Skip to main content
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems
  • Log in
  • My alerts
  • My Cart

Main menu

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • COVID-19 Special Collection
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About JCM
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems

User menu

  • Log in
  • My alerts
  • My Cart

Search

  • Advanced search
Journal of Clinical Microbiology
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • COVID-19 Special Collection
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About JCM
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
CASE REPORTS

Group O Human Immunodeficiency Virus Type 1 Infection That Escaped Detection in Two Immmunoassays

Said Zouhair, Sylvie Roussin-Bretagne, Alain Moreau, Sylvie Brunet, Syria Laperche, Michèle Maniez, Francis Barin, Martine Harzic
Said Zouhair
1Laboratoire de Virologie, Département de Biologie Médicale
3Service de Microbiologie, Hôpital Militaire d'Instruction Mohamed V, Rabat, Morocco
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Sylvie Roussin-Bretagne
2Service Hépato-Gastroentérologie, Centre Hospitalier de Versailles, Le Chesnay, France
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Alain Moreau
4Université François-Rabelais, Centre National de Référence du VIH, Tours, France
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Sylvie Brunet
4Université François-Rabelais, Centre National de Référence du VIH, Tours, France
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Syria Laperche
5Centre National de Transfusion Sanguine, Paris, France
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Michèle Maniez
6ETS de Lille, Lille, France
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Francis Barin
4Université François-Rabelais, Centre National de Référence du VIH, Tours, France
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Martine Harzic
1Laboratoire de Virologie, Département de Biologie Médicale
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: mharzic@ch-versailles.fr
DOI: 10.1128/JCM.44.2.662-665.2006
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

ABSTRACT

We report a case of human immunodeficiency virus type 1 group O infection not detected by two highly sensitive immunoassays. The sera were strongly reactive to the V3 peptide of group O but not to the gp41 immunodominant epitope. Gp41 was sequenced, confirming that this virus belonged to group O.

CASE REPORT

Screening for human immunodeficiency virus (HIV) antibodies was performed on a 42-year-old Cameroonian woman during her first pregnancy, 3 months after her arrival in France, in July 1994. At that time, serum was reactive in two second-generation enzyme-linked sorbent immunoassays (ELISAs), a competitive assay (Welcozyme HIV recombinant; Murex Diagnostics, Dartford, United Kingdom) and an indirect assay (Axsym HIV-1/2; Abbott Diagnostics, North Chicago, Ill.), respectively (Table 1). Seropositivity was confirmed by Western blotting (New LAV Blot 1; Bio-Rad, Marnes-la-Coquette, France) showing strong reactivity to the Gag, Pol, and Env proteins except to gp120. Her CD4+ T-cell count was 317/mm3, and p24 antigenemia was undetectable (Murex HIV antigen monoclonal antibody). The patient was asymptomatic. Antiretroviral treatment was initiated in 1995 because her CD4 cell count was low, close to 300/mm3. She was treated first with zidovudine and then with a zidovudine-zalcitabine combination, which allowed an increase of 100 CD4+ T cells/mm3. However, she developed pancreatitis and myositis in November 1997, which necessitated discontinuation of the antiretroviral regimen.

View this table:
  • View inline
  • View popup
TABLE 1.

Reactivities of sequential samples from the patient in different ELISAs

As a moderate decrease of her CD4 cell count was observed, a highly active antiretroviral treatment was instituted in 1998 (zidovudine, lamivudine, and indinavir), after which the CD4 cell count increased to 700/mm3. Her plasma virus load was undetectable in all the samples tested using either HIV RNA 3.0 (HIV-1 bDNA) (Bayer Diagnostics, Berkeley, CA) or Roche Amplicor HIV-1 Monitor v1.5 (HIV-1 Monitor) (Roche Molecular Systems, Branchburg, N.J.). Highly active antiretroviral treatment was discontinued in September 2001 because of treatment intolerance and a CD4 cell count close to 700/mm3.

