Blood screening nucleic acid amplification tests for human immunodeficiency virus Type 1 may require two different amplification targets
- 2 August 2011
- journal article
- case report
- Published by Wiley in Transfusion
- Vol. 52 (2), 431-439
- https://doi.org/10.1111/j.1537-2995.2011.03281.x
Abstract
Five cases of human immunodeficiency virus Type 1 (HIV-1) RNA-positive blood donations are described that escaped detection by three different CE-marked nucleic acid amplification technique (NAT) screening assays. These events were associated with two HIV-1 transmissions to recipients of blood components. The implicated NAT assays are monotarget assays and amplify in different viral genome regions (group-specific antigen or long terminal repeat). Investigations into the cause of the false-negative test results were initiated. Plasma specimens of the five NAT false-negative cases were comparatively investigated in 12 CE-marked HIV-1 NAT systems of differing design. The relative amplification efficiency for the HIV-1 variant was determined for each assay. Sequencing of the variants in the region targeted by each false-negative NAT assay allowed comparison with the respective primers and probes. Some of the NAT assays designed in a similar way to false-negative monotarget NATs also revealed deficiencies in detecting the viral variants. In each case sequencing of the assay target region in the variants demonstrated mismatches with primers and probes used by the assays. Some dual-target assays showed decreased amplification efficiency, but not false-negative results. HIV is characterized by its rapid evolution of new viral variants. The evolution of new sequences is unpredictable; NAT screening assays with a single target region appear to be more vulnerable to sequence variations than dual-target assays. Based on this experience with false-negative tests results by monotarget NAT assays, the Paul-Ehrlich-Institut is considering requesting dual-target NAT assays for HIV-1 blood donation screening in Germany.Keywords
This publication has 13 references indexed in Scilit:
- Infektionsepidemiologische Daten von Blutspendern in Deutschland 2007Bundesgesundheitsblatt - Gesundheitsforschung - Gesundheitsschutz, 2010
- Performance Evaluation of the New Roche Cobas AmpliPrep/Cobas TaqMan HIV-1 Test Version 2.0 for Quantification of Human Immunodeficiency Virus Type 1 RNAJournal of Clinical Microbiology, 2010
- The Effect of Primer-Template Mismatches on the Detection and Quantification of Nucleic Acids Using the 5′ Nuclease AssayThe Journal of Molecular Diagnostics, 2010
- Performance of the New Roche Cobas AmpliPrep-Cobas TaqMan Version 2.0 Human Immunodeficiency Virus Type 1 AssayJournal of Clinical Microbiology, 2009
- Experience of mandatory nucleic acid test (NAT) screening across all blood organizations in Germany: NAT yield versus breakthrough transmissionsTransfusion, 2009
- First transmission of human immunodeficiency virus Type 1 by a cellular blood product after mandatory nucleic acid screening in GermanyTransfusion, 2009
- Ultrasensitive Monitoring of HIV-1 Viral Load by a Low-Cost Real-Time Reverse Transcription-PCR Assay with Internal Control for the 5′ Long Terminal Repeat DomainClinical Chemistry, 2006
- An international collaborative study to establish the 1st international standard for HIV-1 RNA for use in nucleic acid-based techniquesJournal of Virological Methods, 2001
- Validation of HCV-NAT Assays and Experience with NAT Application for Blood Screening in GermanyBiologicals, 1999
- The effects of internal primer-template mismatches on RT-PCR: HIV-1 model studiesNucleic Acids Research, 1997