Comparison of Performance Characteristics of DxN VERIS System versus Qiagen PCR for HBV Genotype D and HCV Genotype 1b Quantification

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Polish Journal of Microbiology

Polish Society of Microbiologists

Subject: Microbiology

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ISSN: 1733-1331
eISSN: 2544-4646

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VOLUME 68 , ISSUE 1 (Mar 2019) > List of articles

Comparison of Performance Characteristics of DxN VERIS System versus Qiagen PCR for HBV Genotype D and HCV Genotype 1b Quantification

MURAT SAYAN / AYSE ARIKAN * / TAMER SANLIDAG

Citation Information : Polish Journal of Microbiology. VOLUME 68 , ISSUE 1 , ISSN (Online) 2544-4646, DOI: 10.21307/pjm-2019-008, Mar 2019

License : (CC-BY-NC-ND 4.0)

Received Date : 17-September-2018 / Accepted: 14-November-2018 / Published Online: 27-March-2019

ARTICLE

ABSTRACT

The Beckman Coulter DxN VERIS system is a fully automated, closed molecular diagnostic instrument for viral load quantification of hepatitis B virus and hepatitis C virus. In this study, the analytical performance of this new system was compared to routine diagnostic Qiagen PCR kit by using the same clinical samples. The DxN VERIS system demonstrated a high analytical performance. The DxN VERIS allows random access, which means that samples can be uploaded straight on to the system at any time; so, it provides an improvement of workflow, staff productivity and allows faster turn-around of viral load results.

Graphical ABSTRACT

Hepatitis B virus (HBV) and hepatitis C virus (HCV) are hepatotropic, non-cytopathic viruses able to establish a persistent infection that causes different degrees of hepatic inflammation (chronic hepatitis), leading to the development of liver cirrhosis and hepatocellular carcinoma (HCC) (Caccamo et al. 2014; Easterbrook et al. 2017). Today, the main goal of HBV therapy is to prevent the progression of the disease, improving survival and the quality of life; whereas, for the HCV therapy, the aim is to cure the infection to achieve a sustained virological response and consequently, to prevent HCC development (Akhan et al. 2015). Additionally, viral load (VL) monitoring is important in assessing the therapeutic response, monitoring treatment success and detecting drug-resistant viruses (Singh et al. 2017). There are several commercially available HBV-HCV monitoring assays which are mainly based on real-time polymerase chain reaction (RT-PCR); but, using the most effective, the cheapest and the fastest detecting system plays an important role in diagnosing viral infections. Although these techniques are preferred due to their excellent analytical sensitivity, specificity, accuracy, and broad dynamic range of linear quantification, they require many steps and consumables in the qualified laboratories (Portilho et al. 2015; Wu et al. 2017). VL assays also need to be batched as they arrive in the laboratory. Even a small change in the efficiency of the amplification can lead to striking differences in the amount final product. These reasons indicate the necessity of inventing fully standardized, reproducible and sensitive new assays for monitoring of HBV and HCV infections.

The aim of the study was to compare the analytical performance of random access testing (Beckman Coulter DxN VERIS) with routine diagnostic PCR kit (artus Qiagen).

A total of 67 chronic hepatitis B (CHB) patients with genotype D and 44 chronic hepatitis C (CHC) patients infected with genotype 1b from Kocaeli province in Marmara region of Turkey were analyzed in parallel with the use of DxN VERIS and Qiagen systems synchronously. Qiagen HBV DNA and HCV RNA levels in the samples were in the range from negative to > 108 IU/ml and > 107 IU/ml, respectively, and they are displayed in Table I. The PCR-negative samples were previously diagnosed as taken from the HBV and HCV carriers and their negativities were provided by antiviral treatment. The most predominant genotypes in Turkey are HBV genotype D and HCV genotype 1b; therefore, we have only involved these genotypes in our study (Karatas et al. 2018). The selected blood samples were immediately centrifuged, aliquoted, and then, stored at −80°C until use. The diagnosis of chronic hepatitis B and C was made on the basis of biochemical, serological, virological and histological data according to the EASL guidelines (EASL 2017a, 2017b). The samples were analyzed by using both fully automated DxN VERIS kits (Beckman Coulter DxN VERIS HBV/ HCV kits, Nyon, Switzerland) and Qiagen kits (artus HBV/HCV QS RGQ kit, Qiagen, Hilden, Germany) on the same day according to the manufacturer’s recommendations. DxN VERIS HBV/HCV assays have quantification ranges from 10–109 IU/ml and 12–108 IU/ml, respectively as it is stated in the package inserts. The relationship between VL quantifications measured by both DxN VERIS and Qiagen assays were analyzed by Passing-Bablok (PB) regression method. Bland Altman plot (BAP) design was used to calculate statistical limits, correlation and standard deviation (SD) of the differences between two types of measurement. To control the distribution of the differences and other properties between quantitative values, a graphical approach was used. Statistical analysis of the study was made with the Analyse-it Software program (Analyse-it Ltd. v 4.60, Microsoft Corp. Leeds, UK).

