Imperial College London
Alternate

ProfessorJillGilmour

Faculty of Medicine, Department of Infectious Disease

Honorary Principal Research Fellow 
 
 
 
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Contact

 

+44 (0)20 3315 5098j.gilmour

 
 
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Location

 

J.2.3 Immunology DepartmentChelsea and Westminster HospitalChelsea and Westminster Campus

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Summary

 

Publications

Citation

BibTex format

@article{Kibirige:2022:10.1038/s41598-021-03016-1,
author = {Kibirige, C and Manak, M and King, D and Abel, B and Hack, H and Wooding, D and Liu, Y and Fernandez, N and Dalel, J and Steve, K and Imami, N and Jagodzinski, L and Gilmour, J},
doi = {10.1038/s41598-021-03016-1},
journal = {Scientific Reports},
title = {Development of a Sensitive, Quantitative Assay with Broad Subtype Specificity for Detection of Total HIV-1 Nucleic Acids in Plasma and PBMC},
url = {http://dx.doi.org/10.1038/s41598-021-03016-1},
volume = {12},
year = {2022}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - An LTR-based Quantitative PCR (qPCR) assay was modified and optimized for the quantification of total HIV-1 nucleic acids in plasma and PBMC. TaqMan qPCR primers and probes were designed against the NCBI/LANL HIV-1 compendium database by analyzing sequences used in assays for sensitive cross-clade detection of HIV-1 as reported in the literature and elucidating regions of improved cross-subtype specificity.  Inosine and mixed nucleotide bases were included at polymorphic sites.  Real-time RT-qPCR and qPCR were performed on plasma viral RNA and cellular lysates.  A step-up amplification approach to allow binding of primers across polymorphic regions showed improved sensitivity compared to universal cycling.  Unlike a lead competing laboratory-developed assay, all major HIV-1 subtypes, and a wide range of recombinants from a 127-member diversity panel were detected and accurately quantified in spiked plasmas.  Semi-nested PCR increased detection sensitivity even further.  The assay was able to detect down to 88 copies/mL of HIV-1 in plasma with 95% efficiency or the equivalent of a single infected cell.  The PCR assay will be valuable in studies that monitor very low viral levels including residual or break through HIV-1 in patients receiving antiretroviral therapy, in HIV-1 cure, and in other research studies.
AU  - Kibirige,C
AU  - Manak,M
AU  - King,D
AU  - Abel,B
AU  - Hack,H
AU  - Wooding,D
AU  - Liu,Y
AU  - Fernandez,N
AU  - Dalel,J
AU  - Steve,K
AU  - Imami,N
AU  - Jagodzinski,L
AU  - Gilmour,J
DO  - 10.1038/s41598-021-03016-1
PY  - 2022///
SN  - 2045-2322
TI  - Development of a Sensitive, Quantitative Assay with Broad Subtype Specificity for Detection of Total HIV-1 Nucleic Acids in Plasma and PBMC
T2  - Scientific Reports
UR  - http://dx.doi.org/10.1038/s41598-021-03016-1
UR  - https://www.nature.com/articles/s41598-021-03016-1
UR  - http://hdl.handle.net/10044/1/93881
VL  - 12
ER  -
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