Home Practical approach to method verification in plasma and validation in cerebrospinal fluid under accreditation using a flexible scope in molecular virology: setting up the HIV, HBV and HCV Aptima™ Quant Dx assays
Article
Licensed
Unlicensed Requires Authentication

Practical approach to method verification in plasma and validation in cerebrospinal fluid under accreditation using a flexible scope in molecular virology: setting up the HIV, HBV and HCV Aptima™ Quant Dx assays

  • Caroline Lefeuvre , Adeline Pivert , Cong T. Tran , Françoise Lunel-Fabiani , Alexandra Ducancelle and Hélène Le Guillou-Guillemette EMAIL logo
Published/Copyright: September 20, 2019
Become an author with De Gruyter Brill
Submit Manuscript Author Information
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 58 Issue 2

Abstract

Background

Our laboratory obtained the ISO 15189 accreditation for the plasmatic HIV-1, HBV and HCV viral load (VL) using the m2000 RealTime™ system, which was recently changed for the platform Panther®. Here, we discuss a strategy for performing method validation/verification very quickly.

Methods

We performed the mandatory (repeatability, internal quality assessment [IQA], measurement uncertainty [MU]) and optional technical verifications for CE/IVD assays using the flexible scope range A. We also performed the mandatory assays for the validation of HIV-1 VL in the cerebrospinal fluid (CSF) using the flexible scope range B. The change was checked by following up on the turnaround time (TAT).

Results

The coefficient of variation (CV%) for repeatability and IQA complied with the limit of 0.25 log. The MU results ranged from 0.04 to 0.25 log copies or IU/mL. The comparisons of methods showed excellent correlations (R2 = 0.96 for the three parameters) but a delayed centrifugation on HCV VL showed variations of up to 2 log IU/mL. An excellent linearity for HIV-1 in the CSF was obtained from 1.5 to 5 log copies/mL with R2 = 0.99. The TAT increased (84%–98%) in routine usage.

Conclusions

The three Aptima assays are well suited for routine laboratory use and can be integrated within less than 2 weeks in accordance with flexible scope range A. Our data allows us to confidently perform HIV-1 VL in CSF following flexible scope range B. Finally, we provide an organizational guide for flexible scope management in molecular virology within a short time frame.

Keywords: HBV; HCV; HIV-1; flexible scope management; ISO 15189; method validation; method verification; viral load
Corresponding author: Hélène Le Guillou-Guillemette, PharmD, PhD, Virology Department, Angers University Hospital, HIFIH Laboratory EA 3859, LUNAM, 4, rue Larrey 49000 Angers, France, Phone: 02 41 35 47 09

Acknowledgments

The authors thank Mrs Rachael Barton for proofreading the English manuscript.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

References

1. International Organization for Standardization (ISO). Medical laboratories – requirements for quality and competence. http://www.iso.org/cms/render/live/en/sites/isoorg/contents/data/standard/05/61/56115.html. Accessed: July 2019.Search in Google Scholar

2. Padoan A, Sciacovelli L, Aita A, Antonelli G, Plebani M. Measurement uncertainty in laboratory reports: a tool for improving the interpretation of test results. Clin Biochem 2018;57:41–7.10.1016/j.clinbiochem.2018.03.009Search in Google Scholar PubMed

3. Plebani M, Sciacovelli L, Bernardi D, Aita A, Antonelli G, Padoan A. What information on measurement uncertainty should be communicated to clinicians, and how? Clin Biochem 2018;57:18–22.10.1016/j.clinbiochem.2018.01.017Search in Google Scholar PubMed

4. Padoan A, Antonelli G, Aita A, Sciacovelli L, Plebani M. An approach for estimating measurement uncertainty in medical laboratories using data from long-term quality control and external quality assessment schemes. Clin Chem Lab Med 2017;55:1696–701.10.1515/cclm-2016-0896Search in Google Scholar PubMed

