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

a human immunodeficiency virus and in vitro method technology, applied in the field of in vitro method for the detection and quantification of human immunodeficiency virus (hiv) 2, can solve the problems of hampered clinical management of hiv-2 infection, difficult to accurately quantify group b viruses, and difficult to accurately diagnose hiv-2 infection

Active Publication Date: 2019-08-20
ASSISTANCE PUBLIQUE HOPITAUX DE PARIS +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The method provides improved sensitivity and accuracy, enabling detection and quantification of HIV-2 nucleic acids with a lower limit of quantification, effectively addressing the limitations of existing assays and facilitating better clinical management of HIV-2 infection.

Problems solved by technology

Nevertheless, HIV-2 infection eventually leads to AIDS.
Clinical management of HIV-2 infection is hampered by the lack of validated commercial RNA viral load assays.
Besides, the high genetic diversity of HIV-2, with 9 groups designated A to I, of which only groups A and B are epidemic, also represents an obstacle to accurate viral load quantification: previous studies have thus shown that group B viruses are particularly difficult to quantify with the current assays.
In addition, the current HIV-2 assays also suffer from low sensitivity and accuracy.

Method used

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

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Materials and Methods

HIV-2 RNA Assay According to the Invention

[0083]The test is based on a one-step duplex Taqman PCR approach targeting a conserved consensus region in the long terminal repeat (LTR) region and the Gag region.

[0084]The forward and reverse primers for the LTR region are 5′-TCTTTAAGCAAGCAAGCGTGG-3′ (SEQ ID NO: 1) and 5′-AGCAGGTAGAGCCTGGGTGTT-3′ (SEQ ID NO: 2), respectively (Rouet et al. (2004) J. Clin. Microbiol. 42:4147-53), with an internal probe (5′ FAM-CTTGGCCGGYRCTGGGCAGA-BHQ1 3′, SEQ ID NO: 3) modified to optimize efficiency for HIV-2 group B.

[0085]The forward and reverse primers for the Gag region are F3 5′-GCGCGAGAAACTCCGTCTTG-3′ (SEQ ID NO: 4) and R1 5′-TTCGCTGCCCACACAATATGTT-3′ (SEQ ID NO: 5), respectively (Damond et al. (2005) J. Clin. Microbiol. 43:4234-6), and the internal HIV-2 Taqman gag probe is S65GAG2 5′ FAM-TAGGTTACGGCCCGGCGGAAAGA-BHQ1 3′ (SEQ ID NO:6) (Eurogentec, Seraing, Belgium) (Damond et al. (2005) J. Clin. Microbiol. 43:4234-6).

[0086]

TABLE 1...

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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, andsequence SEQ ID NO: 2 or a sequence having at least 90% identity to SEQ ID NO: 2, andsequence SEQ ID NO: 4 or a sequence having at least 90% identity to SEQ ID NO: 4, andsequence 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, andsequence 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.

Description

FIELD OF THE INVENTION[0001]The present invention relates to an in vitro method for the detection and quantification of Human Immunodeficiency Virus (HIV) 2.TECHNICAL BACKGROUND[0002]HIV-2 is characterized by less efficient transmission through the sexual and vertical routes than HIV-1, and by a slower natural clinical course. Nevertheless, HIV-2 infection eventually leads to AIDS. HIV-2 infection must be distinguished from HIV-1 infection, as HIV-2 is naturally resistant to non-nucleoside reverse transcriptase inhibitors, T20, and some protease inhibitors, and as patient follow-up differs from that of HIV-1 infection.[0003]Compared to HIV-1, HIV-2 is characterized by lower viral replication. In the French ANRS cohort CO5 of HIV-2-infected patients (1009 patients in January 2014), 61% of untreated patients have plasma viral loads below 250 copies / mL (cp / mL). Likewise, in a British study, only 8% of patients with CD4>500 cells / mm3 and 62% of patients with CD4<300 cells / mm3 had ...

Claims

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Application Information

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Patent Type & AuthorityPatents(United States)
IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/703C12Q1/68C12Q2600/158C12Q2600/118
InventorROUZIOUX, CHRISTINEPLANTIER, JEAN-CHRISTOPHEAVETTAND-FENOEL, VÉRONIQUEDAMOND, FLORENCEGUEUDIN, MARIEDESCAMPS, DIANE
OwnerASSISTANCE PUBLIQUE HOPITAUX DE PARIS