In 2003, after delivery of her second child, a complete analysis of virological markers (detection of HIV antibodies, lymphocyte culture, and plasma HIV RNA quantification) was performed on samples from the mother and infant. Surprisingly, ELISAs for HIV antibodies gave negative results for both the mother and the newborn using a third-generation (HIV-1/2 Access; Bio-Rad, France) and a fourth-generation (Vidas Duo; Biomérieux, Marcy l'Étoile, France) assay. Previously collected sera that were retested using these assays were also found to be negative for HIV-1 antibodies (Table 1). Lymphocyte cultures were negative both for the mother's and newborn's samples, as was HIV RNA detection for the infant. The maternal viral load at delivery was 5,568 copies/ml using the LCx HIV RNA assay (Abbott Diagnostics) but was undetectable by both the HIV-bDNA and Monitor assays. The discordant data obtained in both serological assays and molecular assays strongly suggested infection by an unusual HIV-1 variant.

A peptide-based serotyping assay was therefore performed with two serum samples collected from the patient. This assay is an ELISA based on synthetic V3 loop and gp41 peptides to discriminate between HIV-1 group M, HIV-1 group O, and HIV-2 infections (1, 8). The serum samples were strongly reactive to the V3 peptide of HIV-1 group O, suggesting infection by a group O variant (Table 2). However, and quite unusually in our experience, antibodies to the gp41 immunodominant epitope (IDE) were not detected. A group-specific reverse transcriptase PCR able to discriminate between HIV-1 groups M and O was performed using primers located in the transmembrane region of the env gene and covering the IDE (9). A DNA fragment was obtained only with group O primers. The amplified segment was sequenced (392 nucleotides), and identification of the strain was done using the neighbor-joining method. The sequence was compared with 50 reference sequences corresponding to the nine subtypes and major circulating recombinant forms (CRF01-AE and CRF02-AG) of HIV-1 group M and to different strains of HIV-1 group O, available from the HIV sequence database (http://hiv-web.lanl.gov ). Distances were calculated with the Kimura two-parameter method, as implemented in the MEGA program. Bootstrap analysis with 100 simulations was used to test the reliability of branching. The strain clearly belonged to group O (Fig. 1).

FIG. 1.
  • Open in new tab
  • Download powerpoint
FIG. 1.

Phylogenetic analysis of gp41 of the case strain. Values at nodes indicate the percentages of bootstraps in which the cluster to the right was found. The tree was rooted using the SIVCPZ CAM5 sequence as an outgroup.

View this table:
  • View inline
  • View popup
TABLE 2.

Sequences of the immunodominant region (IDE) of gp41

Failure of the sera from this patient to bind to the group O IDE consensus peptide might be attributed to an unusual dipeptide motif (TR) located within the five-amino-acid loop of the IDE of the infecting variant (Table 2). Indeed, this TR motif was never found in any of the 64 available gp41 sequences of HIV-1 group O. Based on this observation, we investigated whether this divergent sequence was responsible for the lack of reactivity of the patient's sera to the group O IDE consensus peptide by preparing a synthetic peptide overlapping the IDE of the case's strain (see the sequence in Table 2). Serum samples from the case were tested in parallel by ELISA for this peptide, consensus peptide of IDE group O, and consensus peptide of IDE group M (1 μg/ml) based on a procedure described elsewhere (1). As shown in Table 3, the two serum samples from this patient bound the homologous IDE peptide, albeit with a relatively weak signal.

View this table:
  • View inline
  • View popup
TABLE 3.

Binding properties of the case's sera or control sera for various IDE and V3 peptides

Sequential sera collected between 1994 and 2004 and preserved at −20°C were tested, when available in sufficient amount, with various immunoassays (Table 1). The results were positive with most second- and third-generation assays, except HIV-1/2 Access, and negative with two of the four fourth-generation tested assays (Genscreen plus; Bio-Rad and Vidas Duo; bioMérieux).