5. Antonelli G, Padoan A, Aita A, Sciacovelli L, Plebani M. Verification of examination procedures in clinical laboratory for imprecision, trueness and diagnostic accuracy according to ISO 15189:2012: a pragmatic approach. Clin Chem Lab Med 2017;55:1501–8.10.1515/cclm-2016-0894Search in Google Scholar

6. Plebani M, Sciacovelli L, Aita A. Quality indicators for the total testing process. Clin Lab Med 2017;37:187–205.10.1016/j.cll.2016.09.015Search in Google Scholar PubMed

7. Comité Français d’Accréditation. SH GTA 01: Guide technique d’accréditation en biologie médicale. Révision 02. https://tools.cofrac.fr/documentation/SH-GTA-01. Accessed: July 2019.Search in Google Scholar

8. Comité Français d’Accréditation. SH GTA 04 : Guide technique d’accréditation de verification (portée A)/validation (portée B) des méthodes en biologie médicale. https://tools.cofrac.fr/documentation/SH-GTA-04. Accessed: Jul 2019.Search in Google Scholar

9. Comité Français d’Accréditation. SH GTA 14 : Guide technique d’accréditation pour l’évaluation des incertitudes de mesure en biologie médicale. https://tools.cofrac.fr/documentation/SH-GTA-14. Accessed: July 2019.Search in Google Scholar

10. Thelen MH, Vanstapel FJ, Kroupis C, Vukasovic I, Boursier G, Barrett E, et al. Flexible scope for ISO 15189 accreditation: a guidance prepared by the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) Working Group Accreditation and ISO/CEN Standards (WG-A/ISO). Clin Chem Lab Med 2015;53:1173–80.10.1515/cclm-2015-0257Search in Google Scholar PubMed

11. Schønning K, Pedersen MS, Johansen K, Landt B, Nielsen LG, Weis N, et al. Analytical and clinical performance of the Hologic Aptima HCV Quant Dx assay for the quantification of HCV RNA in plasma samples. J Virol Methods 2017;248:159–65.10.1016/j.jviromet.2017.07.006Search in Google Scholar PubMed

12. Hughes MD, Johnson VA, Hirsch MS, Bremer JW, Elbeik T, Erice A, et al. Monitoring plasma HIV-1 RNA levels in addition to CD4+ lymphocyte count improves assessment of antiretroviral therapeutic response. ACTG 241 Protocol Virology Substudy Team. Ann Intern Med 1997;126:929–38.10.7326/0003-4819-126-12-199706150-00001Search in Google Scholar PubMed

13. Saag MS, Holodniy M, Kuritzkes DR, O’Brien WA, Coombs R, Poscher ME, et al. HIV viral load markers in clinical practice. Nat Med 1996;2:625–9.10.1038/nm0696-625Search in Google Scholar PubMed

14. Antonelli G, Sciacovelli L, Aita A, Padoan A, Plebani M. Validation model of a laboratory-developed method for the ISO15189 accreditation: the example of salivary cortisol determination. Clin Chim Acta 2018;485:224–8.10.1016/j.cca.2018.07.005Search in Google Scholar PubMed

15. Pum J. A practical guide to validation and verification of analytical methods in the clinical laboratory. Adv Clin Chem 2019;90:215–81.10.1016/bs.acc.2019.01.006Search in Google Scholar PubMed

16. Thelen M, Vanstapel F, Brguljan PM, Gouget B, Boursier G, Barrett E, et al. Documenting metrological traceability as intended by ISO 15189:2012: a consensus statement about the practice of the implementation and auditing of this norm element. Clin Chem Lab Med 2019;57:459–64.10.1515/cclm-2018-1212Search in Google Scholar PubMed

17. Lamy B, Ferroni A, Henning C, Cattoen C, Laudat P. How to: accreditation of blood cultures’ proceedings. A clinical microbiology approach for adding value to patient care. Clin Microbiol Infect 2018;24:956–63.10.1016/j.cmi.2018.01.011Search in Google Scholar PubMed

