2012
DOI: 10.1099/jmm.0.040238-0
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Abstract: The objective of the present study was to evaluate four commercial DNA extraction methods and three PCR protocols for hepatitis B virus (HBV) detection in artificially contaminated oral fluid samples. The extraction protocols were selected based on ease of use and cost, and were also compared with respect to sensitivity and cost. Prior PCR optimization was conducted, in which the sample volume for DNA extraction and the concentrations of DNA and Taq DNA polymerase in the PCR were adjusted. One-round PCR, used … Show more

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Cited by 16 publications
(15 citation statements)
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References 32 publications
(39 reference statements)
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“…Standardization and improvement in the quality and quantity of DNA must be performed for each sample. In this context, Portilho et al observed that laboratories should standardize their own procedures according to the routines of collecting and processing of samples (27) . With this objective, in the present study a previous PCR was conducted to increase the quality and quantity of the viral DNA, both for the sequencing reaction and for the standardization of the multiplex qPCR to study HBV genotypes.…”
Section: Discussionmentioning
confidence: 99%
See 1 more Smart Citation
“…Standardization and improvement in the quality and quantity of DNA must be performed for each sample. In this context, Portilho et al observed that laboratories should standardize their own procedures according to the routines of collecting and processing of samples (27) . With this objective, in the present study a previous PCR was conducted to increase the quality and quantity of the viral DNA, both for the sequencing reaction and for the standardization of the multiplex qPCR to study HBV genotypes.…”
Section: Discussionmentioning
confidence: 99%
“…With a qPCR method that allows genotyping, sequencing and analysis of sequences for diagnostics and treatment are not necessary (35) . The molecular diagnosis of HBV can be accomplished with serum, plasma, or even oral fluid samples (27) . Standardization and improvement in the quality and quantity of DNA must be performed for each sample.…”
Section: Discussionmentioning
confidence: 99%
“…The DNA extraction method chosen for these experiments utilized the RTP ® DNA/RNA Virus Mini kit (Stratec, Germany) because it had previously been evaluated by our group using oral samples (Portilho et al., ) and had good sensitivity when 400 μl of oral fluid was used. According to Mallory et al.…”
Section: Discussionmentioning
confidence: 99%
“…Some studies have shown that HBV DNA can be detected or quantified in oral fluid samples (Heiberg, Hoegh, Ladelund, Niesters, & Hogh, 2010;Kidd-Ljunggren et al, 2006;Portilho, Martins, Lampe, & Villar, 2012;Van Der Ejik et al, 2005), but different collection devices and detection methods were used in the numerous studies, which hinders the comparison of the results. Some studies demonstrated that HBV DNA could be detected more efficiently in non-stimulated whole saliva or using collection devices that employ mechanical friction (Heiberg et al, 2010;Kidd-Ljunggren et al, 2006).…”
Section: Introductionmentioning
confidence: 99%
“…HBsAg and isolated anti-HBc reactive samples were submitted to seminested PCR testing for HBV DNA detection [18] to identify active HBV infection. Anti-HCV reactive samples were submitted to nested PCR testing, and the samples that contained HCV RNA were also genotyped by RFLP [19, 20].…”
Section: Methodsmentioning
confidence: 99%