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Journal of Clinical Microbiology, October 2009, p. 3231-3240, Vol. 47, No. 10
0095-1137/09/$08.00+0     doi:10.1128/JCM.00925-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Hepatitis C Virus RNA Quantitation in Venous and Capillary Small-Volume Whole-Blood Samples{triangledown}

Tony Bruns,1,2* Katrin Steinmetzer,2 Eugen Ermantraut,2 and Andreas Stallmach1

Division of Gastroenterology, Hepatology and Infectious Disease, Department of Internal Medicine II, Jena University Hospital, Jena, Germany,1 Clondiag GmbH, Jena, Germany2

Received 12 May 2009/ Returned for modification 8 July 2009/ Accepted 11 August 2009

Quantitation of hepatitis C virus (HCV) RNA in plasma and serum samples is a costly procedure in both time and reagents. Additionally, cell-associated viral RNA may not be detected. This study evaluated the accuracy of HCV RNA quantitation in small-volume whole-blood (WB) samples, which would be appropriate for point-of-care diagnostic devices. HCV RNA was extracted from 222 clinical plasma and WB samples of 82 patients with chronic hepatitis C by a specific locked nucleic acid-mediated capture method and quantified by real-time reverse transcription-PCR. The results were compared to the reference plasma viral load determined with the COBAS AmpliPrep/TaqMan (CAP/CTM) HCV test. This assay had an analytical sensitivity of 9 IU per 10-µl sample (95% limit of detection [95% LOD]), a linearity range of 500 to 5 x 106 IU/ml, and was accurate in testing 10 HCV subtypes (<0.22 log10 unit) in plasma. The assay was matrix equivalent for plasma and WB samples (coefficient of determination [R2] of 0.943) and had a specificity of 100% (n = 20) in WB samples. The HCV RNA concentration in clinical WB samples exceeded the estimated hematocrit-corrected plasma viral loads by 0.22 log10 unit, but absolute quantitation results in plasma and WB samples were identical (95% confidence interval, –0.06 to 0.04 log10 unit). The sensitivity in WB samples was 100% (n = 141) for plasma concentrations above the 95% LOD. Quantitation results in 10-µl WB samples correlated linearly with the CAP/CTM HCV plasma test results (R2 = 0.919; n = 140) and did not differ between capillary and venous samples (R2 = 0.960; n = 40). This study shows that HCV RNA quantitation in 10-µl WB samples is appropriate for monitoring viral loads of >900 IU/ml, although the use of WB does not increase the diagnostic sensitivity.

* Corresponding author. Mailing address: Department of Gastroenterology, Hepatology and Infectious Disease, Clinic of Internal Medicine II, Friedrich Schiller University, Erlanger Allee 101, 07740 Jena, Germany. Phone: 49 3641 9324585. Fax: 49 3641 9324222. E-mail: tony.bruns{at}med.uni-jena.de

{triangledown} Published ahead of print on 19 August 2009.

Journal of Clinical Microbiology, October 2009, p. 3231-3240, Vol. 47, No. 10
0095-1137/09/$08.00+0     doi:10.1128/JCM.00925-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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