Over the past 20 years, considerable progress has been made in the abilities of immunoassays to detect antibodies to groups M, N, and O of HIV-1 as well as antibodies to HIV-2 (5). Nevertheless, failure to detect HIV antibodies due to HIV genetic diversity has been regularly reported (3). This case is remarkable because one third-generation and one fourth-generation ELISA each failed to detect HIV infection, whereas most second- and third-generation assays did detect HIV infection. Most of the third-generation assays include a recombinant p24 protein, which is an antigen conserved between HIV groups. Third- and fourth-generation assays use peptides or recombinant proteins, including the gp41 immunodominant epitope from HIV-1 groups M and O and HIV-2 (2). The unique IDE sequence of the group O variant described herein might explain the failure by several immunoassays. This hypothesis is supported by the fact that antibodies from the patient bound its homologous sequence, although weakly, whereas they did not bind a group O consensus sequence. Since the Access assay that includes a p24 antigen was negative, our results indicate that even addition of this conserved antigen does not guarantee 100% sensitivity in detecting HIV variants.

Most HIV-1 RNA quantification assays detect only HIV-1 group M, except the LCx assay, which detects group O; so discordant results between different assays can suggest infection by a variant HIV virus.

Although the prevalence of HIV-1 group O is low even in areas of endemicity, such as Cameroon (6), this case report highlights the impact of its high genetic diversity (4, 7) on efficacy of diagnostic tools and illustrates how the genetic diversity of HIVs in general challenges the diagnostics field.

Nucleotide sequence accession number.The sequence of the 392-nucleotide DNA fragment obtained with group O primers in this study has been submitted to GenBank and assigned accession no. DQ222454.

FOOTNOTES

    • Received 5 July 2005.
    • Returned for modification 22 August 2005.
    • Accepted 18 November 2005.
  • Copyright © 2006 American Society for Microbiology

REFERENCES

  1. 1.↵
    Brand, D., A. Beby-Defaux, M. Mace, S. Brunet, A. Moreau, C. Godet, X. Jais, F. Cazein, C. Semaille, and F. Barin. 2004. First identification of HIV-1 groups M and O dual infections in Europe. AIDS3:2425-2428.
    OpenUrl
  2. 2.↵
    Dorn, J., S. Masciotra, C. Yang, R. Downing, B. Biryahwaho, T. D. Mastro, J. Nkengasong, D. Pieniazek, M. A. Rayfield, D. J. Hu, and R. B. Lal. 2000. Analysis of genetic variability within the immunodominant epitopes of envelope gp41 from human immunodeficiency virus type 1 (HIV-1) group M and its impact on HIV-1 antibody detection. J. Clin. Microbiol.38:773-780.
    OpenUrlAbstract/FREE Full Text
  3. 3.↵
    Gaudy, C., A. Moreau, S. Brunet, J. M. Descamps, P. Deleplanque, D. Brand, and F. Barin. 2004. Subtype B human immunodeficiency virus (HIV) type 1 mutant that escapes detection in a fourth-generation immunoassay for HIV infection. J. Clin. Microbiol.42:2847-2849.
    OpenUrlAbstract/FREE Full Text
  4. 4.↵
    Loussert-Ajaka, I., M. L. Chaix, B. Korber, F. Letourneur, E. Gomas, E. Allen, T. D. Ly, F. Brun-Vezinet, F. Simon, and S. Saragosti. 1995. Variability of human immunodeficiency virus type 1 group O strains isolated from Cameroonian patients living in France. J. Virol.69:5640-5649.
    OpenUrlAbstract/FREE Full Text
  5. 5.↵
    Ly, T. D., S. Laperche, and A. M. Courouce. 2001. Early detection of human immunodeficiency virus infection using third- and fourth-generation screening assays. Eur. J. Clin. Microbiol. Infect. Dis.20:104-110.
    OpenUrlCrossRefPubMed
  6. 6.↵
    Peeters, M., A. Gueye, S. Mboup, F. Bibollet-Ruche, E. Ekaza, C. Mulanga, R. Ouedrago, R. Gandji, P. Mpele, G. Dibanga, B. Koumare, M. Saidou, E. Esu-Williams, J. P. Lombart, W. Badombena, N. Luo, M. Vanden Haesevelde, and E. Delaporte. 1997. Geographical distribution of HIV-1 group O viruses in Africa. AIDS11:493-498.
    OpenUrlCrossRefPubMedWeb of Science
  7. 7.↵
    Roques, P., D. L. Robertson, S. Souquiere, F. Damond, A. Ayouba, I. Farfara, C. Depienne, E. Nerrienet, D. Dormont, F. Brun-Vezinet, F. Simon, and P. Mauclere. 2002. Phylogenetic analysis of 49 newly derived HIV-1 group O strains: high viral diversity but no group M-like subtype structure. Virology302:259-273.
    OpenUrlCrossRefPubMed
  8. 8.↵
    Simon, F., S. Souquiere, F. Damond, A. Kfutwah, M. Makuwa, E. Leroy, P. Rouquet, J. L. Berthier, J. Rigoulet, A. Lecu, P. T. Telfer, I. Pandrea, J. C. Plantier, F. Barre-Sinoussi, P. Roques, M. C. Muller-Trutwin, and C. Apetrei. 2001. Synthetic peptide strategy for the detection of and discrimination among highly divergent primate lentiviruses. AIDS Res. Hum. Retrovir.17:937-952.
    OpenUrlCrossRefPubMedWeb of Science
  9. 9.↵
    Yang, C., D. Piezanek, S. M. Owen, C. Fridlund, J. Nkengasong, T. D. Mastro, M. A. Rayfield, R. Downing, B. Biryawaho, A. Tanuri, L. Zekeng, G. van derGroen, F. Gao, and R. Lal. 1999. Detection of phylogenetically diverse human immunodeficiency virus type 1 groups M and O from plasma by using highly sensitive and specific generic primers. J. Clin. Microbiol.37:2581-2586.
    OpenUrlAbstract/FREE Full Text
View Abstract
PreviousNext
Back to top
Download PDF
Citation Tools
Group O Human Immunodeficiency Virus Type 1 Infection That Escaped Detection in Two Immmunoassays
Said Zouhair, Sylvie Roussin-Bretagne, Alain Moreau, Sylvie Brunet, Syria Laperche, Michèle Maniez, Francis Barin, Martine Harzic
Journal of Clinical Microbiology Feb 2006, 44 (2) 662-665; DOI: 10.1128/JCM.44.2.662-665.2006

