Feasibility and efficacy of routine PCR screening of blood donations for hepatitis C virus, hepatitis B virus, and HIV-1 in a blood-bank setting

doi:10.1016/S0140-6736(98)06318-1

The Lancet

Volume 353, Issue 9150, 30 January 1999, Pages 359-363

https://doi.org/10.1016/S0140-6736(98)06318-1 Get rights and content

Summary

Background

Despite sensitive antibody-based blood-donor screening, a residual risk of transfusion-transmitted viral infections exists. Only direct monitoring by sensitive nucleic-acid tests would provide data accurately to measure the risk and to assess risk-reduction procedures. We investigated the feasibility and efficacy of routine screening of donors for hepatitis C virus (HCV), hepatitis B virus (HBV), and HIV-1 by PCR.

Methods

For PCR testing, individual donor plasma samples were pooled (96X100 μL) overnight by two automatic pipetting machines. Viruses were concentrated by centrifugation and nucleic acids were extracted. HCV PCR was done on the Cobas Amplicor system (Hoffmann-La Roche, Mannheim, Germany). HBV and HIV-1 sequences were amplified by single (non-nested) in-house PCRs and detected by agarose-gel electrophoresis. Detection limits were 1000–5000 genome equivalents/mL in the donor blood.

Findings

PCR testing was done in parallel to antibody screening with a maximum throughput of 3000 samples in 7–8 h. Positive samples were identified 1–2 days later. 111 of 373 423 donations (107 of 4500 pools) were PCR and antibody/antigen-confirmed positive. We found one HCV PCR-positive antibody-negative donation with normal alanine aminotransferase and one HCV PCR-positive donation with an elevated alanine aminotransferase (100 IU), which was negative in the AxSYM 2·0 and Matrix 1·0, but positive after control in the Abbott Prism test (Abbott GmbH, Wiesbaden, Germany).

Interpretation

PCR is a suitable and fast blood-donor screening procedure and contributes to a reduction in viral transmission by transfusion of blood components. In our selected donor population, the yield of detected contaminated donations from donors in the time window in which they are highly infectious but do not have any symptoms or detectable antigen and antibody concentrations (diagnostic window), confirms theoretical estimates.

Introduction

Virus safety of cellular blood components mainly relies on donor selection and donor screening for transfusionassociated viruses by antibody/antigen testing. The introduction of highly sensitive and specific secondgeneration and third-generation screening assays for hepatitis C virus (HCV) and HIV-1 antibodies and hepatitis B virus (HBV) antigens significantly reduced transmission risk.1, 2, 3, 4 However, transmission of these viruses by blood transfusion still occurs.1, 4 This is mostly due to the diagnostic window during which an acutely infected blood donor may harbour large amounts of highly infectious viruses without developing symptoms or detectable antigen and antibody concentrations.1, 3, 4, 5

Testing nucleic acids by highly sensitive amplification methods is a promising approach to shorten the preseroconversion window and reduce virus transmission. As calculated by Schreiber and colleagues,¹ PCR testing for HCV, HBV, and HIV-1 of blood donors would reduce this window by 59 days, 25 days, and 11 days, respectively. Virus-prevalence data for people who have donated blood many times (multiple-time donors) in the USA suggest that the residual risk of transmitting one of these viruses can be reduced by 72%, 42%, and 50%, respectively, with PCR. Based on these data and estimates of incidence of HCV, HBV, and HIV-1 infections in Germany6, 7 we decided to establish a PCRscreening procedure at our blood bank. The system should be suitable to test up to 3000 donations per day for HCV, HBV, and HIV-1.

We present the feasibility of PCR routine screening in a blood-bank setting and the results of testing 373 423 donations.

Section snippets

Routine donor screening by antigen/antibody ELISA

Routine donor screening for HCV, HBV, and HIV-1 was done with the Abbott Prism and Abbott AxSYM test systems (Abbott GmbH, Wiesbaden, Germany). Initially reactive samples were confirmed with the Abbott Matrix 1·0 and 2·0 (since January 1998) for HCV, Abbott HBsAg confirmatory test for HBV, and Sanofi Pasteur western blot for HIV-1.

Plasma-sample pooling

For PCR testing, 7 mL of blood was collected from each donor into an edetic-acid tube. Plasma was separated from cells within 12 h. Pooling was done overnight on two

Performance data

Using mini pools of 96 samples we had a maximum capacity of testing 3000 samples per day for HCV, HBV, and HIV-1 by PCR. Testing was completed in 7–8 h, enabling release of packed red cells and random platelet concentrates without delay. Identification of positive samples was achieved 1 or 2 days later and the retained negative-testing units of the pool were released.

Test results proved to be reliable and sensitive. The detection limits for HCV, HBV, and HIV-1 PCRs are shown in table 1. The

Discussion

We have established a sensitive and reliable routine HCV, HBV, and HIV-1 PCR for a blood-bank setting enabling testing of up to 3000 samples per day in 7–8 h.

Our efforts to establish a PCR for donor screening were mainly driven by the demands of producers of plasma products who wanted to use PCR-tested plasma from the beginning of 1997. Moreover, it became apparent that the Paul Ehrlich Institute, the governmental authority for the regulation of blood and blood products affairs of Germany,

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ArticlesFeasibility and efficacy of routine PCR screening of blood donations for hepatitis C virus, hepatitis B virus, and HIV-1 in a blood-bank setting

Summary

Background

Methods

Findings

Interpretation

Introduction

Section snippets

Routine donor screening by antigen/antibody ELISA

Plasma-sample pooling

Performance data

Discussion

Anal Biochem

The risk of transfusion-transmitted viral infections

N Engl J Med

Performance of three generations of anti-hepatitis C virus enzyme-linked immunosorbent assays in donors and patients

Transfusion

Estimated risk of transmission of the human immunodeficiency virus by screened blood in the United States

N Engl J Med

Food and drug administration conference on the feasibility of genetic technology to close the HIV window in donor screening

Transfusion

A long-term study of hepatitis C virus replication in non-A, non-B hepatitis

N Engl J Med

HIV, HCV and HBV in blood donors

Infusionsther Transfusionsmed

Prevalence and incidence of anti-HCV in 2·8 million blood donations in lower Saxony-HCV risk associated with transfusions [German]

PCR detection of hepatitis B virus

Articles
Feasibility and efficacy of routine PCR screening of blood donations for hepatitis C virus, hepatitis B virus, and HIV-1 in a blood-bank setting