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Vol. 14. Issue 2.
Pages 147-152 (March - April 2010)
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Vol. 14. Issue 2.
Pages 147-152 (March - April 2010)
Original article
Open Access
Signal to cut-off (S/CO) ratio and detection of HCV genotype 1 by real-time PCR one-step method: is there any direct relationship?
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Guilherme Albertoni
Corresponding author
albertonig@nefro.epm.br

Correspondence to: Av. Jandira, 1260, Indianópolis São Paulo – SP – Brazil CEP: 04080-006. Phone: 55 11 50556588/R. 39 Fax: 55 11 50556588.
, C.P. Arnoni, P.R.B. Araújo, F.O. Carvalho, J.A. Barreto
Colsan – Associação Beneficente de Coleta de Sangue, São Paulo, SP, Brazil
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Abstract
Background

Polymerase chain reaction (PCR) methods play an essential role in providing data related to diagnosis, monitoring and treatment of hepatitis C virus (HCV) infection. EIA results are reported as “reactive” or “non reactive” and EIA S/CO ratio may also be reported as “high” or “low.” This study aimed to evaluate the performance of a real-time RT-PCR and assess whether there is relationship between S/CO and PCR results.

Study Design and Methods

Sera from blood donors were analyzed by Enzyme-Linked Immunosorbent Assay (ELISA) and RT-PCR assay to detect HCV infection.

Results

The RT-PCR assay to genotypes 1a/b showed an acceptable linear response in serial dilutions. The samples were divided into two groups based on their serological results: group A – S/CO ratio < 3 (60 samples) and group B – S/CO ratio > 3 (41 samples). Viral loads were confirmed positive in group B samples in 90%, and in group A samples were confirmed positive in only 13% by RT-PCR.

Conclusion

The methodology used was able to detect the presence of RNA-HCV genotype I in 90% of the samples serologically positive in group B. All negative samples were sent to search for other genotypes of HCV (genotypes 2-6) and were confirmed as negative. These data suggests that these negative samples may have HCV RNA viral load below the detection limit of our test (310 IU/ mL), or a false positive result in serological test, or spontaneous viral clearance occurred.

Keywords:
HCV
HCV genotype 1
real-time PCR
ELISA
RNA extraction
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