18. Fihman V, Bleunven SC, Le Glaunec JM, Maillebuau F, De Rochebouet I, Nebbad-Lechani B, et al. Are bacterial culture quantifications reliable? Comparative performance of the WASP automated inoculation instrument in the era of ISO 15189 accreditation. J Med Microbiol 2018;67:1581–8.10.1099/jmm.0.000847Search in Google Scholar PubMed

19. Lafontaine S, Prin-Mathieu C, Velay A, Agulles O, Schvoerer E, Jeulin H. Evaluation of virological microparticle enzyme immunoassay according to the ISO 15189: real-life experience in a University Hospital. Clin Lab 2016;62:275–84.10.7754/Clin.Lab.2015.150214Search in Google Scholar

20. Bogaerts P, Rezende de Castro R, de Mendonça R, Huang T-D, Denis O, Glupczynski Y. Validation of carbapenemase and extended-spectrum β-lactamase multiplex endpoint PCR assays according to ISO 15189. J Antimicrob Chemother 2013;68:1576–82.10.1093/jac/dkt065Search in Google Scholar PubMed

21. Rabenau HF, Kessler HH, Kortenbusch M, Steinhorst A, Raggam RB, Berger A. Verification and validation of diagnostic laboratory tests in clinical virology. J Clin Virol 2007;40:93–8.10.1016/j.jcv.2007.07.009Search in Google Scholar PubMed

22. Boursier G, Vukasovic I, Brguljan PM, Lohmander M, Ghita I, Bernabeu Andreu FA, et al. Accreditation process in European countries – an EFLM survey. Clin Chem Lab Med 2016;54:545–51.10.1515/cclm-2015-0780Search in Google Scholar PubMed

23. May S, Adamska E, Tang JW. Evaluating the aptima HIV-1 quant Dx, HCV quant Dx and HBV quant assays against the Abbott HIV-1, HCV and HBV RealTime assays. J Clin Virol 2018;106:7–10.10.1016/j.jcv.2018.06.015Search in Google Scholar PubMed

24. Sauné K, Raymond S, Boineau J, Pasquier C, Izopet J. Detection and quantification of HIV-1 RNA with a fully automated transcription-mediated-amplification assay. J Clin Virol 2016;84:70–3.10.1016/j.jcv.2016.09.002Search in Google Scholar PubMed

25. Yek C, Massanella M, Peling T, Lednovich K, Nair SV, Worlock A, et al. Evaluation of the Aptima HIV-1 Quant Dx assay for HIV-1 RNA quantitation in different biological specimen types. J Clin Microbiol 2017;55:2544–53.10.1128/JCM.00425-17Search in Google Scholar PubMed PubMed Central

26. Manak MM, Eller LA, Malia J, Jagodzinski LL, Trichavaroj R, Oundo J, et al. Identification of Acute HIV-1 infection by hologic Aptima HIV-1 RNA qualitative assay. J Clin Microbiol 2017;55:2064–73.10.1128/JCM.00431-17Search in Google Scholar PubMed PubMed Central

27. Hatzakis A, Papachristou H, Nair SJ, Fortunko J, Foote T, Kim H, et al. Analytical characteristics and comparative evaluation of Aptima HIV-1 Quant Dx assay with Ampliprep/COBAS TaqMan HIV-1 test v2.0. Virol J 2016;13:176.10.1186/s12985-016-0627-ySearch in Google Scholar PubMed PubMed Central

28. Nair SV, Kim HC, Fortunko J, Foote T, Peling T, Tran C, et al. Aptima HIV-1 Quant Dx–a fully automated assay for both diagnosis and quantification of HIV-1. J Clin Virol 2016;77:46–54.10.1016/j.jcv.2016.02.002Search in Google Scholar PubMed

29. Schalasta G, Speicher A, Börner A, Enders M. Performance of the new Aptima HCV Quant Dx assay in comparison to the Cobas TaqMan HCV2 test for use with the high pure system in detection and quantification of hepatitis C Virus RNA in plasma or serum. J Clin Microbiol 2016;54:1101–7.10.1128/JCM.03236-15Search in Google Scholar PubMed PubMed Central

