Hepatitis C virus RNA quantitation in venous and capillary small-volume whole-blood samples.
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-microl sample (95% limit of detection [95% LOD]), a linearity range of 500 to 5 x 10(6) 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-microl 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-microl WB samples is appropriate for monitoring viral loads of >900 IU/ml, although the use of WB does not increase the diagnostic sensitivity.
Bruns
Tony
T
Division of Gastroenterology, Hepatology and Infectious Disease, Department of Internal Medicine II, Friedrich Schiller University, Jena University Hospital, Jena, Germany. tony.bruns@med.uni-jena.de
Steinmetzer
Katrin
K
Ermantraut
Eugen
E
Stallmach
Andreas
A
D001769:Blood / Q000821:virology; D002196:Capillaries / Q000821:virology; D016174:Hepacivirus / Q000302:isolation & purification*; D019698:Hepatitis C, Chronic / Q000821:virology; D006801:Humans; D012367:RNA, Viral / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D019562:Viral Load / Q000379:methods*
D012367:RNA, Viral
Lancet. 1986 Feb 8;1(8476):307-10
2868172
Proc Natl Acad Sci U S A. 1997 Aug 5;94(16):8738-43
9238047
J Viral Hepat. 1996 Nov;3(6):285-92
8947879
Nucleic Acids Res. 2004 Apr 19;32(7):e64
15096560
J Virol. 2007 Jul;81(14):7710-5
17475654
Hepatology. 2005 Jan;41(1):106-14
15619235
Lancet. 2002 Jun 29;359(9325):2230-4
12103286
J Gen Virol. 2006 Aug;87(Pt 8):2243-2251
16847120
J Clin Virol. 2002 Aug;25(2):205-16
12367656
Liver Int. 2009 Jan;29 Suppl 1:74-81
19207969
Bioorg Med Chem Lett. 1998 Aug 18;8(16):2219-22
9873516
J Hepatol. 2008 Jun;48(6):932-8
18433918
J Clin Microbiol. 1999 Mar;37(3):484-9
9986800
World J Gastroenterol. 2001 Apr;7(2):228-34
11819765
J Clin Virol. 2007 Apr;38(4):326-33
17344093
Hepatobiliary Pancreat Dis Int. 2005 Nov;4(4):550-3
16286260
J Infect Dis. 1992 Dec;166(6):1276-80
1385547
J Clin Virol. 2004 Dec;31(4):312-3
15494276
NIH Consens State Sci Statements. 2002 Jun 10-12;19(3):1-46
14768714
Clin Infect Dis. 2004 Dec 15;39(12):1754-60
15578395
J Clin Virol. 2007 Aug;39(4):308-11
17625963
Hepatology. 2007 Jul;46(1):22-31
17525931
Blood. 1994 Jan 1;83(1):269-73
8274741
J Clin Microbiol. 2008 Nov;46(11):3791-4
18701664
J Clin Microbiol. 2007 Jul;45(7):2151-5
17494720
Blood. 2001 Oct 15;98(8):2293-300
11588022
Nucleic Acids Res. 1991 May 25;19(10):2629-35
1710340
J Infect Dis. 1997 Nov;176(5):1209-14
9359720
J Infect Dis. 1997 Jul;176(1):20-6
9207345
Scand J Gastroenterol. 2002 May;37(5):578-84
12059061
J Infect Dis. 2004 Jan 1;189(1):7-14
14702147
J Virol. 1999 Nov;73(11):9213-21
10516029
J Clin Microbiol. 2000 Dec;38(12):4326-31
11101559
J Hepatol. 2005 Jan;42(1):150-2
15629525
Vox Sang. 1999;76(3):149-58
10341329
Clin Chem. 2007 Nov;53(11):1962-5
17761752
Blood. 1996 Oct 1;88(7):2768-74
8839874
Dig Liver Dis. 2007 Sep;39 Suppl 1:S38-45
17936221
Hepatogastroenterology. 2003 Sep-Oct;50(53):1301-4
14571723
J Clin Microbiol. 2006 Mar;44(3):729-37
16517847
J Clin Chem Clin Biochem. 1983 Nov;21(11):709-20
6655447
Bioinformatics. 2005 Feb 1;21(3):379-84
15377502