A modified competitive RT-PCR (mcRT-PCR) to measure HCV RNA in serum and the Amplicor HCV Monitor assay were compared. For mcRT-PCR, the RNA extracted was retrotranscribed and coamplified in one step with a known amount of a DNA internal control (IC). Digoxigenin-labeled amplified products were hybr
Performance of two Real-Time RT-PCR assays for quantitation of hepatitis C virus RNA: Evaluation on HCV genotypes 1–4
✍ Scribed by Abeer Elkady; Yasuhito Tanaka; Fuat Kurbanov; Fuminaka Sugauchi; Masaya Sugiyama; Anis Khan; Elsayed Mostafa Ali; Layla Mouhamed; Sahar Abou el-fetouh; AbdEl-Rahaman AbdEl-Hameed; Masashi Mizokami
- Publisher
- John Wiley and Sons
- Year
- 2010
- Tongue
- English
- Weight
- 583 KB
- Volume
- 82
- Category
- Article
- ISSN
- 0146-6615
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✦ Synopsis
Abstract
Accuracy for monitoring of the concentration of hepatitis C virus (HCV) RNA represents a major challenge throughout the management of patients with chronic hepatitis C. To investigate the genotype‐independent efficiency and the accuracy of two real‐time detection reverse transcription‐polymerase chain reaction (RT‐PCR) assays; the Cobas Ampliprep/Cobas TaqMan (CAP/CTM); and the Abbott RealTime HCV (ART), a total of 184 samples with different HCV subtypes were examined; 1b (n = 58), 2a (n = 39), 2b (n = 26), 3a (n = 20), and 4 (n = 41). A robust linear correlation was observed between the two assays applied to genotypes 1b, 2a, 2b, and 3a [the correlation coefficient (R) ranged from 0.99 to 0.98], but not to genotype 4 specimens (R = 0.78). A significant difference in measurements of HCV RNA using CAP/CTM and ART in serum samples with genotypes 1b and 4 was observed (0.72, −0.53 log IU/ml, P < 0.0001, 0.01, respectively). A robust correlation was observed between the HCV core antigen and HCV RNA values by either of the HCV RNA quantitation assays applied to all genotypes with exception of genotype 4, for which R was higher with ART (R = 0.95) than with CAP/CTM (R = 0.80). The lower limit of detection of CAP/CTM and ART were 41.4 and 28.5 IU/ml using the WHO standards, respectively. In conclusion, two RT‐PCR assays had a high efficiency and accuracy for quantitation of HCV RNA of genotypes 2a, 2b, and 3a, but the mean values of HCV RNA differed for genotype 1b and 4. J. Med. Virol. 82:1878–1888, 2010. © 2010 Wiley‐Liss, Inc.
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