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Meticulous plasma isolation is essential to avoid false low-level viraemia in Roche Cobas HIV-1 viral load assays

(2018) ANTIVIRAL THERAPY. 23(3). p.277-281
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Abstract
Background: Pre-analytical sample processing is often overlooked as a potential cause of inaccurate assay results. Here we demonstrate how plasma, extracted from standard EDTA-containing blood collection tubes, may contain traces of blood cells consequently resulting in a false low-level HIV-1 viral load when using Roche Cobas HIV-1 assays. Methods: The presence of human DNA in Roche Cobas 4800 RNA extracts and in RNA extracts from the Abbott HIV-1 RealTime assay was assessed by quantifying the human albumin gene by means of quantitative PCR. RNA was extracted from plasma samples before and after an additional centrifugation and tested for viral load and DNA contamination. The relation between total DNA content and viral load was defined. Results: Elevated concentrations of genomic DNA were detected in 28 out of 100 Cobas 4800 extracts and were significantly more frequent in samples processed outside of the AIDS Reference Laboratory. An association between genomic DNA presence and spurious low-level viraemia results was demonstrated. Supplementary centrifugation of plasma before RNA extraction eliminated the contamination and the false viraemia. Conclusions: Plasma isolated from standard EDTA-containing blood collection tubes may contain traces of HIV DNA leading to false viral load results above the clinical cutoff. Supplementary centrifugation of plasma before viral load analysis may eliminate the occurrence of this spurious low-level viraemia.
Keywords
QUANTIFICATION, SAMPLES, RNA, DNA

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MLA
Mortier, Virginie et al. “Meticulous Plasma Isolation Is Essential to Avoid False Low-level Viraemia in Roche Cobas HIV-1 Viral Load Assays.” ANTIVIRAL THERAPY 23.3 (2018): 277–281. Print.
APA
Mortier, V., Vancoillie, L., Dauwe, K., Staelens, D., Demecheleer, E., Schauvliege, M., Dinakis, S., et al. (2018). Meticulous plasma isolation is essential to avoid false low-level viraemia in Roche Cobas HIV-1 viral load assays. ANTIVIRAL THERAPY, 23(3), 277–281.
Chicago author-date
Mortier, Virginie, Leen Vancoillie, Kenny Dauwe, Delfien Staelens, Els Demecheleer, Marlies Schauvliege, Sylvie Dinakis, et al. 2018. “Meticulous Plasma Isolation Is Essential to Avoid False Low-level Viraemia in Roche Cobas HIV-1 Viral Load Assays.” Antiviral Therapy 23 (3): 277–281.
Chicago author-date (all authors)
Mortier, Virginie, Leen Vancoillie, Kenny Dauwe, Delfien Staelens, Els Demecheleer, Marlies Schauvliege, Sylvie Dinakis, Tom Van Maerken, Géraldine Dessilly, Jean Ruelle, and Chris Verhofstede. 2018. “Meticulous Plasma Isolation Is Essential to Avoid False Low-level Viraemia in Roche Cobas HIV-1 Viral Load Assays.” Antiviral Therapy 23 (3): 277–281.
Vancouver
1.
Mortier V, Vancoillie L, Dauwe K, Staelens D, Demecheleer E, Schauvliege M, et al. Meticulous plasma isolation is essential to avoid false low-level viraemia in Roche Cobas HIV-1 viral load assays. ANTIVIRAL THERAPY. 2018;23(3):277–81.
IEEE
[1]
V. Mortier et al., “Meticulous plasma isolation is essential to avoid false low-level viraemia in Roche Cobas HIV-1 viral load assays,” ANTIVIRAL THERAPY, vol. 23, no. 3, pp. 277–281, 2018.
@article{8553023,
  abstract     = {Background: Pre-analytical sample processing is often overlooked as a potential cause of inaccurate assay results. Here we demonstrate how plasma, extracted from standard EDTA-containing blood collection tubes, may contain traces of blood cells consequently resulting in a false low-level HIV-1 viral load when using Roche Cobas HIV-1 assays. 
Methods: The presence of human DNA in Roche Cobas 4800 RNA extracts and in RNA extracts from the Abbott HIV-1 RealTime assay was assessed by quantifying the human albumin gene by means of quantitative PCR. RNA was extracted from plasma samples before and after an additional centrifugation and tested for viral load and DNA contamination. The relation between total DNA content and viral load was defined. 
Results: Elevated concentrations of genomic DNA were detected in 28 out of 100 Cobas 4800 extracts and were significantly more frequent in samples processed outside of the AIDS Reference Laboratory. An association between genomic DNA presence and spurious low-level viraemia results was demonstrated. Supplementary centrifugation of plasma before RNA extraction eliminated the contamination and the false viraemia. 
Conclusions: Plasma isolated from standard EDTA-containing blood collection tubes may contain traces of HIV DNA leading to false viral load results above the clinical cutoff. Supplementary centrifugation of plasma before viral load analysis may eliminate the occurrence of this spurious low-level viraemia.},
  author       = {Mortier, Virginie and Vancoillie, Leen and Dauwe, Kenny and Staelens, Delfien and Demecheleer, Els and Schauvliege, Marlies and Dinakis, Sylvie and Van Maerken, Tom and Dessilly, Géraldine and Ruelle, Jean and Verhofstede, Chris},
  issn         = {1359-6535},
  journal      = {ANTIVIRAL THERAPY},
  keywords     = {QUANTIFICATION,SAMPLES,RNA,DNA},
  language     = {eng},
  number       = {3},
  pages        = {277--281},
  title        = {Meticulous plasma isolation is essential to avoid false low-level viraemia in Roche Cobas HIV-1 viral load assays},
  url          = {http://dx.doi.org/10.3851/imp3203},
  volume       = {23},
  year         = {2018},
}

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