Comparison of an in-house 'home-brew' and commercial ViroSeq integrase genotyping assays on HIV-1 subtype C samples.
<h4>Background</h4>Roll-out of Integrase Strand Transfer Inhibitors (INSTIs) such as dolutegravir for HIV combination antiretroviral therapy (cART) in sub-Saharan Africa necessitates the development of affordable HIV drug resistance (HIVDR) assays targeting the Integrase gene. We optimis...
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doaj-563981b73ee344e398ad6377ba9529912021-03-04T12:43:26ZengPublic Library of Science (PLoS)PLoS ONE1932-62032019-01-011411e022429210.1371/journal.pone.0224292Comparison of an in-house 'home-brew' and commercial ViroSeq integrase genotyping assays on HIV-1 subtype C samples.Kaelo K SeatlaWonderful T ChogaMompati MogweleThabo DiphokoDorcas MaruapulaLucy MupfumiRosemary M MusondaChristopher F RowleyAva AvalosIshmael KasvosveSikhulile MoyoSimani Gaseitsiwe<h4>Background</h4>Roll-out of Integrase Strand Transfer Inhibitors (INSTIs) such as dolutegravir for HIV combination antiretroviral therapy (cART) in sub-Saharan Africa necessitates the development of affordable HIV drug resistance (HIVDR) assays targeting the Integrase gene. We optimised and evaluated an in-house integrase HIV-1 drug resistance assay (IH-Int) and compared it to a commercially available assay, ViroSeq™ Integrase Genotyping kit (VS-Int) amongst HIV-1 clade C infected individuals.<h4>Methods</h4>We used 54 plasma samples from treatment naïve participants and one plasma sample from a patient failing INSTI based cART. Specimens were genotyped using both the VS-Int and IH-Int assays. Stanford HIV drug resistance database were used for integrase resistance interpretation. We compared the major and minor resistance mutations, pairwise nucleotide and amino-acid identity, costs and assay time.<h4>Results</h4>Among 55 specimens tested with IH-Int, 53 (96.4%) successfully amplified compared to 45/55 (81.8%) for the VS-Int assay. The mean nucleotide and amino acid similarity from 33 paired sequences was 99.8% (SD ± 0.30) and 99.8% (SD ± 0.39) for the IH-Int and VS-Int assay respectively. The reagent cost/sample were 32 USD and 147 USD for IH-Int and VS-Int assay, respectively. All sequenced samples were confirmed as HIV-1 subtype C.<h4>Conclusions</h4>The IH-Int assay had a high amplification success rate and high concordance with the commercial assay. It is significantly cheaper compared to the commercial assay. Our assay has the needed specifications for routine monitoring of participants on Dolutegravir based regimens in Botswana.https://doi.org/10.1371/journal.pone.0224292 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Kaelo K Seatla Wonderful T Choga Mompati Mogwele Thabo Diphoko Dorcas Maruapula Lucy Mupfumi Rosemary M Musonda Christopher F Rowley Ava Avalos Ishmael Kasvosve Sikhulile Moyo Simani Gaseitsiwe |
spellingShingle |
Kaelo K Seatla Wonderful T Choga Mompati Mogwele Thabo Diphoko Dorcas Maruapula Lucy Mupfumi Rosemary M Musonda Christopher F Rowley Ava Avalos Ishmael Kasvosve Sikhulile Moyo Simani Gaseitsiwe Comparison of an in-house 'home-brew' and commercial ViroSeq integrase genotyping assays on HIV-1 subtype C samples. PLoS ONE |
author_facet |
Kaelo K Seatla Wonderful T Choga Mompati Mogwele Thabo Diphoko Dorcas Maruapula Lucy Mupfumi Rosemary M Musonda Christopher F Rowley Ava Avalos Ishmael Kasvosve Sikhulile Moyo Simani Gaseitsiwe |
author_sort |
Kaelo K Seatla |
title |
Comparison of an in-house 'home-brew' and commercial ViroSeq integrase genotyping assays on HIV-1 subtype C samples. |
title_short |
Comparison of an in-house 'home-brew' and commercial ViroSeq integrase genotyping assays on HIV-1 subtype C samples. |
title_full |
Comparison of an in-house 'home-brew' and commercial ViroSeq integrase genotyping assays on HIV-1 subtype C samples. |
title_fullStr |
Comparison of an in-house 'home-brew' and commercial ViroSeq integrase genotyping assays on HIV-1 subtype C samples. |
title_full_unstemmed |
Comparison of an in-house 'home-brew' and commercial ViroSeq integrase genotyping assays on HIV-1 subtype C samples. |
title_sort |
comparison of an in-house 'home-brew' and commercial viroseq integrase genotyping assays on hiv-1 subtype c samples. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2019-01-01 |
description |
<h4>Background</h4>Roll-out of Integrase Strand Transfer Inhibitors (INSTIs) such as dolutegravir for HIV combination antiretroviral therapy (cART) in sub-Saharan Africa necessitates the development of affordable HIV drug resistance (HIVDR) assays targeting the Integrase gene. We optimised and evaluated an in-house integrase HIV-1 drug resistance assay (IH-Int) and compared it to a commercially available assay, ViroSeq™ Integrase Genotyping kit (VS-Int) amongst HIV-1 clade C infected individuals.<h4>Methods</h4>We used 54 plasma samples from treatment naïve participants and one plasma sample from a patient failing INSTI based cART. Specimens were genotyped using both the VS-Int and IH-Int assays. Stanford HIV drug resistance database were used for integrase resistance interpretation. We compared the major and minor resistance mutations, pairwise nucleotide and amino-acid identity, costs and assay time.<h4>Results</h4>Among 55 specimens tested with IH-Int, 53 (96.4%) successfully amplified compared to 45/55 (81.8%) for the VS-Int assay. The mean nucleotide and amino acid similarity from 33 paired sequences was 99.8% (SD ± 0.30) and 99.8% (SD ± 0.39) for the IH-Int and VS-Int assay respectively. The reagent cost/sample were 32 USD and 147 USD for IH-Int and VS-Int assay, respectively. All sequenced samples were confirmed as HIV-1 subtype C.<h4>Conclusions</h4>The IH-Int assay had a high amplification success rate and high concordance with the commercial assay. It is significantly cheaper compared to the commercial assay. Our assay has the needed specifications for routine monitoring of participants on Dolutegravir based regimens in Botswana. |
url |
https://doi.org/10.1371/journal.pone.0224292 |
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