Comparative study of a modified competitive RT-PCR and amplicor HCV monitor assays for quantitation of hepatitis C virus RNA in serum
| العنوان: | Comparative study of a modified competitive RT-PCR and amplicor HCV monitor assays for quantitation of hepatitis C virus RNA in serum |
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| المؤلفون: | Xavier Forns, Josep Costa, M Guilera, Joan Rodés, M. D. Maluenda, E Olmedo, Sergi Ampurdanés, F X López-Labrador, José-María Sánchez-Tapias, M. T. Jimenez de Anta |
| المصدر: | Journal of Medical Virology. 58:35-43 |
| بيانات النشر: | Wiley, 1999. |
| سنة النشر: | 1999 |
| مصطلحات موضوعية: | biology, Hepatitis C virus, Hepacivirus, virus diseases, Alpha interferon, RNA, medicine.disease_cause, biology.organism_classification, Virology, law.invention, Flaviviridae, Infectious Diseases, Real-time polymerase chain reaction, law, medicine, Viral load, Polymerase chain reaction |
| الوصف: | 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 hybridized to specific HCV DNA and IC-DNA probes and quantified by colorimetry. HCV RNA concentration was calculated by plotting the ratio of HCV/IC ODs against a calibration curve. Multiple samples were analyzed in the same round and tedious titration of each sample with a competitor was unnecessary. The mcRT-PCR assay was linear from 6 × 103 to 6 × 107 copies/ml, whereas Amplicor was linear up to 1–2 × 106 copies/ml. HCV RNA was measured in samples from 75 carriers. There was agreement between both methods in type 1 infections but not in type 2 or type 3 infections, in which the values measured by Amplicor were, on average, 15 times lower than those measured by the mcRT-PCR. HCV RNA measured by Amplicor was higher in type 1 infections than in type 2 or 3 infections, but no differences were found when viral load was assessed by mcRT-PCR. The binding efficiency of the Amplicor-probe was greater for type 1 than for types 2 or 3, suggesting Amplicor underestimates the viral load in the latter types. In contrast, the mcRT-PCR is not affected by genotype-related variation of HCV. This study suggests that mcRT-PCR assay is reliable for sensitive and accurate measurement of HCV RNA over a broad range of values independently of the HCV genotype. J. Med. Virol. 58:35–43, 1999. © 1999 Wiley-Liss, Inc. |
| تدمد: | 1096-9071 0146-6615 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_________::31bb8de6b68a271a3e67b70c94876dbb https://doi.org/10.1002/(sici)1096-9071(199905)58:1<35::aid-jmv5>3.0.co;2-v |
| حقوق: | CLOSED |
| رقم الأكسشن: | edsair.doi...........31bb8de6b68a271a3e67b70c94876dbb |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10969071 01466615 |
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