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Amplifying and Quantifying HIV-1 RNA in HIV Infected Individuals with Viral Loads Below the Limit of Detection by Standard Clinical Assays

Amplifying and Quantifying HIV-1 RNA in HIV Infected Individuals with Viral Loads Below the Limit of Detection by Standard Clinical Assays

Published on: September 26, 2011

Early infant diagnosis HIV-1 PCR cycle-threshold predicts infant viral load at birth

Ahmad Haeri Mazanderani¹, Tendesayi Kufa², Karl G Technau³

¹Centre for HIV and STIs, National Institute for Communicable Diseases, National Health Laboratory Service, Johannesburg, South Africa; Department of Medical Virology, Faculty of Health Sciences, University of Pretoria, Pretoria, South Africa.

Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology

|March 24, 2019

Insights

Qualitative polymerase chain reaction (PCR) cycle-threshold (Ct) values from early infant diagnosis (EID) tests at birth can predict human immunodeficiency virus type 1 (HIV-1) viral load (VL). This allows for early identification of infants with high VL, crucial for timely intervention.

Area of Science:

Pediatric Infectious Diseases
Virology
Molecular Diagnostics

Background:

HIV-1 viral load (VL) is a key predictor of disease progression in untreated children.
Current early infant diagnosis (EID) primarily uses qualitative polymerase chain reaction (PCR) assays.

Purpose of the Study:

To determine if qualitative EID real-time PCR cycle-threshold (Ct) values obtained at birth can predict HIV-1 VL.
To identify specific Ct value thresholds for predicting high VL in neonates.

Main Methods:

Secondary analysis of intrauterine HIV-1 infected neonates (n=107) in Johannesburg, South Africa.
Utilized EID HIV-1 PCR testing (COBAS AmpliPrep/COBAS TaqMan and Xpert HIV-1 Qualitative assays) and plasma HIV-1 RNA VL testing.
Employed Bland-Altman analysis and multivariable linear regression to assess agreement and predict VL based on Ct values.

Main Results:

Good correlation (Spearman r=0.9) observed between Ct values of the two EID assays.
Each one-cycle increase in Ct value corresponded to a 0.3 log10 decrease in RNA.
EID CAP/CTM Ct ≤ 23 and EID Xpert Ct ≤ 31 accurately predicted VL > 5.0 log10 cps/ml in over 82% of cases.

Conclusions:

Qualitative EID PCR Ct values at birth are valuable predictors of HIV-1 VL in neonates.
Specific Ct value thresholds can reliably identify infants with high viral loads, enabling prompt clinical management.

Read full analysis

Abstract

Background:

HIV-1 viral load (VL) has been found to be an independent predictor for disease progression among untreated HIV-infected children. However, qualitative polymerase chain reaction (PCR) assays are routinely used for early infant diagnosis (EID).

Objectives:

To predict HIV-1 VL at birth using qualitative EID real-time PCR cycle-threshold (Ct) values.

Study Design:

This study was a secondary analysis of results from a cohort of intrauterine HIV-1 infected neonates. Neonates were enrolled at Rahima Moosa Mother & Child Hospital in Johannesburg, South Africa between June 2014 and November 2017. Laboratory EID HIV-1 PCR testing was performed at birth using COBAS AmpliPrep/COBAS TaqMan HIV-1 Qualitative Test v2.0 (EID CAP/CTM). Some infants had simultaneous EID point-of-care (POC) testing using Xpert HIV-1 Qualitative assay (EID Xpert). Neonates with a confirmed HIV-1 detected EID result and plasma HIV-1 RNA VL test were included in this analysis. Bland-Altman analysis was used to determine extent of agreement between Ct values of both EID assays. Multivariable linear regression models adjusting for time between EID and VL testing were used to describe the association between EID Ct values and VL and to predict VL at given EID Ct values.

Results:

Among 107 HIV-1 infected neonates included in the study, 59 had POC EID testing. Median VL was 28 400 copies per millilitre (cps/ml) (IQR: 1 918-218 358) - two neonates had VL < 100 cps/ml prior to antiretroviral therapy initiation. There was good correlation between Ct values of both EID assays (Spearman correlation coefficient 0.9, 95% CI: 0.8-1.0). The limits of agreement between EID CAP/CTM and Xpert Ct values were 4-11 cycles. For every one cycle increase in Ct value there was 0.3 log₁₀ RNA decrease (95% CI: -0.3 to -0.2) for both EID assays. An EID CAP/CTM Ct value ≤ 23 and an EID Xpert Ct value ≤ 31 predicted a VL of > 5.0 log₁₀ cps/ml in 82.2% (95% CI: 73.9-88.3) and 84.7% (95% CI: 73.7-91.8) of cases, respectively.

Conclusion:

EID Ct values at birth predict VL and accurately identify infants with VL > 5.0 log₁₀ cps/ml.

Keywords:

Early infant diagnosis HIV Point of care Viral load

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