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Comparative Study
. 2009 Aug;47(8):2635-8.
doi: 10.1128/JCM.02170-08. Epub 2009 Jun 17.

Detection and quantification of hepatitis C virus (HCV) by MultiCode-RTx real-time PCR targeting the HCV 3' untranslated region

Elizabeth K Mulligan  1 , J J Germer, Max Q Arens, Krista L D'Amore, Adrian Di Bisceglie, Nathan A Ledeboer, Michael J Moser, Andrew C Newman, Andy K O'Guin, Paul D Olivo, Diane S Podzorski, Kimberly A Vaughan, Joseph D Yao, Slava A Elagin, Scott C Johnson
Affiliations
Comparative Study

Detection and quantification of hepatitis C virus (HCV) by MultiCode-RTx real-time PCR targeting the HCV 3' untranslated region

Elizabeth K Mulligan et al. J Clin Microbiol. 2009 Aug.

Abstract

A prototype, real-time reverse-transcription PCR assay, based on MultiCode-RTx technology, quantifying hepatitis C virus (HCV) RNA by targeting the HCV 3' untranslated region demonstrated linearity over 7 logs, with a good correlation between the quantitative results of this assay and the results of two commercially available comparator assays for 466 clinical specimens comprising all six HCV genotypes.

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Figures

FIG. 1.
FIG. 1.
Linearity of the RTx assay. (A) The upper graph shows real-time PCR fluorescence data as baseline-adjusted relative fluorescence units (RFU) versus the number of PCR cycles. The lower graph shows Tm analysis of the negative value of the change in RFU over the change in temperature (−dRFU/dT) versus temperature (degrees Celsius). (B) Linear relationship between RTx assay CT values and 3′ UTR target copy numbers/reaction. Eff, efficiency.
FIG. 2.
FIG. 2.
Differences in HCV VL results with a worldwide HCV performance panel of six HCV genotypes tested by three assay methods (•, small-volume RTx assay result − bDNA assay result; □, small-volume RTx assay result − CAM test result). The dotted line indicates the mean difference between small-volume RTx assay and bDNA assay results, and the dashed line indicates the mean difference between small-volume RTx assay and CAM test results.
FIG. 3.
FIG. 3.
Comparisons of small-volume RTx assay results to bDNA and TaqMan results for clinical serum and plasma specimens for quantification of HCV RNA. (A) Linear regression analysis of the small-volume RTx assay versus the bDNA assay for 273 specimens. (B) Bland-Altman plot of paired VL differences between the small-volume RTx assay and the bDNA assay. (C) Linear regression analysis of the small-volume RTx assay versus the TaqMan assay for 98 specimens. (D) Bland-Altman plot of paired VL differences between the small-volume RTx and TaqMan assay VLs. HCV genotypes in panels B and D are indicated by the following symbols: •, genotype 1; ▵, genotype 2; ○, genotype 3; ×, genotype 4; ▴, genotype 5; ▪, genotype 6; □, genotype unspecified.
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