Discrepant amplification results during the development of an assay leads to reclassification of two AIDS reagent repository HIV-2 isolates as HIV-1

Linda L. Jagodzinski, Ying Liu, Holly R. Hack, Catherine Kibirige, Sheila A. Peel, Mark M. Manak

Research output: Contribution to journalArticle

Abstract

The development and verification of HIV-2 assays depends in part on the availability of well-characterized samples, including those from reagent repositories. During the development of an HIV-2 RNA quantification assay, two HIV-2 viral isolates (CDC 301340 and CDC 301342) obtained from the NIAID AIDS Reagent and Reference Repository were not detected leading to an investigation. Two HIV-2 primers/probe sets of known performance in real-time viral RNA quantification assays, targeting different regions of the virus, also failed to generate RT-PCR products for these two isolates. These isolates were tested in the HIV-1 specific COBAS AmpliPrep/COBAS TaqMan HIV-1 Test v2.0 (Roche Molecular Diagnostics) and were quantified at high copy number. Other HIV-2 isolates tested were not amplified in the COBAS HIV-1 TaqMan assay. Furthermore, the discrepant isolates were highly reactive in an HIV-1 p24 antigen test while the other HIV-2 isolates showed very weak, if any, cross-reactivity with the HIV-1 p24 assay. Phylogenetic tree analysis of sequences from the protease-reverse transcriptase regions of the discrepant HIV-2 isolates mapped with HIV-1 Group M, Subtype CRF02-AG confirming these isolates were of HIV-1 origin and had been misclassified as HIV-2. The use of misclassified isolates in the verification of molecular and immunological assays can lead to misinterpretation of test results, misdirection of efforts into assay redesign and increased development costs. The results of this study were shared with the NIAID AIDS Reagent Program, leading to the reclassification of the two discrepant isolates as HIV-1.

Original languageEnglish (US)
Article numbere96554
JournalPLoS One
Volume9
Issue number5
DOIs
StatePublished - May 5 2014
Externally publishedYes

Fingerprint

Human immunodeficiency virus 2
HIV-2
taxonomic revisions
Human immunodeficiency virus 1
Amplification
HIV-1
Assays
Acquired Immunodeficiency Syndrome
assays
National Institute of Allergy and Infectious Diseases (U.S.)
Centers for Disease Control and Prevention (U.S.)
HIV Core Protein p24
RNA
RNA-Directed DNA Polymerase
Viral RNA
Molecular Pathology
RNA-directed DNA polymerase
Viruses
testing
immunoassays

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Discrepant amplification results during the development of an assay leads to reclassification of two AIDS reagent repository HIV-2 isolates as HIV-1. / Jagodzinski, Linda L.; Liu, Ying; Hack, Holly R.; Kibirige, Catherine; Peel, Sheila A.; Manak, Mark M.

In: PLoS One, Vol. 9, No. 5, e96554, 05.05.2014.

Research output: Contribution to journalArticle

Jagodzinski, Linda L. ; Liu, Ying ; Hack, Holly R. ; Kibirige, Catherine ; Peel, Sheila A. ; Manak, Mark M. / Discrepant amplification results during the development of an assay leads to reclassification of two AIDS reagent repository HIV-2 isolates as HIV-1. In: PLoS One. 2014 ; Vol. 9, No. 5.
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