RESUMO
BACKGROUND: The incidence of hepatitis E virus (HEV) infection in the Netherlands is high. Blood donors are not routinely screened for HEV infection, but since January 2013, donations used for the production of solvent/detergent (S/D)-treated plasma have been screened for HEV RNA. STUDY DESIGN AND METHODS: Donations were screened for HEV RNA in pools of 96 and 192 donations. In addition, all donations made between 60 days before and after each HEV RNA-positive donation were tested individually for HEV RNA and anti-HEV immunoglobulin G. RESULTS: The screening of 59,474 donations between January 2013 and December 2014 resulted in identification of 45 HEV RNA-positive donations (0.076%) from 41 donors. HEV RNA loads ranged from 80 to 2.3 × 10(6) IU/mL. The number of positive donations increased significantly over time (p = 0.03). Thirty-three of 90 donations made up to 60 days before or after HEV RNA-positive donations were positive when tested individually, while they had not been detected in the pool screening. The mean duration of HEV viremia in the healthy blood donor is estimated to be 68 days. CONCLUSION: The incidence of HEV infection in the Netherlands is high and increased during the study period. In 2013 and 2014, HEV RNA was detected in 1 per 762 donations intended for production of S/D plasma.
Assuntos
Doadores de Sangue/estatística & dados numéricos , Vírus da Hepatite E/fisiologia , Hepatite E/epidemiologia , Hepatite E/patologia , Feminino , Vírus da Hepatite E/classificação , Vírus da Hepatite E/genética , Humanos , Incidência , Masculino , Países Baixos/epidemiologia , Filogenia , RNA Viral/genética , Estudos SoroepidemiológicosRESUMO
BACKGROUND: To meet European guidelines for plasma for fractionation, plasma fractionators have implemented parvovirus B19 (B19V) and hepatitis A virus (HAV) nucleic acid test (NAT) screening on test pools. In this study we evaluate recently developed in-house NAT assays for B19V DNA and HAV RNA. The B19V NAT was designed to target two different regions of the B19V genome. STUDY DESIGN AND METHODS: The B19V DNA and HAV RNA tests were validated according to commonly used guidelines. The performance of the B19V and HAV assays was evaluated during routine screening of more than 2 × 10(6) donations. RESULTS: The 95% lower limit of detection (LLD) of the HAV NAT was 1.34 IU/mL. The 95% LLD for B19V was 39.1 IU/mL for the NS1 region and 76.9 IU/mL for the VP2 region. The B19V test showed good accuracy, precision, robustness, and no cross-contamination was observed. Both assays detected B19V Genotypes 1 to 3 and HAV Genotypes I to III. During routine screening 103 donations showed B19V DNA loads of more than 1.25 × 10(6) IU/mL and one donation was reactive in the HAV NAT. CONCLUSION: The dual-target B19V polymerase chain reaction (PCR) showed good accuracy (<0.1 log IU/mL) at the crucial concentration of 10 IU/µL for the NS1 and the VP2 region of the B19V genome and detected all known genotypes with similar sensitivity for each genotype. In addition, the dual target format reduces the chance that molecular variants of B19V are wrongly quantified. The HAV RNA assay showed high sensitivity for Genotypes I to III. Both new PCR assays have been successfully introduced for plasma screening in test pools of 480 or 96 donations.
Assuntos
DNA Viral/sangue , Seleção do Doador/métodos , Vírus da Hepatite A , Parvovirus B19 Humano , RNA Viral/sangue , Reação em Cadeia da Polimerase em Tempo Real/métodos , Humanos , Guias de Prática Clínica como Assunto , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Routine HCV NAT minipool screening (48 donations) of all blood donations was implemented in July 1999 and was combined with HIV NAT in November 2000. This report describes the validation of the NAT methods and the results of quality control testing. STUDY DESIGN AND METHODS: Nucleic acid was extracted from 2-mL plasma samples by using an automated silica-based extraction method (NucliSens Extractor, Organon Teknika). Eluates were tested with RT-PCR (AmpliScreen HIV-1 version 1.5 and AmpliScreen HCV version 2.0 test, Roche Diagnostic Systems). HIV-1 and HCV RNA reference panels and run controls (PeliCheck and PeliSpy, respectively, Sanquin-CLB) and human plasma minipools were used for NAT validation. RESULTS: The 95-percent detection limit (and 95% CI) for HIV-1 RNA genotype B, HIV-1 RNA genotype E, and HCV RNA genotype 1 was 32 (19-76), 30 (17-72), and 21 (13-44) genome equivalents (geq) per mL, respectively. During initial validation, 2332 samples for HIV-1 RNA and 2644 samples for HCV RNA were analyzed, with 13 (0.56%) and 12 (0.45%) invalid test results, respectively. Thereafter, over 19,600 samples (minipools and run controls) were analyzed during the first 11 months of routine screening. Invalid test results for HIV-1 RNA and HCV RNA were found in 1.1 and 1.07 percent of the samples tested, respectively. HIV-1 RNA minipool testing resulted in 27 (0.16%) initial false-positive results and 3 (0.02%) confirmed positive results. HCV RNA minipool testing resulted in four (0.02%) initial false-positive results and five (0.02%) confirmed positive results. CONCLUSION: Routine HIV and HCV NAT minipool screening using the NucliSens Extractor, AmpliScreen HIV-1 version 1.5, and AmpliScreen HCV version 2.0 meets the sensitivity criteria set by the regulatory bodies and provides sufficient specificity and robustness for timely release of blood donations.