Table I

The Qiagen HBV DNA and HCV RNA levels in sampling.

10.21307_pjm-2019-008-tbl1.jpg

According to HBV BAP (DxN VERIS combined Qiagen HBV log IU/ml), the specificity resulted in 95% LoA (limits of agreement) [CL (confidence level) 95% = −1.59–0.61], the correlation was equal to 0.97 with SD 0.55 and a mean SD = −0.47. HBV PB indicated DxN VERIS combined = −1.179 + 1.153 Qiagen log IU/ml with the correlation equal to 0.97. HBV plots for PB and BAP are shown in Fig. 1.

Fig. 1.

HBV plots for Passing-Bablok (upper) and Bland Altman analysis (lower).

10.21307_pjm-2019-008-f001.jpg

For HCV analysis, BAP (DxN VERIS combined Qiagen HCV log IU/ml) illustrated the specificity of 95% LoA (CL 95% = −1.59−0.72), the correlation of 0.90 with SD 0.59 and a mean SD = −0.43. HCV PB analysis indicated DxN VERIS combined = −0.3394 + 0.8602 Qiagen log IU/ml with the correlation of 0.90. HCV plots for PB and BAP are shown in Fig. 2.

Fig. 2.

HCV plots for Passing-Bablok (upper) and Bland Altman analysis (lower).

10.21307_pjm-2019-008-f002.jpg

Viral nucleic acid detection is the gold standard for the detection of viral genomes in clinical samples. COBAS Ampliprep, artus Qiagen and Abbott real-time PCR assays are currently the most frequently used platforms worldwide in this field. DxN VERIS systems were mainly compared to COBAS and Abbott systems rather than to Qiagen kits (Saune et al. 2016; Patrick et al. 2017; Park et al. 2019). Studies comparing DxN VERIS HBV to Roche HBV and to Abbott real-time HBV illustrate the average bias that was determined as being equal to −0.26 (95% Cl: −0.37 to −0.15) and −0.36 (95% Cl: −0.43 to −0.29), respectively in BAP. This was equal to −0.32 log IU/ml in another study (Robert et al. 2014), and respectively to −0.47 log IU/ml in our study (Williams et al. 2014; Park et al. 2019). In the present study, we compared DxN VERIS with Qiagen HBV and HCV and their clinical performances displayed a strong correlation (95%) similar to one another study (Micheli et al. 2016). Currently, the main therapy is based on Peg-IFN and nucleos(t)ide analogues for CHB. However, the main clinical challenge is the development of antiviral resistance, since long-term therapeutic regimens are given to the majority of these patients (EASL 2017b). While monitoring VL, HBV breakthrough that is demonstrated by a sudden increase in DNA level (from > 1 log10 IU/ml to > 1 × 105 IU/ml) can be observed (Braun et al. 2017). Early detection of such viral breakthrough is important because the analysis of the HBV gene mutations responsible for drug resistance is a part of managing clinical treatment. The random access testing can provide effective surveillance for CHB treatment. Similarly, monitoring of HCV RNA during treatment is required to assess the success of the treatment and to detect any breakthrough related to viral resistance. As the main goal of therapy for HCV is to cure the infection in order to prevent the development of HCC and complications related with HCV related liver diseases, random access system enables faster turn-around of VL results to physicians and allows early detection of possible resistance. Therefore, DxN VERIS system may be a new solution as it enables same-day turn-around results to help the health system improving patient management. None of the other platforms are true single-sample to answer and random-access testing. This is the most important advantage of DxN VERIS system for us because medical reports can be transformed within a short time to different departments and physicians that could better manage the diseases with antiviral therapy in a short time.

Acknowledgments

The authors are thankful to Dr. Ferdiye Taner for English editing.

Ethical approved

The study has been approved by the Clinical Research Ethics Committee of Kocaeli University (KKAEK 2011/104).

Conflict of interest

The authors do not report any financial or personal connections with other persons or organizations, which might negatively affect the contents of this publication and/or claim authorship rights to this publication.

