Abstract
Background & AimsDried blood spots (DBS) on filter paper have been successfully used to diagnose and monitor several infectious diseases. The aim was to investigate the performance of DBS in hepatitis B virus (HBV) diagnosis using commercial tests in comparison to standard methods.MethodsPaired DBS and plasma samples were collected from 200 patients: 100 patients with HBsAg negative status and 100 patients with HBsAg positive status. In the latter patient, HBeAg reactivity was tested. Ten samples of anti-HBs were collected from people vaccinated against HBV. We also studied 50 patients with positive HBV DNA viral load in plasma and 10 HBV DNA negative patients. HBV genotypes and gene polymerase mutations were determined in 10 randomly selected HBV-infected patients. The DBS sample consisted of 50 µL of whole blood, i.e. a 12-mm paper card.ResultsThe sensitivity thresholds of HBsAg and anti-HBs antibody were 0.30±0.08 IU/mL and 18.11±6.05 IU/mL, respectively, for DBS with 98% sensitivity and 100% specificity. Sensitivity was 98% and specificity 100% for the detection of HBV DNA on a blotter, considering an HBV DNA threshold of 914.1±157.8 IU/ml. Ten patients had an HBeAg positive status in plasma, all were detected positive using DBS. HBV genotyping and mutation detection were successfully performed on DBS, with full concordance between the 10 paired DBS and plasma samples.ConclusionThis study shows DBS is a reliable alternative to plasma specimens for quantifying and detecting HBsAg, anti-HBs, HBeAg and genotyping. DBS may increase the opportunities for HBV testing and treatment follow-up in hard-to-reach individuals.
Highlights
About one third of the world’s population has serological evidence of past or present infection with hepatitis B virus (HBV) and 350 to 400 million people are chronic HBV surface antigen (HBsAg) carriers
The spectrum of disease and natural history of chronic HBV infection range from inactive carrier status to progressive chronic hepatitis B (CHB), which may evolve to cirrhosis and hepatocellular carcinoma (HCC) [1,2,3]
In HBV infection, dried blood spots (DBS) have been used for serology and detecting molecular biology markers such as HBV DNA, HBV core gene, anti-HBs, anti-HBc, HBsAg, hepatitis B e antigen (HBeAg) and for genotyping [15,16,17,18,19]
Summary
About one third of the world’s population has serological evidence of past or present infection with hepatitis B virus (HBV) and 350 to 400 million people are chronic HBV surface antigen (HBsAg) carriers. In HBV infection, DBS have been used for serology and detecting molecular biology markers such as HBV DNA, HBV core gene, anti-HBs, anti-HBc, HBsAg, hepatitis B e antigen (HBeAg) and for genotyping [15,16,17,18,19]. Most of these studies did not assess all the HBV markers on the same card and they generally did not use commercial assays. The aim was to investigate the performance of DBS in hepatitis B virus (HBV) diagnosis using commercial tests in comparison to standard methods
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