The DOI matches its Crossref record; 4 of the 4 references it cites were found in a registry or an index.
Highly Precise Measurement of HIV DNA by Droplet Digital PCR
Open accessAbstract
Deoxyribonucleic acid (DNA) of the human immunodeficiency virus (HIV) provides the most sensitive measurement of residual infection in patients on effective combination antiretroviral therapy (cART). Droplet digital PCR (ddPCR) has recently been shown to provide highly accurate quantification of DNA copy number, but its application to quantification of HIV DNA, or other equally rare targets, has not been reported. This paper demonstrates and analyzes the application of ddPCR to measure the frequency of total HIV DNA (pol copies per million cells), and episomal 2-LTR (long terminal repeat) circles in cells isolated from infected patients. Analysis of over 300 clinical samples, including over 150 clinical samples assayed in triplicate by ddPCR and by real-time PCR (qPCR), demonstrates a significant increase in precision, with an average 5-fold decrease in the coefficient of variation of pol copy numbers and a >20-fold accuracy improvement for 2-LTR circles. Additional benefits of the ddPCR assay over qPCR include absolute quantification without reliance on an external standard and relative insensitivity to mismatches in primer and probe sequences. These features make digital PCR an attractive alternative for measurement of HIV DNA in clinical specimens. The improved sensitivity and precision of measurement of these rare events should facilitate measurements to characterize the latent HIV reservoir and interventions to eradicate it.
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Cites
High-throughput droplet digital PCR system for absolute quantitation of DNA copy number
Hindson, B. J., Ness, K. D., Masquelier, D. A., Belgrader, P., Heredia, N. J., Makarewicz, A. J., Bright, I. J., Lucero, M. Y., Hiddessen, A. L., Legler, T. C., et al. (2011) · Analytical Chemistry · doi:10.1021/ac202028g · Cited by 2,792
Vogelstein, B., & Kinzler, K. W. (1999) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.96.16.9236 · Cited by 1,770
Whale, A. S., Huggett, J. F., Cowen, S., Speirs, V., Shaw, J., Ellison, S., Foy, C. A., & Scott, D. J. (2012) · Nucleic Acids Research · doi:10.1093/nar/gks203 · Cited by 453 · cited 1×
“Digital PCR (dPCR) [2], in which the amplification reaction is divided into thousands of microscopic reaction volumes, is a rapidly growing alternative that is potentially more accurate [3,4,5] and more precise [6,7].”
BackgroundKreutz, J. E., Munson, T., Huynh, T., Shen, F., Du, W., & Ismagilov, R. F. (2011) · Analytical Chemistry · doi:10.1021/ac201658s · Cited by 148
Cited by
Defective proviruses rapidly accumulate during acute HIV-1 infection
Bruner, K. M., Murray, A. J., Pollack, R. A., Soliman, M. G., Laskey, S. B., Capoferri, A. A., Lai, J., Strain, M. C., Lada, S. M., Hoh, R., et al. (2016) · Nature Medicine · doi:10.1038/nm.4156 · Cited by 737
Self-citationRapid pathogen detection by metagenomic next-generation sequencing of infected body fluids
Gu, W., Deng, X., Lee, M., Sucu, Y. D., Arevalo, S., Stryke, D., Federman, S., Gopez, A., Reyes, K., Zorn, K., et al. (2021) · Nature Medicine · doi:10.1038/s41591-020-1105-z · Cited by 689
Quan, P.-L., Sauzade, M., & Brouzes, E. (2018) · Sensors · doi:10.3390/s18041271 · Cited by 664
Comparative Analysis of Measures of Viral Reservoirs in HIV-1 Eradication Studies
Eriksson, S., Graf, E. H., Dahl, V., Strain, M. C., Yukl, S. A., Lysenko, E. S., Bosch, R. J., Lai, J., Chioma, S., Emad, F., et al. (2013) · PLoS Pathogens · doi:10.1371/journal.ppat.1003174 · Cited by 567
Self-citationConsiderations for Digital PCR as an Accurate Molecular Diagnostic Tool
Huggett, J. F., Cowen, S., & Foy, C. A. (2015) · Clinical Chemistry · doi:10.1373/clinchem.2014.221366 · Cited by 444
Metadata
- Authors
- Strain, M. C., Lada, S. M., Luong, T., Rought, S. E., Gianella, S., Terry, V. H., Spina, C. A., Woelk, C. H., & Richman, D. D.
- Venue
- PLoS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2013-04-03
- Volume
- 8
- Issue
- 4
- Pages
- e55943
- ISSN
- 1932-6203 (ISSN-L)
- OpenAlex
- W2128446740
- Cited by
- 504
- References
- 4
- FWCI
- 21.83
- Type
- journal article
- Language
- en
- Open access
- gold
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- Strain, M. C., Lada, S. M., Luong, T., Rought, S. E., Gianella, S., Terry, V. H., Spina, C. A., Woelk, C. H., & Richman, D. D. (2013). Highly Precise Measurement of HIV DNA by Droplet Digital PCR. PLoS ONE. https://doi.org/10.1371/journal.pone.0055943
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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