Please cite this article in press as: Okonji, J.A., et al., Comparison of HIV-1 detection in plasma specimens and dried blood spots using the Roche COBAS Ampliscreen HIV-1 test in Kisumu, Kenya. J. Virol. Methods (2011), doi:10.1016/j.jviromet.2011.07.001 ARTICLE IN PRESS G Model VIRMET-11602; No. of Pages 5 Journal of Virological Methods xxx (2011) xxx–xxx Contents lists available at ScienceDirect Journal of Virological Methods jou rn al h om epage: www.elsevier.com/locate/jviromet Comparison of HIV-1 detection in plasma specimens and dried blood spots using the Roche COBAS Ampliscreen HIV-1 test in Kisumu, Kenya  Jully A. Okonji a , Sridhar V. Basavaraju b,∗ , Jane Mwangi c , Ray W. Shiraishi b , Matthew Odera d , Kenneth Ouma a , John P. Pitman b , Lawrence H. Marum b , Chin-Yih Ou b , Clement Zeh c a Centre for Biomedical Research and Technology, Kenya Medical Research Institute (KEMRI), Off Kisumu-Busia Road, P.O. Box 1578-40100, Kisumu, Kenya b U.S. Centers for Disease Control and Prevention, Center for Global Health, Division of Global HIV/AIDS, 1600 Clifton Road, NE MS E04, Atlanta, GA, USA c U.S. Centers for Disease Control and Prevention, Off Kisumu-Busia Road, P.O. Box 1578-401000, Kisumu, Kenya d Kenya National Blood Transfusion Service, Kisumu Regional Blood Transfusion Center, P.O. Box 9082, Kisumu, Kenya Article history: Received 4 April 2011 Received in revised form 30 June 2011 Accepted 4 July 2011 Available online xxx Keywords: HIV-1 Blood donors Nucleic acid testing Dried blood spots a b s t r a c t The World Health Organization recommends screening donor blood for HIV in centralized laboratories. This recommendation contributes to quality, but presents specimen transport challenges for resource- limited settings which may be relieved by using dried blood spots (DBS). In sub-Saharan Africa, most countries screen donor blood with serologic assays only. Interest in window period reduction has led blood services to consider adding HIV nucleic acid testing (NAT). The U.S. Food and Drug Administra- tion (FDA) mandates that HIV-1 NAT blood screening assays have a 95% detection limit at or below 100 copies/ml and 5000 copies/ml for pooled and individual donations, respectively. The Roche COBAS Ampliscreen HIV-1 test, version 1.5, used for screening whole blood or components for transfusion, has not been tested with DBS. We compared COBAS Ampliscreen HIV-1 RNA detection limits in DBS and plasma. An AIDS Clinical Trials Group, Viral Quality Assurance laboratory HIV-1 standard with a known viral load was used to create paired plasma and DBS standard nine member dilution series. Each was tested in 24 replicates with the COBAS Ampliscreen. A probit analysis was conducted to calculate 95% detection limits for plasma and DBS, which were 23.8 copies/ml (95% CI 15.1–51.0) for plasma and 106.7 copies/ml (95% CI 73.8–207.9) for DBS. The COBAS Ampliscreen detection threshold with DBS suggests acceptability for individual donations, but optimization may be required for pooled specimens. © 2011 Published by Elsevier B.V. 1. Introduction To reduce the risk of transfusion-transmitted HIV infections in resource-limited settings, WHO recommends universal blood screening for HIV in centralized laboratories (WHO, 2010). Cen- tralized donor blood screening, in one or few national laboratories, has been advocated by WHO for several reasons including optimiz- ing the cost effective use of resources and standardizing laboratory testing by eliminating variations between laboratory practices and testing results (WHO, 2002, 2010). Additionally, WHO recommen- dations advise a national system for the evaluation, selection and validation of all assays used for blood screening (WHO, 2010).  This work has been submitted as an abstract to the 2011 AABB Conference and Expo in San Diego, California. ∗ Corresponding author. Tel.: +1 404 639 8011; fax: +1 404 639 8105. E-mail addresses: jokonji@ke.cdc.gov (J.A. Okonji), etu7@cdc.gov (S.V. Basavaraju), jwmwangi@ke.cdc.gov (J. Mwangi), fnf3@cdc.gov (R.W. Shiraishi), kouma@ke.cdc.gov (K. Ouma), pitmanj@na.cdc.gov (J.P. Pitman), maruml@zm.cdc.gov (L.H. Marum), cho2@cn.cdc.gov (C.-Y. Ou), czeh@ke.cdc.gov (C. Zeh). The minimum evaluated sensitivity and specificity levels of blood screening assays should be at least 99.5% and quality-assured screening of donations with serologic assays should be instituted prior to implementing nucleic acid testing (NAT) of donor blood (WHO, 2010). At present in many sub-Saharan African countries, blood is often collected in remote locations via mobile collection drives and screened for HIV and other transfusion-transmitted infections in hospital-based or other decentralized laboratories. Algorithms for universal blood screening vary between high development index and resource-limited settings. In most high development index countries, donor blood is universally screened with nucleic acid testing (NAT) and with fourth generation p24 antigen and HIV-1 and -2 antibody immunoassays. Both NAT and fourth generation immunoassays have window periods during which acute HIV infections may not be detected. For NAT, the window period is approximately 6–15 days following infection (Busch et al., 2005; Corfec et al., 1999); the fourth generation assay window period is approximately 15–22 days (Busch et al., 2005; Ly et al., 2004). NAT is intended to find HIV infection prior to detectable levels of p24 antigen or HIV antibodies in blood, while the immunoassays are used to screen donor blood units from 0166-0934/$ – see front matter © 2011 Published by Elsevier B.V. doi:10.1016/j.jviromet.2011.07.001