30. Chevaliez S, Dubernet F, Dauvillier C, Hézode C, Pawlotsky J-M. The new Aptima HCV Quant Dx real-time TMA assay accurately quantifies hepatitis C Virus Genotype 1–6 RNA. J Clin Virol 2017;91:5–11.10.1016/j.jcv.2017.03.020Search in Google Scholar PubMed

31. Chevaliez S, Dauvillier C, Dubernet F, Poveda J-D, Laperche S, Hézode C, et al. The new Aptima HBV Quant real-time TMA assay accurately quantifies hepatitis B Virus DNA from Genotypes A to F. J Clin Microbiol 2017;55:1211–9.10.1128/JCM.02219-16Search in Google Scholar PubMed PubMed Central

32. Canestri A, Lescure F-X, Jaureguiberry S, Moulignier A, Amiel C, Marcelin AG, et al. Discordance between cerebral spinal fluid and plasma HIV replication in patients with neurological symptoms who are receiving suppressive antiretroviral therapy. Clin Infect Dis 2010;50:773–8.10.1086/650538Search in Google Scholar PubMed

Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2019-0502).

Received: 2019-05-17
Accepted: 2019-08-22
Published Online: 2019-09-20
Published in Print: 2020-01-28

©2019 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Towards a personalized assessment of vitamin D status
  4. Reviews
  5. Circulating tumor DNA and their added value in molecular oncology
  6. Telomere length determinants in childhood
  7. Opinion Papers
  8. Serum or plasma? An old question looking for new answers
  9. Evaluating sample stability in the clinical laboratory with the help of linear and non-linear regression analysis
  10. General Clinical Chemistry and Laboratory Medicine
  11. Simultaneous measurement of 25(OH)-vitamin D and 24,25(OH)2-vitamin D to define cut-offs for CYP24A1 mutation and vitamin D deficiency in a population of 1200 young subjects
  12. How well do Croatian laboratories adhere to national recommendations for laboratory diagnostics of chronic kidney disease (CKD)?
  13. Underfilling of vacuum blood collection tubes leads to increased lactate dehydrogenase activity in serum and heparin plasma samples
  14. Calcium state estimation by total calcium: the evidence to end the never-ending story
  15. The use of faecal immunochemical testing in the decision-making process for the endoscopic investigation of iron deficiency anaemia
  16. Measurement uncertainty of β-lactam antibiotics results: estimation and clinical impact on therapeutic drug monitoring
  17. Practical approach to method verification in plasma and validation in cerebrospinal fluid under accreditation using a flexible scope in molecular virology: setting up the HIV, HBV and HCV Aptima™ Quant Dx assays
  18. Plasma neurofilament light chain is associated with mortality after spontaneous intracerebral hemorrhage
  19. Comparison of the diagnostic performance of two automated urine sediment analyzers with manual phase-contrast microscopy
  20. Development and validation of LC-MS/MS methods to measure tobramycin and lincomycin in plasma, microdialysis fluid and urine: application to a pilot pharmacokinetic research study
  21. Reference Values and Biological Variations
  22. Cord blood S100B: reference ranges and interest for early identification of newborns with brain injury
  23. Hematology and Coagulation
  24. Validation and standardization of the ETP-based activated protein C resistance test for the clinical investigation of steroid contraceptives in women: an unmet clinical and regulatory need
  25. Cancer Diagnostics
  26. Next-generation sequencing for tumor mutation quantification using liquid biopsies
  27. Cardiovascular Diseases
  28. Evolution of the slopes of ST2 and galectin-3 during marathon and ultratrail running compared to a control group
  29. Letters to the Editor
  30. Biological variation of two serum markers for preeclampsia prediction
  31. Daily monitoring of a control material with a concentration near the limit of detection improves the measurement accuracy of highly sensitive troponin assays
  32. Are patients adequately informed about procedures for 24-h urine collection?
  33. Interferences in free thyroxine concentration using the Roche analytical platform: improvement of the third generation?
  34. Procedures for the diagnosis of macro-follicle stimulating hormone (FSH) in a patient with high serum FSH concentrations
  35. False-positive result of immunochromatographic (IC) strip test for the diagnosis of α-thalassemia in samples with autoantibodies
  36. Sample dilution for soluble interleukin-2 receptor α measurement: comparison of two different matrices
  37. The growing problem of predatory publishing: a case report
  38. Cerebrospinal fluid lactate levels according to the site of puncture
https://doi.org/10.1515/cclm-2019-0502
Keywords for this article
HBV; HCV; HIV-1; flexible scope management; ISO 15189; method validation; method verification; viral load

