucleic acid, if present in the bodily fluid,
wherein the nucleic acid of (i) or (ii) forms a duplex with an HIV nucleic acid, but does not form a duplex with an HTLV-I nucleic acid or an HTLV-II nucleic acid.

### Claim 5

A method of detecting a human immunodeficiency virus (HIV) nucleic acid in a nucleic acid sample, the method comprising the steps of:
(a) contacting a nucleic acid sample suspected of containing an HIV nucleic acid with a nucleic acid selected from the group consisting of:
(i) a nucleic acid comprising an HIV nucleotide sequence as depicted in FIG. 3, or a portion thereof, and
(ii) a nucleic acid comprising an HIV nucleotide sequence complementary to the nucleotide sequence as depicted in FIG. 3, or a portion thereof;
and
(b) determining whether or not the nucleic acid of (i) or (ii) hybridizes to an HIV nucleic acid in the sample,
wherein the nucleic acid of (i) or (ii) forms a duplex with an HIV nucleic acid, but does not form a duplex with an HTLV-I nucleic acid or an HTLV-II nucleic acid,
wherein, if the nucleic acid of (i) or (ii) hybridizes to the HIV nucleic acid in the nucleic acid sample, the HIV nucleic acid is detected in the nucleic acid sample, and
wherein, if the nucleic acid of (i) or (ii) does not hybridize to a nucleic acid in the nucleic acid sample, the HIV nucleic acid is not detected in the nucleic acid sample.

### Claim 6 (depends on 1)

The method according to claim 1, wherein the nucleic acid probe comprises a nucleotide sequence complementary to a gag, pol, or env-lor open reading frame or a LTR region.

### Claim 7 (depends on 6)

The method according to claim 6, wherein the open reading frame is the gag open reading frame and wherein the gag open reading frame extends from nucleotide 334 to nucleotide 1870 of FIG. 3.

### Claim 8 (depends on 6)

The method according to claim 6, wherein the open reading frame is the pol open reading frame and wherein the pol open reading frame extends from nucleotide 1629 to nucleotide 4673 of FIG. 3.

### Claim 9 (depends on 6)

The method according to claim 6, wherein the open reading frame is the env-lor open reading frame and wherein the env-lor open reading frame extends from nucleotide 5781 to nucleotide 8369 of FIG. 3.

### Claim 10 (depends on 6)

The method according to claim 6, wherein the nucleic acid probe is from within nucleotide 3554 to nucleotide 6664 as depicted in FIG. 3.

### Claim 11 (depends on 1)

The method according to claim 1, wherein the nucleic acid probe comprises RNA.

### Claim 12 (depends on 1)

The method according to claim 1, wherein the nucleic acid probe comprises DNA.

### Claim 13 (depends on 1)

The method according to claim 1, wherein the nucleic acid probe comprises a label.

### Claim 14 (depends on 2, 3, 4, 5)

The method as in one of claims 2-5, wherein the nucleic acid comprises a nucleotide sequence complementary to a gag, pol, or env-lor open reading frame or a LTR region.

### Claim 15 (depends on 14)

The method according to claim 14, wherein the open reading frame is the gag open reading frame and wherein the gag open reading frame extends from nucleotide 334 to nucleotide 1870 of FIG. 3.

### Claim 16 (depends on 14)

The method according to claim 14, wherein the open reading frame is the pol open reading frame and wherein the pol open reading frame extends from nucleotide 1629 to nucleotide 4673 of FIG. 3.

### Claim 17 (depends on 14)

The method according to claim 14, wherein the open reading frame is the env-lor open reading frame and wherein the env-lor open reading frame extends from nucleotide 5781 to nucleotide 8369 of FIG. 3.

### Claim 18 (depends on 14)

The method according to claim 14, wherein the nucleic acid of is from within nucleotide 3554 to nucleotide 6664 as depicted in FIG. 3.

### Claim 19 (depends on 2, 3, 4, 5)

The method as in one of claims 2-5, wherein the nucleic acid comprises RNA.

### Claim 20 (depends on 2, 3, 4, 5)

The method as in one of claims 2-5, wherein the nucleic acid comprises DNA.

### Claim 21 (depends on 2, 3, 4, 5)

The method as in one of claims 2-5, wherein the nucleic acid comprises a label.