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HBV-RNA Co-amplification May Influence HBV DNA Viral Load Determination.
Maasoumy, Benjamin Geretti, Anna Maria Frontzek, André Austin, Harrison Aretzweiler, Gudrun Garcia-Álvarez, Monica Leuchter, Susanne Simon, Christian O Marins, Ed G Canchola, Jesse A
Maasoumy, Benjamin
Geretti, Anna Maria
Frontzek, André
Austin, Harrison
Aretzweiler, Gudrun
Garcia-Álvarez, Monica
Leuchter, Susanne
Simon, Christian O
Marins, Ed G
Canchola, Jesse A
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2020-05-26
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Abstract
Despite effective hepatitis B virus (HBV)-DNA suppression, HBV RNA can circulate in patients receiving nucleoside/nucleotide analogues (NAs). Current assays quantify HBV DNA by either real-time polymerase chain reaction (PCR), which uses DNA polymerase, or transcription-mediated amplification, which uses reverse-transcriptase (RT) and RNA polymerase. We assessed the effect of RT capability on HBV-DNA quantification in samples from three cohorts, including patients with quantified HBV RNA. We compared the HBV-DNA levels by real-time PCR (cobas HBV, Roche 6800/8800; Xpert HBV, Cepheid), transcription-mediated amplification (Aptima HBV, Hologic), and real-time PCR with added RT capability (cobas HBV+RT). In the first cohort (n = 45) followed over 192 weeks of NA therapy, on-treatment HBV-DNA levels were higher with cobas HBV+RT than cobas HBV (mean difference: 0.14 log10 IU/mL). In a second cohort (n = 50) followed over 96 weeks of NA therapy, HBV-DNA viral load was significantly higher with the cobas HBV+RT and Aptima HBV compared with the cobas HBV test at all time points after initiation of NA therapy (mean difference: 0.65-1.16 log10 IU/mL). A clinically significant difference was not detected between the assays at baseline. In a third cohort (n = 53), after a median of 2.2 years of NA therapy, we detected HBV RNA (median 5.6 log10 copies/mL) in 23 patients (43.4%). Median HBV-DNA levels by Aptima HBV were 2.4 versus less than 1 log10 IU/mL in samples with HBV RNA and without HBV RNA, respectively (P = 0.0006). In treated patients with HBV RNA, Aptima HBV measured higher HBV-DNA levels than Xpert HBV and cobas HBV. Conclusion: Tests including an RT step may overestimate HBV DNA, particularly in samples with low viral loads as a result of NA therapy. This overestimation is likely due to amplification of HBV RNA and may have an impact on clinical decisions.
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Hepatol Commun. 2020 May 26;4(7):983-997. doi: 10.1002/hep4.1520. PMID: 32626831.
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en
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2471-254X
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Attribution 4.0 International