Development of cost-effective real-time PCR test: to detect a wide range of HBV DNA concentrations in the western Amazon region of Brazil.
Virol J
; 11: 16, 2014 Jan 28.
Article
em En
| MEDLINE
| ID: mdl-24472141
ABSTRACT
BACKGROUND:
Currently there is a significant risk of infection with hepatitis B virus (HBV) during blood transfusion in high epidemic area. This is due to the pre-seroconversion window period, immunovariant viral strains and the presence of occult HBV infection (OBI). The aim of this study was to develop an in-house real-time PCR-based method, which was both ultra-sensitive and efficient offering an alternative method for nucleic acid testing (NAT).METHODS:
A precore fragment with 109 bp was cloned and serial diluted to standard curve construction. The calibration of the HBV-DNA values was performed against OptiQuant® HBV-DNA Quantification Panel, Acrometrix Europe B.V.).RESULTS:
From our in-house plasmid we prepared serial dilutions ranging from 2 × 10³-2 × 109 copies/ml. The threshold was adjusted automatically during analysis and the data collected were analyzed by linear regression (r² = 0.99). The limit of detection for the assay with pHBVRO standards was 2000/ml in a total reaction volume of 30 µl. We found a strong correlation between the two methods (r² = 0.9965 and p < 0.0001). The regression line give us the following equation Log 10 (IU/mL) = 0.9038Log 10 (copies/mL)--1.0643, suggesting that 1 IU/mL = 15 copies/mL.CONCLUSIONS:
Therefore, we can affirm that the qHBVRO PCR can detect HBV DNA in individuals with hepatitis B at any stage of the disease showing high capacity for NAT screening in hepatitis b donors. This results of sensitivity could provide an advance for automation in blood banks and increasing safety of patients who receive blood transfusions.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Vírus da Hepatite B
/
Carga Viral
/
Reação em Cadeia da Polimerase em Tempo Real
/
Hepatite B
Tipo de estudo:
Diagnostic_studies
/
Evaluation_studies
/
Health_economic_evaluation
Limite:
Humans
País/Região como assunto:
America do sul
/
Brasil
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article