References


  1. Akhan S, Aynioglu A, Cagatay A, Gonen I, Gunal O, Kaynar T, Kuruuzum Z, Sayan M, Tunca B, Tulek N, et al. [Management of chronic hepatitis B virus infection: A consensus report of the study group for viral hepatitis of the Turkish society of clinical microbiology and infectious diseases]. (in Turkish). Klimik Dergisi/Klimik J. 2015;27(1):2–18. doi:10.5152/kd.2014.26
    [CROSSREF]
  2. Braun P, Drago M, Fanti D, Fleury H, Hofmann J, Izopet J, Kühn S, Lombardi A, Micheli V, Sauné K, et al. A European multicentre study on the comparison of HCV viral loads between VERIS HCV assay and COBAS® TaqMan® HCV Test and RealTime HCV Assay. J Clin Virol. 2017;90:18–25. doi:10.1016/j.jcv.2017.03.006 Medline
    [CROSSREF]
  3. Caccamo G, Saffioti F, Raimondo G. Hepatitis B virus and hepatitis C virus dual infection. World J Gastroenterol. 2014;20(40):14559–14567. doi:10.3748/wjg.v20.i40.14559 Medline
    [CROSSREF]
  4. Easterbrook PJ, Roberts T, Sands A, Peeling R. Diagnosis of viral hepatitis. Curr Opin HIV AIDS. 2017;12(3):302–314. doi:10.1097/COH.0000000000000370 Medline
    [CROSSREF]
  5. EASL [European Association for the Study of the Liver]. EASL recommendations on treatment of hepatitis C 2016. J Hepatol. 2017a; 66(1):153–194. doi:10.1016/j.jhep.2016.09.001
  6. EASL [European Association for the Study of the Liver]. EASL 2017 clinical practice guidelines on the management of hepatitis B virus infection. J Hepatol. 2017b;67(2):370–398. doi:10.1016/j.jhep.2017.03.021
  7. Karatas E, Erensoy S, Akarca US, Sertöz R. [Analysis of hepatitis B virus (HBV) preS1, preS2 and S gene regions from patient groups infected with HBV genotype D] (in Turkish). Mikrobiyol Bul. 2018; 52(1):23–34. Medline
    [CROSSREF]
  8. Micheli V, Lombardi A, Mancon A, Mileto D, Zanchetta N, Magni CF, Milazzo L, Angeli E, Gubertini G, Galli M, et al. DxN VERIS system for HCV viral load monitoring in the era of second wave direct-acting antiviral agents. J Hepatol. 2016;64(2):S627. doi:10.1016/S0168–8278(16)01165-X
    [CROSSREF]
  9. Park J, Cho H, Choi SJ, Lee GD, Sin SH, Ryu JH, Park HS, Lee H, Kim Y, Oh EJ. Performance evaluation of the Beckman Coulter DxN VERIS hepatitis B virus (HBV) assay in comparison with the Abbott real time HBV assay. Ann Lab Med. 2019;39(1):86–90. doi:10.3343/alm.2019.39.1.86 Medline
    [CROSSREF]
  10. Portilho MM, Baptista ML, da Silva M, de Sousa PSF, Lewis-Ximenez LL, Lampe E, Villar LM. Usefulness of in-house PCR methods for hepatitis B virus DNA detection. J Virol Methods. 2015;223:40–44. doi:10.1016/j.jviromet.2015.07.010 Medline
    [CROSSREF]
  11. Sauné K, Abravanel F, Haslé C, Boineau J, Mengelle C, Izopet J. Analytical performance of the VERIS MDx system HCV assay for detecting and quantifying HCV RNA. J Clin Virol. 2016;84:7–11. doi:10.1016/j.jcv.2016.09.003 Medline
    [CROSSREF]
  12. Singh MP, Galhotra S, Saigal K, Kumar A, Ratho RK. Quantitative nucleic acid amplification methods and their implications in clinical virology. Int J Appl Basic Med Res. 2017;7(1):3–9. doi:10.4103/2229–516X.198498 Medline
    [CROSSREF]
  13. Williams JA, Rodriguez J, Wang Z, Lykstad K, Donvito G, Fung J, Howell C, Colinayo VV, Gustafson E, Scholl T. Quantitative detection of hepatitis B Virus (HBV) on the VERIS MDx System. In: 24th European Congress of Clinical Microbiology and Infectious Diseases, 10–13 May 2014, Barcelona, Spain. [Internet] [cited 2018 September 14]; Poster Presentation:P1422. Available from: https://www.escmid.org/escmid_publications/escmid_elibrary/material/?mid=16949
  14. Wu B, Xiao F, Li P, Du Y, Lin J, Ming K, Chen B, Lei X, Xu B, Liu D. Ultrasensitive detection of serum hepatitis B virus by coupling ultra-filtration DNA extraction with real-time PCR. PLoS One. 2017; 12(2):e0170290. doi:10.1371/journal.pone.0170290 Medline
    [CROSSREF]
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FIGURES & TABLES

Fig. 1.

HBV plots for Passing-Bablok (upper) and Bland Altman analysis (lower).

Full Size   |   Slide (.pptx)

Fig. 2.

HCV plots for Passing-Bablok (upper) and Bland Altman analysis (lower).

