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Journal of Clinical Microbiology, March 2007, p. 715-720, Vol. 45, No. 3
0095-1137/07/$08.00+0     doi:10.1128/JCM.01264-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Comparative Study of Kaposi's Sarcoma-Associated Herpesvirus Serological Assays Using Clinically and Serologically Defined Reference Standards and Latent Class Analysis{triangledown}

Maria Claudia Nascimento,1,2* Vanda Akico de Souza,2 Laura Masami Sumita,2 Wilton Freire,2 Fernando Munoz,3 Joseph Kim,4 Claudio S. Pannuti,2 and Philippe Mayaud1

Clinical Research Unit, Department of Infectious & Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT, United Kingdom,1 Laboratório de Virologia do Instituto de Medicina Tropical and Departamento de Moléstias Infecciosas e Parasitárias da Faculdade de Medicina, Universidade de São Paulo, Av. Dr. Eneas de Carvalho Aguiar 470, Sao Paulo, Brazil,2 The Cancer Research UK Viral Oncology Group, Wolfson Institute for Biomedical Research, University College of London Medical School, Cruciform Bldg., Gower Street, London WC1E 6BT, United Kingdom,3 Medical Statistics Unit, Department of Epidemiology and Population Health, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT, United Kingdom4

Received 20 June 2006/ Returned for modification 28 November 2006/ Accepted 14 December 2006

Accurate determination of infection with Kaposi's sarcoma-associated herpesvirus (KSHV) has been hindered by the lack of a "gold standard" for comparison of serological assays used to estimate KSHV prevalence in serosurveys conducted in different settings. We have evaluated the performance of five in-house (developed at University College London [UCL], United Kingdom, and at the virology laboratory of the Instituto de Medicine Tropical [IMT] in Sao Paulo, Brazil) and two commercial (ABI and DIAVIR) serological assays to detect antibodies to latency-associated nuclear antigen (LANA) and to lytic KSHV antigens. We used a variety of serum samples assembled to represent populations likely to be at high, intermediate, and low risk of KSHV infection in Brazil. Composite reference standard panels were prepared based on clinical and serological parameters, against which assay performances were assessed using conventional Bayesian statistics and latent class analysis (LCA). Against the clinical reference standard, in-house immunofluorescence assays to detect anti-LANA antibodies (IFA-LANA) produced at UCL and IMT had similar performances, with sensitivities of 61% (95% confidence interval [CI], 48% to 74%) and 72% (95% CI, 58% to 83%) and specificities of 99% (95% CI, 94% to 100%) and 100% (95% CI, 96% to 100%), respectively, and only the IMT IFA-LANA was included in LCA, together with the IMT IFA-lytic and four enzyme-linked immunosorbent assays (ELISAs). The LCA indicated that the IMT whole-virus ELISA performed best (sensitivity, 87% [95% CI, 81% to 91%]; and specificity, 100% [95% CI, 98% to 100%]), confirming the results obtained with the conventional statistical approach. Commercially available ELISA-based tests yielded the lowest specificities using a spectrum of serum samples. The evaluation of KSHV serological assays is warranted before planning serosurveys in various settings.

* Corresponding author. Present address: International Agency for Research on Cancer, 150 Cours Albert Thomas, 69372 Lyon Cedex 08, France. Phone: 33 472738179. Fax: 33 472738345. E-mail: nascimentoc{at}iarc.fr.

{triangledown} Published ahead of print on 20 December 2006.

Journal of Clinical Microbiology, March 2007, p. 715-720, Vol. 45, No. 3
0095-1137/07/$08.00+0     doi:10.1128/JCM.01264-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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