Commercial Real-Time Reverse Transcriptase PCR Assays Can Underestimate or Fail to Quantify Hepatitis Delta Virus Viremia

Brichler, Ségolène; Gal, Frédéric Le; Butt, Afifaa; Chevret, Sylvie; Gordien, Emmanuel

doi:10.1016/j.cgh.2013.01.025

Cited by 44 publications

(36 citation statements)

References 25 publications

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“…Particularly, African HDV genotypes (i.e., HDV-1 and HDV-5 to -8) are poorly quantified by almost all available assays and are either undetected or dramatically underestimated (11). These different genotypes originally had a specific geographic distribution; however, due to ancient or recent migrations of populations, this genetic variability is now encountered in different countries worldwide (9)(10)(11)(17)(18)(19)(20)(21)(22)(23)(24)(25). In this respect, France, where genotypes HDV-5 to -8 representing 20% of spreading strains were characterized, is an outstanding example (26,27).…”

Section: Discussionmentioning

confidence: 99%

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Performance Characteristics of a New Consensus Commercial Kit for Hepatitis D Virus RNA Viral Load Quantification

et al. 2017

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Hepatitis D virus (HDV) is responsible for fulminant hepatitis and liver failure and accelerates evolution toward cirrhosis and hepatocellular carcinoma in hepatitis B virus (HBV)-infected patients. To date, treatment relies upon long-term administration of pegylated alpha-interferon with a sustained virological response in 30% of the patients. Very recently, new, promising anti-HDV therapies have been developed and are already being used in clinical trials. HDV RNA viral load (HDVL) monitoring must be an integral part of the management of the infected patients. However, HDV genus is characterized by a high genetic variability into eight genotypes (HDV-1 to -8), and most available in-house or commercial assays are useful for only a limited subset of genotypes. Results of a comparison of the performance of a new kit for HDVL quantification with the consensus in-house assay of the French National Reference Laboratory for HDV developed in 2005 are reported here. A total of 611 clinical samples of all HDV genotypes with various HDVL values, including several consecutive samples over several years from 36 patients, were studied. A specificity, sensitivity, and reproducibility evaluation was conducted using HDV-positive clinical samples, hepatitis A, B, C and E (HAV, HBV, HCV, and HEV, respectively) and HIV mono-infected samples, and the WHO HDV RNA international standard. Overall results were strictly comparable between the two assays (median difference, 0.07 log IU/ml), with high diagnosis precision and capacity. In summary, this new kit showed high performance in detection/quantification of HDVL, regardless of the genotype of the infecting strain used, and seems to be a suitable tool for patient management.

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Section: Discussionmentioning

confidence: 99%

“…However, very recently we showed that most of them dramatically underesti-mated or failed to detect/quantify positive HDV RNA samples, especially from patients infected with strains of African origin (HDV-1 and HDV-5 to -8) (9)(10)(11), highlighting the lack of efficient tools to routinely monitor HDVL for therapeutic management of infected patients.…”

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Performance Characteristics of a New Consensus Commercial Kit for Hepatitis D Virus RNA Viral Load Quantification

et al. 2017

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“…A few commercial methods are now available to quantify HDV RNA; but, as was recently reported by Brichler et al (7), they appear to underquantify non-genotype 1 sample. Several in-house methods based on quantitative reverse transcription-PCR (qRT-PCR) and targeting the HDAg or the ribozyme domain have been described and have proven effective for quantifying all HDV genotypes.…”

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confidence: 96%

Relevance of a Full-Length Genomic RNA Standard and a Thermal-Shock Step for Optimal Hepatitis Delta Virus Quantification