Full Size   |   Slide (.pptx)

REFERENCES

  1. Akhan S, Aynioglu A, Cagatay A, Gonen I, Gunal O, Kaynar T, Kuruuzum Z, Sayan M, Tunca B, Tulek N, et al. [Management of chronic hepatitis B virus infection: A consensus report of the study group for viral hepatitis of the Turkish society of clinical microbiology and infectious diseases]. (in Turkish). Klimik Dergisi/Klimik J. 2015;27(1):2–18. doi:10.5152/kd.2014.26
    [CROSSREF]
  2. Braun P, Drago M, Fanti D, Fleury H, Hofmann J, Izopet J, Kühn S, Lombardi A, Micheli V, Sauné K, et al. A European multicentre study on the comparison of HCV viral loads between VERIS HCV assay and COBAS® TaqMan® HCV Test and RealTime HCV Assay. J Clin Virol. 2017;90:18–25. doi:10.1016/j.jcv.2017.03.006 Medline
    [CROSSREF]
  3. Caccamo G, Saffioti F, Raimondo G. Hepatitis B virus and hepatitis C virus dual infection. World J Gastroenterol. 2014;20(40):14559–14567. doi:10.3748/wjg.v20.i40.14559 Medline
    [CROSSREF]
  4. Easterbrook PJ, Roberts T, Sands A, Peeling R. Diagnosis of viral hepatitis. Curr Opin HIV AIDS. 2017;12(3):302–314. doi:10.1097/COH.0000000000000370 Medline
    [CROSSREF]
  5. EASL [European Association for the Study of the Liver]. EASL recommendations on treatment of hepatitis C 2016. J Hepatol. 2017a; 66(1):153–194. doi:10.1016/j.jhep.2016.09.001
  6. EASL [European Association for the Study of the Liver]. EASL 2017 clinical practice guidelines on the management of hepatitis B virus infection. J Hepatol. 2017b;67(2):370–398. doi:10.1016/j.jhep.2017.03.021
  7. Karatas E, Erensoy S, Akarca US, Sertöz R. [Analysis of hepatitis B virus (HBV) preS1, preS2 and S gene regions from patient groups infected with HBV genotype D] (in Turkish). Mikrobiyol Bul. 2018; 52(1):23–34. Medline
    [CROSSREF]
  8. Micheli V, Lombardi A, Mancon A, Mileto D, Zanchetta N, Magni CF, Milazzo L, Angeli E, Gubertini G, Galli M, et al. DxN VERIS system for HCV viral load monitoring in the era of second wave direct-acting antiviral agents. J Hepatol. 2016;64(2):S627. doi:10.1016/S0168–8278(16)01165-X
    [CROSSREF]
  9. Park J, Cho H, Choi SJ, Lee GD, Sin SH, Ryu JH, Park HS, Lee H, Kim Y, Oh EJ. Performance evaluation of the Beckman Coulter DxN VERIS hepatitis B virus (HBV) assay in comparison with the Abbott real time HBV assay. Ann Lab Med. 2019;39(1):86–90. doi:10.3343/alm.2019.39.1.86 Medline
    [CROSSREF]
  10. Portilho MM, Baptista ML, da Silva M, de Sousa PSF, Lewis-Ximenez LL, Lampe E, Villar LM. Usefulness of in-house PCR methods for hepatitis B virus DNA detection. J Virol Methods. 2015;223:40–44. doi:10.1016/j.jviromet.2015.07.010 Medline
    [CROSSREF]
  11. Sauné K, Abravanel F, Haslé C, Boineau J, Mengelle C, Izopet J. Analytical performance of the VERIS MDx system HCV assay for detecting and quantifying HCV RNA. J Clin Virol. 2016;84:7–11. doi:10.1016/j.jcv.2016.09.003 Medline
    [CROSSREF]
  12. Singh MP, Galhotra S, Saigal K, Kumar A, Ratho RK. Quantitative nucleic acid amplification methods and their implications in clinical virology. Int J Appl Basic Med Res. 2017;7(1):3–9. doi:10.4103/2229–516X.198498 Medline
    [CROSSREF]
  13. Williams JA, Rodriguez J, Wang Z, Lykstad K, Donvito G, Fung J, Howell C, Colinayo VV, Gustafson E, Scholl T. Quantitative detection of hepatitis B Virus (HBV) on the VERIS MDx System. In: 24th European Congress of Clinical Microbiology and Infectious Diseases, 10–13 May 2014, Barcelona, Spain. [Internet] [cited 2018 September 14]; Poster Presentation:P1422. Available from: https://www.escmid.org/escmid_publications/escmid_elibrary/material/?mid=16949
  14. Wu B, Xiao F, Li P, Du Y, Lin J, Ming K, Chen B, Lei X, Xu B, Liu D. Ultrasensitive detection of serum hepatitis B virus by coupling ultra-filtration DNA extraction with real-time PCR. PLoS One. 2017; 12(2):e0170290. doi:10.1371/journal.pone.0170290 Medline
    [CROSSREF]

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