Z Gastroenterol 2019; 57(01): e83
DOI: 10.1055/s-0038-1677269
5. Viral Hepatitis, Immunology
Georg Thieme Verlag KG Stuttgart · New York

A rapid point-of-care device for the diagnosis of Hepatitis Delta Virus infection

FA Lempp
1   Department of Infectious Diseases, Molecular Virology, University Hospital of Heidelberg, Heidelberg, Germany
2   German Center for Infection Research (DZIF), partner site Heidelberg, Heidelberg, Germany
,
V Sakin
1   Department of Infectious Diseases, Molecular Virology, University Hospital of Heidelberg, Heidelberg, Germany
,
S Nkongolo
1   Department of Infectious Diseases, Molecular Virology, University Hospital of Heidelberg, Heidelberg, Germany
2   German Center for Infection Research (DZIF), partner site Heidelberg, Heidelberg, Germany
,
P Schnitzler
3   Department of Infectious Diseases, Virology, University Hospital of Heidelberg, Germany
,
H Wedemeyer
4   Clinic for Gastroenterology and Hepatology, University Hospital of Essen, Germany & German Centre for Infection Research (DZIF),
,
FL Gal
5   Laboratoire de Microbiologie Clinique, Hôpital Avicenne, APHP, Bobigny, France
,
E Gordien
5   Laboratoire de Microbiologie Clinique, Hôpital Avicenne, APHP, Bobigny, France
,
C Yurdaydin
6   Department of Gastroenterology, Ankara University, Ankara, Turkey
,
S Urban
1   Department of Infectious Diseases, Molecular Virology, University Hospital of Heidelberg, Heidelberg, Germany
2   German Center for Infection Research (DZIF), partner site Heidelberg, Heidelberg, Germany
› Author Affiliations
Further Information

Publication History

Publication Date:
04 January 2019 (online)

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Background:

Chronic Hepatitis Delta Virus (HDV) infection represents the most severe form of viral hepatitis. The detection of anti-HDV antibodies against Hepatitis Delta Antigen (HDAg) represents the recommended initial step in the diagnosis of HDV infection. Current assays for anti-HDV detection rely on manual ELISAs, which are time consuming and require laboratory equipment and trained staff. Our aim was to develop a rapid pan-genotypic point of care (POC) test that allows fast and reliable detection of anti-HDV antibodies for all eight HDV genotypes and that can be used in hospitals and doctors' practices but also in remote areas.

Methods:

Using multiple sequence alignments, we designed and optimized a pan-genotypic consensus sequence of HDAg. The corresponding recombinant protein was expressed, purified and implemented as a detection antigen to assemble a POC device, based on the principle of lateral flow assay. The POC device was validated with a panel of 332 HDV-positive and 142 HDV-negative patient samples. Additionally, 18 serum samples from patients with non-genotype 1 HDV infection were used to evaluate pan-genotypic test performance.

Results:

Application of serum containing anti-HDV antibodies to the POC test strips led to a visible positive test result already after 10 – 20 minutes of incubation. Positive signals could be detected even if the patient sera were diluted 100-fold, while no false positive signals were detected when HDV-negative control sera were applied. POC test validation on 476 samples led to a sensitivity of 94,6% and specificity of 100%. All 18 serum samples from non-genotype 1 HDV-infected patients were positive in the POC test indicating pan-genotypic sensitivity. Multiplexing the test strip with HBsAg-specific antibodies allowed simultaneous detection of HBsAg and HDV infection on the same strip.

Conclusion:

In light of the several novel HDV-specific drugs that are currently entering registration trials and might be available in the near future, fast and reliable diagnostics for HDV infection are of utmost importance. Here, we have developed and successfully validated a rapid test device for POC diagnosis in hospitals and remote areas or for epidemiological field studies.