Method for Quantitative HDV RNA Detection: I, Manual Workflow (Serum and Liver Tissue) and II, Fully Automated High Throughput Workflow for Diagnostic Use
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Method for Quantitative HDV RNA Detection: I, Manual Workflow (Serum and Liver Tissue) and II, Fully Automated High Throughput Workflow for Diagnostic Use. / Pflüger, Lisa Sophie; Volz, Tassilo; Giersch, Katja; Allweiss, Lena; Dandri-Petersen, Maura; Lütgehetmann, Marc.
in: Methods Mol Biol, Jahrgang 2837, 2024, S. 171-184.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Method for Quantitative HDV RNA Detection: I, Manual Workflow (Serum and Liver Tissue) and II, Fully Automated High Throughput Workflow for Diagnostic Use
AU - Pflüger, Lisa Sophie
AU - Volz, Tassilo
AU - Giersch, Katja
AU - Allweiss, Lena
AU - Dandri-Petersen, Maura
AU - Lütgehetmann, Marc
N1 - © 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2024
Y1 - 2024
N2 - The hepatitis delta virus (HDV) is a small RNA virus (1700 base pairs), which uses the surface proteins of the hepatitis B virus (HBV) as an envelope. Accurate and reliable quantitative detection of HDV RNA is central for scientific and translational clinical research or diagnostic purposes. However, HDV poses challenges for nucleic acid amplification techniques: (1) the circular genome displays high intramolecular base pairing; (2) high content of cytosine and guanine; and (3) enormous genomic diversity among the eight known HDV genotypes (GTs). Here, we provide step-by-step instructions for (A) a manual workflow to perform a quantitative HDV reverse transcription (RT)-PCR from serum and liver tissue and (B) a quantitative HDV RT-PCR assay with whole process control to be used for serum or plasma samples run on a fully automated system. Both assays target the conserved ribozyme region and demonstrate inclusivity for all eight HDV GTs. The choice of assay depends on the experimental needs and equipment availability. While the former is ideal for scientific research laboratories, the latter provides a useful tool in the field of translational research or diagnostics.
AB - The hepatitis delta virus (HDV) is a small RNA virus (1700 base pairs), which uses the surface proteins of the hepatitis B virus (HBV) as an envelope. Accurate and reliable quantitative detection of HDV RNA is central for scientific and translational clinical research or diagnostic purposes. However, HDV poses challenges for nucleic acid amplification techniques: (1) the circular genome displays high intramolecular base pairing; (2) high content of cytosine and guanine; and (3) enormous genomic diversity among the eight known HDV genotypes (GTs). Here, we provide step-by-step instructions for (A) a manual workflow to perform a quantitative HDV reverse transcription (RT)-PCR from serum and liver tissue and (B) a quantitative HDV RT-PCR assay with whole process control to be used for serum or plasma samples run on a fully automated system. Both assays target the conserved ribozyme region and demonstrate inclusivity for all eight HDV GTs. The choice of assay depends on the experimental needs and equipment availability. While the former is ideal for scientific research laboratories, the latter provides a useful tool in the field of translational research or diagnostics.
KW - Hepatitis Delta Virus/genetics
KW - Humans
KW - RNA, Viral/genetics
KW - Workflow
KW - Hepatitis D/diagnosis
KW - Liver/virology
KW - Reverse Transcriptase Polymerase Chain Reaction/methods
KW - Real-Time Polymerase Chain Reaction/methods
KW - Genotype
U2 - 10.1007/978-1-0716-4027-2_15
DO - 10.1007/978-1-0716-4027-2_15
M3 - SCORING: Journal article
C2 - 39044084
VL - 2837
SP - 171
EP - 184
JO - Methods Mol Biol
JF - Methods Mol Biol
SN - 1064-3745
ER -
