In vitro method for the detection and quantification of HIV-2
View Patent ↗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.
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.