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Print

Alerts
Sign In to Email Alerts with your Email Address
Email

Thank you for sharing this Journal of Clinical Microbiology article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Group O Human Immunodeficiency Virus Type 1 Infection That Escaped Detection in Two Immmunoassays
(Your Name) has forwarded a page to you from Journal of Clinical Microbiology
(Your Name) thought you would be interested in this article in Journal of Clinical Microbiology.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Group O Human Immunodeficiency Virus Type 1 Infection That Escaped Detection in Two Immmunoassays
Said Zouhair, Sylvie Roussin-Bretagne, Alain Moreau, Sylvie Brunet, Syria Laperche, Michèle Maniez, Francis Barin, Martine Harzic
Journal of Clinical Microbiology Feb 2006, 44 (2) 662-665; DOI: 10.1128/JCM.44.2.662-665.2006
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • CASE REPORT
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

KEYWORDS

HIV Antibodies
HIV Infections
HIV-1
Immunodominant Epitopes
Pregnancy Complications, Infectious

Related Articles

Cited By...

About

  • About JCM
  • Editor in Chief
  • Board of Editors
  • Editor Conflicts of Interest
  • For Reviewers
  • For the Media
  • For Librarians
  • For Advertisers
  • Alerts
  • RSS
  • FAQ
  • Permissions
  • Journal Announcements

Authors

  • ASM Author Center
  • Submit a Manuscript
  • Article Types
  • Resources for Clinical Microbiologists
  • Ethics
  • Contact Us

Follow #JClinMicro

@ASMicrobiology

       

ASM Journals

ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology.

About ASM | Contact Us | Press Room

 

ASM is a member of

Scientific Society Publisher Alliance

 

American Society for Microbiology
1752 N St. NW
Washington, DC 20036
Phone: (202) 737-3600

 

Copyright © 2021 American Society for Microbiology | Privacy Policy | Website feedback

Print ISSN: 0095-1137; Online ISSN: 1098-660X