Reference panel of cloned HIV-2 plasmid DNA for nucleic acid assay development, evaluation, and quality monitoring

J Clin Virol. 2014 Oct;61(2):293-7. doi: 10.1016/j.jcv.2014.06.028. Epub 2014 Jul 7.

Abstract

Background: Currently, no FDA-approved HIV-2 nucleic acid assay is commercially available in the United States, although several laboratories have developed in-house assays to confirm HIV-2 infections. A major limitation in the development of novel HIV-2 diagnostic assays is the lack of reference materials that can be used to evaluate, optimize, and monitor assay performance.

Study design: Eleven viral stocks of HIV-2 isolates from various West African countries, including the Ivory Coast, Senegal, and Guinea-Bissau, were used to clone the entire LTR and pol regions from each virus.

Results: We successfully cloned, sequenced, and group classified 22 HIV-2 DNA plasmids including 11 full length LTR (∼849 bp) and 11 pol (∼2995 bp) sequences. There were eight HIV-2 group A and three group B in both the LTR and pol regions.

Conclusions: This reference panel provides a robust, quantifiable, renewable, and non-infectious set of reagents that can be used for the development and evaluation of new HIV-2 molecular diagnostic assays and quality assurance and quality control reagents for use in the clinical laboratories.

Keywords: Clone; HIV-2; HIV-2 group; NAT; Plasmid; Real-time PCR.

MeSH terms

  • DNA, Viral / genetics
  • HIV Infections / diagnosis*
  • HIV Long Terminal Repeat / genetics
  • HIV-2 / genetics
  • HIV-2 / isolation & purification*
  • Humans
  • Molecular Diagnostic Techniques / methods
  • Molecular Diagnostic Techniques / standards*
  • Plasmids
  • Real-Time Polymerase Chain Reaction / methods
  • Real-Time Polymerase Chain Reaction / standards*
  • Reference Standards*
  • Vereinigte Staaten
  • pol Gene Products, Human Immunodeficiency Virus / genetics

Substances

  • DNA, Viral
  • pol Gene Products, Human Immunodeficiency Virus