Ann Lab Med.  2017 Mar;37(2):147-150. 10.3343/alm.2017.37.2.147.

Performance of the Real-Q EBV Quantification Kit for Epstein-Barr Virus DNA Quantification in Whole Blood

• Huh HJ
• Park JE
• Kim JY
• Yun SA
• Lee MK
• Lee NY
• Kim JW
• Ki CS

• KMID: 2373632
• DOI: http://doi.org/10.3343/alm.2017.37.2.147

Abstract

There has been increasing interest in standardized and quantitative Epstein-Barr virus (EBV) DNA testing for the management of EBV disease. We evaluated the performance of the Real-Q EBV Quantification Kit (BioSewoom, Korea) in whole blood (WB). Nucleic acid extraction and real-time PCR were performed by using the MagNA Pure 96 (Roche Diagnostics, Germany) and 7500 Fast real-time PCR system (Applied Biosystems, USA), respectively. Assay sensitivity, linearity, and conversion factor were determined by using the World Health Organization international standard diluted in EBV-negative WB. We used 81 WB clinical specimens to compare performance of the Real-Q EBV Quantification Kit and artus EBV RG PCR Kit (Qiagen, Germany). The limit of detection (LOD) and limit of quantification (LOQ) for the Real-Q kit were 453 and 750 IU/mL, respectively. The conversion factor from EBV genomic copies to IU was 0.62. The linear range of the assay was from 750 to 10⁶ IU/mL. Viral load values measured with the Real-Q assay were on average 0.54 log₁₀ copies/mL higher than those measured with the artus assay. The Real-Q assay offered good analytical performance for EBV DNA quantification in WB.

Keyword

Epstein-Barr virus; Real-time PCR; Quantification; Whole blood; Performance

MeSH Terms

DNA, Viral/*blood/metabolism
Epstein-Barr Virus Infections/diagnosis/virology
Herpesvirus 4, Human/*genetics/isolation & purification
Humans
Limit of Detection
Reagent Kits, Diagnostic
Real-Time Polymerase Chain Reaction
DNA, Viral
Reagent Kits, Diagnostic

Figure

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  Fig. 1 Linearity of the Real-Q EBV Quantification Kit.

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  Fig. 2 Comparison of clinical specimens tested with the Real-Q and artus assays. A total of 38 specimens with EBV DNA load above the limit of quantification in both assays were analyzed. (A) A Bland-Altman plot comparing the artus and Real-Q assays. The solid line represents the mean of difference between copy number values obtained by the Real-Q and the artus assays; dashed lines indicate the 95% confidence interval of the mean of difference. (B) The Passing-Bablok regression analysis. The regression line (solid line) and the 95% confidence interval (dashed lines) are displayed.Abbreviation: EBV, Epstein-Barr virus.

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