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Towards a personalized assessment of vitamin D status
  4. Reviews
  5. Circulating tumor DNA and their added value in molecular oncology
  6. Telomere length determinants in childhood
  7. Opinion Papers
  8. Serum or plasma? An old question looking for new answers
  9. Evaluating sample stability in the clinical laboratory with the help of linear and non-linear regression analysis
  10. General Clinical Chemistry and Laboratory Medicine
  11. Simultaneous measurement of 25(OH)-vitamin D and 24,25(OH)2-vitamin D to define cut-offs for CYP24A1 mutation and vitamin D deficiency in a population of 1200 young subjects
  12. How well do Croatian laboratories adhere to national recommendations for laboratory diagnostics of chronic kidney disease (CKD)?
  13. Underfilling of vacuum blood collection tubes leads to increased lactate dehydrogenase activity in serum and heparin plasma samples
  14. Calcium state estimation by total calcium: the evidence to end the never-ending story
  15. The use of faecal immunochemical testing in the decision-making process for the endoscopic investigation of iron deficiency anaemia
  16. Measurement uncertainty of β-lactam antibiotics results: estimation and clinical impact on therapeutic drug monitoring
  17. Practical approach to method verification in plasma and validation in cerebrospinal fluid under accreditation using a flexible scope in molecular virology: setting up the HIV, HBV and HCV Aptima™ Quant Dx assays
  18. Plasma neurofilament light chain is associated with mortality after spontaneous intracerebral hemorrhage
  19. Comparison of the diagnostic performance of two automated urine sediment analyzers with manual phase-contrast microscopy
  20. Development and validation of LC-MS/MS methods to measure tobramycin and lincomycin in plasma, microdialysis fluid and urine: application to a pilot pharmacokinetic research study
  21. Reference Values and Biological Variations
  22. Cord blood S100B: reference ranges and interest for early identification of newborns with brain injury
  23. Hematology and Coagulation
  24. Validation and standardization of the ETP-based activated protein C resistance test for the clinical investigation of steroid contraceptives in women: an unmet clinical and regulatory need
  25. Cancer Diagnostics
  26. Next-generation sequencing for tumor mutation quantification using liquid biopsies
  27. Cardiovascular Diseases
  28. Evolution of the slopes of ST2 and galectin-3 during marathon and ultratrail running compared to a control group
  29. Letters to the Editor
  30. Biological variation of two serum markers for preeclampsia prediction
  31. Daily monitoring of a control material with a concentration near the limit of detection improves the measurement accuracy of highly sensitive troponin assays
  32. Are patients adequately informed about procedures for 24-h urine collection?
  33. Interferences in free thyroxine concentration using the Roche analytical platform: improvement of the third generation?
  34. Procedures for the diagnosis of macro-follicle stimulating hormone (FSH) in a patient with high serum FSH concentrations
  35. False-positive result of immunochromatographic (IC) strip test for the diagnosis of α-thalassemia in samples with autoantibodies
  36. Sample dilution for soluble interleukin-2 receptor α measurement: comparison of two different matrices
  37. The growing problem of predatory publishing: a case report
  38. Cerebrospinal fluid lactate levels according to the site of puncture
Sign up now to receive a 20% welcome discount
Institutional Access
How does access work?
Have an idea on how to improve our website?
Please write us.
© 2025 De Gruyter Brill
Downloaded on 1.11.2025 from https://www.degruyterbrill.com/document/doi/10.1515/cclm-2019-0502/html
Scroll to top button