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Plans for International Standard for HIV-2 RNA and 2 nd Second International Reference Panel for HIV-1 RNA Genotypes. Harvey Holmes, Clare Morris and Neil Berry Division of Retrovirology NIBSC, UK. (In collaboration with Indira Hewlett, CBER/FDA).

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Plans for International Standard for HIV-2 RNA and 2ndSecond International Reference Panel for HIV-1 RNA Genotypes

Harvey Holmes, Clare Morris and Neil Berry

Division of Retrovirology

NIBSC, UK

(In collaboration with Indira Hewlett, CBER/FDA)

current hiv reference reagents available from nibsc
Current HIV Reference Reagents Available from NIBSC
  • 2nd International Standard for HIV-1 RNA.

(code 97/650)

    • Unitage 5.56 IU Log 10/ml
  • International Reference Panel for HIV-1 RNA Genotypes (code 01/466) established in 2003
    • No unitage assigned
  • Working reagents for NAT
    • Working Reagent 1 – medium copy number (3.56 log10 IU/ml)
    • Working Reagent 2 – high copy number (4.56 log10 IU/ml)
    • Working Reagent 3 – low copy number (2.56 log10 IU/ml)
  • Working Reagents and Proficiency Samples can also be made at NIBSC under contract to a pre-defined concentration and using various HIV-1 strains and genotypes
international standard for hiv 2 rna
International Standard for HIV-2 RNA
  • Rationale
    • Global HIV pandemic due to HIV-1
    • Most NAT assays aimed at HIV-1 detection and quantification
    • HIV-2 mainly found in West Africa and European countries with close links – eg Portugal, France, Belgium.
    • Assays capable of detecting HIV-2 RNA or both HIV-1/HIV-2 are either available (eg Roche Cobas Taqscreen MPX) or in development
    • International Standard for the HIV-2 RNA component would be valuable once assays in wider use
  • Anticipated need/usage
    • Currently limited demand
    • Once commercial assays that can detect HIV-2 more available, demand may be greater
  • WHO Status
    • New project – endorsed by ECBS in October 2006
    • Discussed at meeting of WHO Collaborating Centres in Jan 07
key scientific issues
Key Scientific issues
  • Few commercial assays available that can detect/quantitate HIV-2 RNA
  • At NIBSC, in house real-time PCR assay based on LTR sequence used for HIV-2 quantification
  • Source/Type of Material
    • To be based on HIV-2 grown in culture and diluted in negative plasma
    • Currently assessing representatives of HIV-2 subtype A and subtype B
    • Suitable strain(s) to be selected and stock laid down
  • Virus stock to be characterised and sequence confirmed
  • Heat inactivation to be evaluated
  • Batch to be freeze-dried
  • International collaborative study to be organised
1 st international reference panel for hiv 1 rna genotypes code 01 466
1st International Reference Panel for HIV-1 RNA Genotypes (code 01/466)
  • Established by ECBS in 2003
  • 500 panels prepared and frozen at -80C
  • Contains representatives of HIV-1:
    • Subtypes A, B, C, D, AE, F, G, AGH
    • Group N and O
  • ~120 panels remain
  • Stability at -80C very good over 5 years
  • Proposal for 2nd IRP
second genotype panel for hiv 1 rna
Second Genotype Panel for HIV-1 RNA
  • Rationale
    • Many inter-subtype recombinant forms now circulating (CRF01 – CRF32)
    • Increasingly being encountered – important pandemic strains
    • Less common subtypes may be difficult to detect
    • Proposal to prepare extended panel containing:
      • Subtypes G, H, J and K, group N and O
      • Range of CRFs
      • Will give kit manufacturers and others access to rare and challenging strains of HIV to enable them to assess their ability to detect these viruses
  • Anticipated need/usage
    • Discussed at SoGAT working group meeting
    • Limited need
    • Provides a source of well characterised diverse CRFs and isolates
  • WHO Status
    • Project endorsed by ECBS October 2006
    • Discussed at meeting of WHO Collaborating Centres in Jan 07
second genotype panel for hiv 1 rna1
Second Genotype Panel for HIV-1 RNA
  • Source/Type of Material
    • Only relatively small number of characterised viruses belonging to uncommon subtypes or CRFs available as infectious virus
      • many only available as cloned DNA
    • Will source viruses from CFAR/NIBSC, CBER and NIH ARRRP and from scientists who have described them in the literature
    • Viruses to be grown in PBMC culture and seed stocks stored down
    • Viruses will be characterised including sequence confirmation
    • Viruses will be spiked into HIV-negative plasma
    • Heat inactivation and freeze-drying to be evaluated
    • International collaborative study to be organised
hiv field isolates reference strains and crfs currently available
Subtype G

Subtype H

Subtype J

Subtype K

Group N (to be sourced)

Group O

HIV-2 Subtype A

HIV-2 subtype B

CRF01_AE

CRF02_AG

CRF04_AGHKU (plasma only)

CRF07_BC

CRF11_A01GJ

CRF13_A01GJU

CRF14_BG

HIV field isolates, reference strains and CRFs currently available