et al. 2014

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Hepatitis D virus (HDV) is a defective RNA virus that requires the surface antigens of hepatitis B virus (HBV) (HBsAg) for viral assembly and replication. Several commercial and in-house techniques have been described for HDV RNA quantification, but the methodologies differ widely, making a comparison of the results between studies difficult. In this study, a full-length genomic RNA standard was developed and used for HDV quantification by two different real-time PCR approaches (fluorescence resonance energy transfer [FRET] and TaqMan probes). Three experiments were performed. First, the stability of the standard was determined by analyzing the effect of thawing and freezing. Second, because of the strong internal base pairing of the HDV genome, which leads to a rod-like structure, the effect of intense thermal shock (95°C for 10 min and immediate cooling to ؊80°C) was tested to confirm the importance of this treatment in the reverse transcription step. Lastly, to investigate the differences between the DNA and RNA standards, the two types were quantified in parallel with the following results: the full-length genomic RNA standard was stable and reliably mimicked the behavior of HDV-RNA-positive samples, thermal shock enhanced the sensitivity of HDV RNA quantification, and the DNA standard underquantified the HDV RNA standard. These findings indicate the importance of using complete full-length genomic RNA and a strong thermal-shock step for optimal HDV RNA quantification. Hepatitis D virus (HDV) is a small defective RNA virus that requires hepatitis B virus surface antigens (HBsAg) for viral assembly and replication. The HDV genome is a circular, negative, and single-stranded RNA composed of 1,672 to 1,697 nucleotides. HDV virions present a nucleocapsid-like structure formed by genomic HDV-RNA, which is associated with the hepatitis delta antigen (HDAg) protein and surrounded by the three envelope proteins of the hepatitis B virus (1). The replication of HDV occurs intrahepatically through a double-rolling circle mechanism that leads to an antigenomic RNA intermediate that is the exact complement sequence of the genome. The genomic and antigenomic molecules both present 70% internal base pairing, enabling self-assembly of the molecule and leading to a rod-like structure. An additional RNA form, the 800-nucleotide mRNA responsible for expression of the hepatitis delta antigen (HDAg), is also known to be produced during replication (2).HDV infection causes acute or chronic liver disease in patients with hepatitis B virus (HBV) infection. HDV infection is widespread, and the prevalence differs between areas, but it is estimated that among the 350 million individuals with chronic HBV infection, approximately 15 million are also exposed to HDV (2). Eight HDV genotypes with specific geographic patterns have been reported (3). Nonetheless, population migration has led to changes in the distributions of HDV infection and genotypes (4-6).The diagnosis of HDV infection relies on the detection of specific antibodies (anti...

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“…An immune-prophylaxis, as already cited, is justified to decrease the risk of HBV relapse after transplantation. The 5-year survival rate is around 90% [18]. When hepatocellular carcinoma is established, the best therapeutic strategy may be the liver transplantation that cures the tumor and the cirrhosis, with a low risk of tumor recurrence at 2 years if the tumor size is small (<3 cm).…”

Section: Chronic B-delta Hepatitismentioning

confidence: 99%

Hepatitis Delta Virus: Epidemiology, Natural Course and Treatment

Sultanik¹,

Pol²

2016

J Infect Dis Ther

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The hepatitis delta virus (HDV) is a defective hepatotropic virus that affects only patients with hepatitis B virus (HBV) infection. These 2 viruses share the same routes of transmission (parenteral, sexual, and mother to child). The hepatitis delta virus infection may result in acute hepatitis, including fulminant presentation or spontaneously resolving infection, and chronic infection. The prognosis depends on the chronology of the 2 infections, co-infection (higher risk of fulminant hepatitis) or super-infection (frequent evolution to chronicity). The harmfull impact of HIVassociated infection is today debated even if HDV antibodies are present in 12% and 4% of HIV/HBV co-infected and HBV-mono-infected patients respectively. HDV may be treated by interferon, the unique treatment, but relapse is observed in 50% of cases after discontinuation of therapy: only 25% of treated patients achieved a sustained virologic response. Entry HBV/HDV inhibitors (Myrcludex) and prenylation inhibitors are awaited encouraging progress in treating HDV infection. The treatment is recommended in patients with significant fibrosis; viral suppression may result in fibrosis stabilization or reversal which may be also observed in the absence of complete viral eradication. The prevention of hepatitis delta virus infection is based on hepatitis B virus vaccination

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Commercial Real-Time Reverse Transcriptase PCR Assays Can Underestimate or Fail to Quantify Hepatitis Delta Virus Viremia

Cited by 44 publications

References 25 publications

Performance Characteristics of a New Consensus Commercial Kit for Hepatitis D Virus RNA Viral Load Quantification

Performance Characteristics of a New Consensus Commercial Kit for Hepatitis D Virus RNA Viral Load Quantification

Relevance of a Full-Length Genomic RNA Standard and a Thermal-Shock Step for Optimal Hepatitis Delta Virus Quantification

Hepatitis Delta Virus: Epidemiology, Natural Course and Treatment

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