Abstract
Background: Accurate viral load (VL) determination is paramount to determine the efficacy of anti-HIV-1 therapy. The conventional method used, fit-point (FP), assumes an equal efficiency in the polymerase chain reaction (PCR) among samples that might not hold for low-input templates. An alternative approach, maxRatio, was introduced to compensate for inhibition in PCR. Methods: Herein, we assessed whether maxRatio could improve VL quantification using 2,544 QIAgen artus HI virus-1 RT-PCR reactions. The assay's standard dilutions were used to build external standard curves with either FP or maxRatio that re-calculated the VLs. Results: FP and maxRatio were highly comparable (Pearson's ρ=0.994, Cohen's κ=0.885), and the combination of the two methods identified samples (n=41) with aberrant amplification profiles. Conclusions: The combination of maxRatio and FP could improve the predictive value of the assay.
Highlights
Infection with HIV-1 accounts for a global prevalence of 38 million cases and a one million deaths yearly[1]
We compared the quantification of HIV-1 viral load (VL) computed by FP and maxRatio on a dataset generated with the QIAgen artus HIV assay, which has a reported limit of detection of 35.5 copies per milliliter (c/mL), and we showed that maxRatio could pinpoint samples with abnormal amplification profiles
The VLs obtained with the two methods were very strongly correlated (ρ = 0.994, 95% confidence interval (CI): 0.993-0.994) and the agreement in the stratification of the reactions into reactive and non-reactive was almost perfect (κ = 0.885, 95% CI: 0.863-0.907)
Summary
Infection with HIV-1 accounts for a global prevalence of 38 million cases and a one million deaths yearly[1]. An accurate viral load (VL), typically carried out by quantitative polymerase chain reaction (qPCR), is pivotal for addressing the efficacy of antiviral therapies[2]. QPCR data are usually analyzed by the fit-point (FP) method, which assumes equal amplification efficiency between samples[5]. MaxRatio, was introduced to overcome these issues[9]. Accurate viral load (VL) determination is paramount to determine the efficacy of anti-HIV-1 therapy. The conventional method used, fit-point (FP), assumes an equal efficiency in the polymerase chain reaction (PCR) among samples that might not hold for low-input templates. MaxRatio, was introduced to compensate for inhibition in PCR. Methods: we assessed whether maxRatio could improve VL quantification using 2,544 QIAgen artus HI virus-1 RT-PCR reactions. Conclusions: The combination of maxRatio and FP could improve the predictive value of the assay
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
