In vitro method for the detection and quantification of HIV-2

Abstract

The present invention relates to a method for detecting or quantifying Human Immunodeficiency Virus-2 (HIV-2) nucleic acids in a biological sample, comprising: a) performing a real-time polymerase chain reaction (PCR) or a real-time reverse transcriptase polymerase chain reaction (RT-PCR) on nucleic acids of the biological sample with: (i) at least 4 primers respectively comprising or consisting of: sequence SEQ ID NO: 1 or a sequence having at least 90% identity to SEQ ID NO 1, and sequence SEQ ID NO: 2 or a sequence having at least 90% identity to SEQ ID NO: 2, and sequence SEQ ID NO: 4 or a sequence having at least 90% identity to SEQ ID NO: 4, and sequence SEQ ID NO: 5 or a sequence having at least 90% identity to SEQ ID NO: 5, and (ii) at least 2 labelled probes respectively comprising or consisting of: sequence SEQ ID NO: 3, a sequence complementary to SEQ ID NO: 3, or a sequence having at least 90% identity to SEQ ID NO: 3 or the complementary thereof, and sequence SEQ ID NO: 6, a sequence complementary to SEQ ID NO: 6, or a sequence having at least 90% identity to SEQ ID NO: 6 or the complementary thereof, and b) determining therefrom the presence or absence and/or the quantity of HIV-2 nucleic acids in the biological sample.

The invention claimed is: 
     
       1. A method for detecting or quantifying Human Immunodeficiency Virus-2(HIV-2) nucleic acids in a biological sample, comprising:
 a) performing a real-time polymerase chain reaction (PCR) or a real-time reverse transcriptase polymerase chain reaction (RT-PCR) on nucleic acids of the biological sample with:
 (i) at least 4 primers respectively consisting of:
 sequence SEQ ID NO: 1, 
 sequence SEQ ID NO: 2, 
 sequence SEQ ID NO: 4, and 
 sequence SEQ ID NO: 5, and 
 
 (ii) at least 2 labelled probes respectively consisting of:
 sequence SEQ ID NO: 3, and 
 sequence SEQ ID NO: 6, and 
 
 
 b) determining therefrom the presence or absence and/or the quantity of HIV-2 nucleic acids in the biological sample. 
 
     
     
       2. The method according to  claim 1 , for detecting or quantifying HIV-2 RNA. 
     
     
       3. The method according to  claim 1 , wherein the labelled probes are labelled with 6-carboxyfluorescein (FAM) at their 5′ end and with Black Hole Quencher-1 (BHQ1) at their 3′ end. 
     
     
       4. The method according to  claim 1 , wherein the PCR or RT-PCR comprises
 the following thermocycling conditions:
 optionally 10 min at 50° C. for reverse transcription, followed by 
 5 min at 95° C., followed by 
 50 cycles of 95° C. for 15 s and 60° C. for 1 min. 
 
 
     
     
       5. The method according to  claim 1 , further comprising determining or quantifying HIV-1 nucleic acids in a biological sample. 
     
     
       6. A kit or a mix for detecting or quantifying HIV-2 nucleic acids, comprising:
 a) at least 4 primers respectively consisting of:
 sequence SEQ ID NO: 1, 
 sequence SEQ ID NO: 2, 
 sequence SEQ ID NO: 4, and 
 sequence SEQ ID NO: 5, and 
 
 b) at least 2 labelled probes respectively consisting of:
 sequence SEQ ID NO: 3, and 
 sequence SEQ ID NO: 6, and 
 
 c) optionally additional reagents for performing PCR or RT-PCR. 
 
     
     
       7. The kit or the mix according to  claim 6 , wherein the labelled probes are labelled with 6-carboxyfluorescein (FAM) at their 5′ end and with Black Hole Quencher-1 (BHQ1) at their 3′ end. 
     
     
       8. The kit or the mix according to  claim 6 , further comprising primers and labelled probes for detecting or quantifying HIV-1 nucleic acids. 
     
     
       9. An in vitro method for diagnosing HIV-2 infection, or determining HIV-2 viral load, in an individual, comprising:
 (a) performing the method for detecting or quantifying Human Immunodeficiency Virus-2 (HIV-2) nucleic acids in a biological sample taken from the individual as defined in  claim 1 ; 
 (b) determining therefrom whether the individual is infected by HIV-2 or the HIV-2 viral load of the individual. 
 
     
     
       10. An in vitro method for diagnosing HIV-2 infection, or determining HIV-2 viral load, in an individual, comprising the steps of:
 (a) performing the method for detecting or quantifying Human Immunodeficiency Virus-2 (HIV-2) nucleic acids in a biological sample taken from the individual as defined in  claim 3 ; 
 (b) determining therefrom whether the individual is infected by HIV-2 or the HIV-2 viral load of the individual. 
 
     
     
       11. An in vitro method for diagnosing HIV-2 infection, or determining HIV-2 viral load, in an individual, comprising the steps of:
 (a) performing the method for detecting or quantifying Human Immunodeficiency Virus-2 (HIV-2) nucleic acids in a biological sample taken from the individual as defined in  claim 4 ; 
 (b) determining therefrom whether the individual is infected by HIV-2 or the HIV-2 viral load of the individual. 
 
     
     
       12. An in vitro method for diagnosing HIV-2 infection, or determining HIV-2 viral load, in an individual, comprising the steps of:
 (a) performing the method for detecting or quantifying Human Immunodeficiency Virus-2 (HIV-2) nucleic acids in a biological sample taken from the individual as defined in  claim 5 ; 
 (b) determining therefrom whether the individual is infected by HIV-2 or the HIV-2 viral load of the individual.