Compositions, kits, and methods to detect HIV virus
Provided herein include compositions comprising a primer having the nucleic acid sequence of ACeIN-F3_c, a primer having the nucleic acid sequence of ACeIN-B3_a, a primer having the nucleic acid sequence of ACeIN-B3_b, a primer having the nucleic acid sequence of ACeIN-FIP_e, a primer having the nucleic acid sequence of ACeIN-FIP_f, a primer having the nucleic acid sequence of ACeIN-BIP (or ACeIN-BIP-song), a primer having the nucleic acid sequence of ACeIN-LF; and a primer having the nucleic acid sequence of ACeIN-LB. Also provided are methods of detecting human immunodeficiency virus (HIV) nucleic acids in a sample comprising performing reverse transcription-based loop mediated isothermal amplification (RT-LAMP) on a sample using the previously disclosed compositions.
1. A composition, comprising:
a primer comprising SEQ ID NO: 37;
a primer comprising SEQ ID NO: 39;
a primer comprising SEQ ID NO: 41;
a primer comprising SEQ ID NO: 47;
a primer comprising SEQ ID NO: 50;
a primer comprising SEQ ID NO: 63;
a primer comprising SEQ ID NO: 67; and
a primer comprising SEQ ID NO: 70.
2. A composition, comprising:
a primer comprising SEQ ID NO: 37;
a primer comprising SEQ ID NO: 39;
a primer comprising SEQ ID NO: 41;
a primer comprising SEQ ID NO: 47;
a primer comprising SEQ ID NO: 50;
a primer comprising SEQ ID NO: 64;
a primer comprising SEQ ID NO: 67; and
a primer comprising SEQ ID NO: 70.
3. A method of detecting human immunodeficiency virus (HIV) nucleic acids in a sample from a patient comprising:
providing a reaction mixture comprising a first primer set or a second primer set for performing a reverse transcription-based loop mediated isothermal amplification assay (RT-LAMP) assay, magnesium, dNTPs, a reaction buffer, a DNA polymerase and a sample to be tested for presence of HIV nucleic acids;
wherein the first primer set comprises:
a primer comprising SEQ ID NO: 37;
a primer comprising SEQ ID NO: 39;
a primer comprising SEQ ID NO: 41;
a primer comprising SEQ ID NO: 47;
a primer comprising SEQ ID NO: 50;
a primer comprising SEQ ID NO: 63;
a primer comprising SEQ ID NO: 67; and
a primer comprising SEQ ID NO: 70; and wherein the second primer set comprises:
a primer comprising SEQ ID NO: 37;
a primer comprising SEQ ID NO: 39;
a primer comprising SEQ ID NO: 41;
a primer comprising SEQ ID NO: 47;
a primer comprising SEQ ID NO: 50;
a primer comprising SEQ ID NO: 64;
a primer comprising SEQ ID NO: 67; and
a primer comprising SEQ ID NO: 70;
incubating the reaction mixture under polymerase reaction conditions so as to produce a reaction product comprising amplified HIV nucleic acids; and detecting the reaction product, wherein the detection has a Z-factor of above 0.5.
4. The method of claim 3 , wherein the sample to be tested for presence of HIV nucleic acids comprises one or more subtypes selected from the group consisting of subtype A, B, C, D, and G.
5. The method of claim 3 , wherein the detected reaction product is a human immunodeficiency virus nucleic acid of subtype A, B, C, D, or G.
6. The method of claim 4 , wherein the detected reaction product is a human immunodeficiency virus nucleic acid of subtype B.
7. The method of claim 3 , wherein the detected reaction product comprises HIV integrase coding region of pol gene.
8. The method of claim 3 , wherein the sample is a neonate sample comprising maternal antibodies.
9. The method of claim 3 , wherein the patient has not undergone seroconversion.
10. A method of monitoring a response to a medication for HIV infection in a subject in need thereof, comprising:
obtaining a first sample from the subject at a first time point before administering the medication for HIV infection to the subject:
obtaining a second sample from the subject at a second time point after administering the medication for HIV infection to the subject;
determining the amount of human immunodeficiency virus (HIV) in the first and second samples, the determining comprising:
performing a reverse transcription-based loop mediated isothermal amplification (RT-LAMP) on a sample containing HIV using either a first primer set for the first and second samples or a second primer set for the first and second samples, wherein the first primer set comprises:
a primer comprising SEQ ID NO: 37;
a primer comprising SEQ ID NO: 39;
a primer comprising SEQ ID NO: 41;
a primer comprising SEQ ID NO: 47;
a primer comprising SEQ ID NO: 50;
a primer comprising SEQ ID NO: 63;
a primer comprising SEQ ID NO: 67; and
a primer comprising SEQ ID NO: 70;
wherein the second primer set comprises:
a primer comprising SEQ ID NO: 37;
a primer comprising SEQ ID NO: 39;
a primer comprising SEQ ID NO: 41;
a primer comprising SEQ ID NO: 47;
a primer comprising SEQ ID NO: 50;
a primer comprising SEQ ID NO: 64;
a primer comprising SEQ ID NO: 67; and
a primer comprising SEQ ID NO: 70; and
comparing the amount of HIV in the first and second samples,
wherein a decrease in the amount of HIV from the first sample relative to the second sample indicates a positive response of the subject to the medication treatment of HIV infection.
11. The method of claim 10 , wherein the medication is an antiretroviral therapy.
12. The method of claim 10 , wherein the sample comprises aqueous humour, vitreous humour, bile, blood, blood serum, breast milk, cerebrospinal fluid, endolymph, perilymph gastric juice, mucus, peritoneal fluid, pleural fluid, saliva, sebum, semen, sweat, tears, vaginal secretion, vomit, or urine.
13. The method of claim 10 , wherein the first and/or second sample(s) comprise(s) about 50, about 500, about 1000, about 5000 or about 5500 copies of HIV.
14. The method of claim 3 , wherein the sample comprises aqueous humour, vitreous humour, bile, blood, blood serum, breast milk, cerebrospinal fluid, endolymph, perilymph gastric juice, mucus, peritoneal fluid, pleural fluid, saliva, sebum, semen, sweat, tears, vaginal secretion, vomit, or urine.
15. The method of claim 3 , wherein the sample comprises about 50, about 500, about 1000, about 5000 or about 5500 copies of HIV.