Novel hiv-2 isolate
AssigneeTulane University Health Sciences Center
InventorsStephen M. Smith, Preston A. Marx, JR.
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Abstract
The invention provides a novel strain of HIV-2 capable of causing immunodeficiency. The invention also provides compositions comprising the nucleic acids and polypeptides characteristic of this HIV-2 virus, antibodies specific for this HIV-2 virus, methods of using these compositions, and methods of detecting HIV-2 virus.
Claims32
1-28. (canceled)
An isolated monoclonal antibody, or fragment thereof, that specifically binds to a human immunodeficiency virus type 2 (HIV-2) deposited as HIV-2NWK08F.
A method for the preparation of a hybridoma cell that produces monoclonal antibodies specific for an HIV-2NWK08F polypeptide, comprising the following steps:
- a. immunizing a mammal with an HIV-2NWK08F polypeptide selected from the group consisting of the amino acid sequence of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:6, or an immunogenic fragment thereof;
- b. isolating immunized splenocytes from said mammal;
- c. fusing the immunized splenocytes with a myeloma cell line to produce hybridomas;
- d. selecting for the hybridomas by culturing in selective media;
- e. clonally expanding the hybridomas in appropriate culture media;
- and,
- f. identifying and characterizing those hybridomas that produce monoclonal antibodies specific for an HIV-2NWK08F polypeptide.
A method for detecting HIV-2NWK08F antibodies in a biological sample comprising:
- a. contacting a biological sample with a composition comprising an HIV-2NWK08F polypeptide selected from the group consisting of the amino acid sequence of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, and SEQ ID NO:6, or a fragment thereof;
- and
- b. detecting an immunological complex formed between the HIV-2NWK08F polypeptide and the HIV-2NWK08F antibody.
A method of amplifying a target HIV-2NWK08F DNA sequence comprising:
- a. providing a reaction mixture comprising:
- i. a double-stranded target DNA comprising DNA isolated from a patient suspected of having HIV-2;
- ii. a primer pair comprising a first primer and a second primer, wherein the first primer is complementary to a sequence on the first strand of the target DNA and the second primer is complementary to a sequence on the second strand of the target DNA, and wherein the primer pair is capable of amplifying a DNA sequence within HIV-2NWK08F;
- iii.
- polymerase;
- and iv. free nucleotides comprising adenine, thymine, cytosine and guanine;
- b. heating and cooling the reaction mixture of (a) to amplify the target DNA;
- and
- c. analyzing the amplification activity.
A method of detecting HIV-2NWK08F nucleic acids in a biological sample comprising:
- a. contacting the biological sample with a primer or probe, wherein the primer or probe comprises at least 6, 8,10,12,15, 20, 25, 30, 40, or 50 consecutive nucleotides derived from the HIV-2NWK08F genome, and is no longer than about 50, 75, 100, or 200 nucleotides, and is capable of binding to a nucleic acid that encodes all or a portion of the amino acid sequence of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:6;
- and
- b. detecting the presence of nucleic acid that encodes all or a portion of the amino acid sequence of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:6.
A method of detecting a HIV-2NWK08F DNA in a test sample by polymerase chain reaction, the method comprising the steps of:
- amplifying a region consisting of:
- a. the nucleotides of SEQ ID NO:1;
- b. nucleotides 8-1417 of SEQ ID NO:1;
- c. nucleotides 1314-1417 of SEQ ID NO:1;
- d. the nucleotides of SEQ ID NO:4;
- e. nucleotides 1-827 SEQ ID NO:4;
- f. nucleotides 1046-2560 of SEQ ID NO:4;
- or
- g. nucleotides 2218-2263 of SEQ ID NO:4,
- using a nucleic acid molecule in the test sample or a nucleic acid molecule extracted from the test sample as a template with a primer pair capable of amplifying the region;
- and detecting the amplified region.
A primer pair comprising a first primer and a second primer, wherein the first and second primer each comprise at least 6, 8,10,12,15, 20, 25, 30, 40, or 50 consecutive nucleotides derived from the HIV -2NWK08F genome, and is no longer than about 50, 75, 100, or 200 nucleotides, wherein the primer is capable of binding to a nucleic acid that encodes all or a portion of the amino acid sequence of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:6.
A probe comprising at least 6, 8,10,12,15, 20, 25, 30, 40, or 50 consecutive nucleotides derived from the HIV-2NWK08F genome, wherein the probe is no longer than about 50, 75, 100, or 200 nucleotides, and wherein the probe is capable of binding to a nucleic acid that encodes all or a portion of the amino acid sequence of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:6, wherein the probe further comprises a detectable label.
Dependencies are inferred from the claim text.