Comparison of three methods for extraction of HCV RNA in sera collected from individuals with hyperlipidemia, hyperbilirubinemia and hyperglobulinemia

Zhang, Baohua; Chen, Kun; Ni, Erru

doi:10.1016/j.jviromet.2014.11.004

Cited by 4 publications

(2 citation statements)

References 16 publications

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“…The impact of RNA or DNA extraction method has been evaluated in several studies quantifying HIV [15], HCV [16] or HBV viral load [17][18][19], with the manual extraction methods having better, equal or worse performance than automated methods in different settings. In a study evaluating HBV and HCV viral load quantification, the authors compared the performance of an automated extraction method (NucliSens easyMAG; BioMérieux, Craponne, France) and a manual extraction method (QIAmpMin Elute Kit; Qiagen, Hilden, Germany).…”

Section: Discussionmentioning

confidence: 99%

Automated Nucleic Acid Isolation Methods for HDV viral Load Quantification can Lead to viral Load Underestimation

et al. 2018

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Background: HDV infection is a cause of severe liver disease. Diagnosis and monitoring of HDV RNA are important to patient management. Since 2012, a WHO standard for HDV RNA quantification has been available; however, the impact of RNA extraction methods on HDV viral load quantification has never been evaluated. Methods: The aim of this study was to compare four commonly used automated nucleic acid (NA) extraction methods (AmpliPrep, MagNA Pure, QIAcube QBK and QIAcube VRK) with a manual RNA extraction method (Instant Virus RNA/DNA kit) and evaluate the possible effect of each method on HDV RNA yield with subsequent amplification with the Robogene HDV assay. Serum samples from HDVpositive patients taken before treatment with pegylated interferon-a2a and at treatment weeks 12 and 48 were studied. Results: The automated extraction methods MagNA Pure, Ampliprep and QIAcube VRK extraction led to about 10-fold lower HDV RNA values compared with the manual method of NA extraction, while the difference was smaller with QIAcube QBK (about 6-fold lower). The median viral load was 10,665 IU/ml for the manual method, 445 IU/ml for AmpliPrep, 3,209 IU/ml for MagNA Pure, 2,060 IU/ml for QIAcube QBK and 3,568 IU/ml for QIAcube VRK. Use of MagNA Pure led to misclassification of two on-treatment samples with low viral load as being false negative. Conclusions: The NA extraction method had a significant impact on the measured HDV viral loads determined by the commonly used Robogene assay, with the manual RNA method yielding consistently higher values of viral load. ClinicalTrials.gov Identifier: NCT00932971.

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Section: Discussionmentioning

confidence: 99%

Automated Nucleic Acid Isolation Methods for HDV viral Load Quantification can Lead to viral Load Underestimation

et al. 2018

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“…Although the introduction of real-time PCR has led to considerable progress in automating the amplification and detection steps, still nucleic acid isolation remains very labor-intensive when performed manually. Thus, the objective of the present study was to evaluate performance of vacuum manifold system and spin based protocols for extraction of HCV-RNA using Favorgen kit high efficiency silica based spin column as reported in [ 4 ] followed by RT-qPCR. The sensitivity of two methods was compared using samples of different viral load.…”

Section: Introductionmentioning

confidence: 99%

Extraction of HCV-RNA from Plasma Samples: Development towards Semiautomation

Amin¹,

Jabbar²,

Niazi³

et al. 2015

International Journal of Analytical Chemistry

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A semiautomated extraction protocol of HCV-RNA using Favorgen RNA extraction kit has been developed. The kit provided protocol was modified by replacing manual spin steps with vacuum filtration. The assay performance was evaluated by real-time qPCR based on Taqman technology. Assay linearity was confirmed with the serial dilutions of RTA (Turkey) containing 1 × (106, 105, 104, and 103) IU mL−1. Comparison of test results obtained by two extraction methods showed a good correlation (r = 0.95, n = 30) with detection limit of 102 IU mL−1. The semiautomated vacuum filtration based protocol demonstrated high throughput: 35 minutes for the extraction of a batch of 30 samples (150 µL each) with reduced labor, time, waste, and cost. Performance characteristics of semiautomated system make it suitable for use in diagnostic purpose and viral load determinations.

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Detection of arboviruses in blood and mosquito slurry samples using polymer microchip

Prakash

Wong²,

Tellier

et al. 2017

2017 IEEE Healthcare Innovations and Point of Care Technologies (HI-POCT)

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Comparison of three methods for extraction of HCV RNA in sera collected from individuals with hyperlipidemia, hyperbilirubinemia and hyperglobulinemia

Cited by 4 publications

References 16 publications

Automated Nucleic Acid Isolation Methods for HDV viral Load Quantification can Lead to viral Load Underestimation

Automated Nucleic Acid Isolation Methods for HDV viral Load Quantification can Lead to viral Load Underestimation

Extraction of HCV-RNA from Plasma Samples: Development towards Semiautomation

Detection of arboviruses in blood and mosquito slurry samples using polymer microchip

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