Performance evaluation of a laboratory developed PCR test for quantitation of HIV-2 viral RNA.
Management of Human Immunodeficiency Virus Type 2 (HIV-2) infections present unique challenges due to low viral titers, slow disease progression, and poor response to standard antiviral therapies. The need for a nucleic acid assay to detect and quantify HIV-2 virus has led to the development of a nu...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2020-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0229424 |
| _version_ | 1818588602358038528 |
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| author | Linda L Jagodzinski Mark M Manak Holly R Hack Ying Liu Sheila A Peel |
| author_facet | Linda L Jagodzinski Mark M Manak Holly R Hack Ying Liu Sheila A Peel |
| author_sort | Linda L Jagodzinski |
| collection | DOAJ |
| description | Management of Human Immunodeficiency Virus Type 2 (HIV-2) infections present unique challenges due to low viral titers, slow disease progression, and poor response to standard antiviral therapies. The need for a nucleic acid assay to detect and quantify HIV-2 virus has led to the development of a number of molecular-based assays for detection and/or quantification of HIV-2 viral RNA in plasma in order to provide laboratory evidence of HIV-2 infection and viral loads for use in treatment decisions. As HIV-2 is less pathogenic and transmissible than HIV-1 and has resistance to several of the antiretroviral drugs, delay of treatment is common. Cross sero-reactivity between HIV-1 and HIV-2 makes it difficult to distinguish between the two viruses based upon serological tests. As such we developed a quantitative reverse transcription PCR (qRT-PCR) assay targeting the 5' long terminal repeat of HIV-2 for detection and quantification of HIV-2 viral RNA in plasma to identify HIV-2 infection and for use in viral load monitoring. Serial dilutions of cultured HIV-2 virus demonstrated a wide dynamic range (10 to 100,000 copies/ml) with excellent reproducibility (standard deviation from 0.12-0.19), linearity (R2 = 0.9994), and a lower limit of detection at 79 copies/ml (NIH-Z). The assay is highly specific for HIV-2 Groups A and B and exhibits no cross reactivity to HIV-1, HBV or HCV. Precision of the assay was demonstrated for the High (Mean = 6.41; SD = 0.12) and Medium (Mean = 4.46; SD = 0.13) HIV-2 positive controls. Replicate testing of clinical specimens showed good reproducibility above 1,000 copies/ml, with higher variability under 1,000 copies/ml. Analysis of 220 plasma samples from HIV-2 infected West African individuals demonstrated significantly lower viral loads than those observed in HIV-1 infections, consistent with results of previous studies. Slightly more than seven percent of clinical samples (7.3%) demonstrated viral loads above 100,000 copies/ml, while 37.3% of samples were undetectable. The high sensitivity, specificity, precision, and linearity of the WRAIR qRT-PCR assay makes it well suited for detection and monitoring of HIV-2 RNA levels in plasma of infected individuals. |
| first_indexed | 2024-12-16T09:27:22Z |
| format | Article |
| id | doaj.art-0b55bfc0558a41dea40b3892d0cb126f |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-16T09:27:22Z |
| publishDate | 2020-01-01 |
| publisher | Public Library of Science (PLoS) |
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| spelling | doaj.art-0b55bfc0558a41dea40b3892d0cb126f2022-12-21T22:36:36ZengPublic Library of Science (PLoS)PLoS ONE1932-62032020-01-01152e022942410.1371/journal.pone.0229424Performance evaluation of a laboratory developed PCR test for quantitation of HIV-2 viral RNA.Linda L JagodzinskiMark M ManakHolly R HackYing LiuSheila A PeelManagement of Human Immunodeficiency Virus Type 2 (HIV-2) infections present unique challenges due to low viral titers, slow disease progression, and poor response to standard antiviral therapies. The need for a nucleic acid assay to detect and quantify HIV-2 virus has led to the development of a number of molecular-based assays for detection and/or quantification of HIV-2 viral RNA in plasma in order to provide laboratory evidence of HIV-2 infection and viral loads for use in treatment decisions. As HIV-2 is less pathogenic and transmissible than HIV-1 and has resistance to several of the antiretroviral drugs, delay of treatment is common. Cross sero-reactivity between HIV-1 and HIV-2 makes it difficult to distinguish between the two viruses based upon serological tests. As such we developed a quantitative reverse transcription PCR (qRT-PCR) assay targeting the 5' long terminal repeat of HIV-2 for detection and quantification of HIV-2 viral RNA in plasma to identify HIV-2 infection and for use in viral load monitoring. Serial dilutions of cultured HIV-2 virus demonstrated a wide dynamic range (10 to 100,000 copies/ml) with excellent reproducibility (standard deviation from 0.12-0.19), linearity (R2 = 0.9994), and a lower limit of detection at 79 copies/ml (NIH-Z). The assay is highly specific for HIV-2 Groups A and B and exhibits no cross reactivity to HIV-1, HBV or HCV. Precision of the assay was demonstrated for the High (Mean = 6.41; SD = 0.12) and Medium (Mean = 4.46; SD = 0.13) HIV-2 positive controls. Replicate testing of clinical specimens showed good reproducibility above 1,000 copies/ml, with higher variability under 1,000 copies/ml. Analysis of 220 plasma samples from HIV-2 infected West African individuals demonstrated significantly lower viral loads than those observed in HIV-1 infections, consistent with results of previous studies. Slightly more than seven percent of clinical samples (7.3%) demonstrated viral loads above 100,000 copies/ml, while 37.3% of samples were undetectable. The high sensitivity, specificity, precision, and linearity of the WRAIR qRT-PCR assay makes it well suited for detection and monitoring of HIV-2 RNA levels in plasma of infected individuals.https://doi.org/10.1371/journal.pone.0229424 |
| spellingShingle | Linda L Jagodzinski Mark M Manak Holly R Hack Ying Liu Sheila A Peel Performance evaluation of a laboratory developed PCR test for quantitation of HIV-2 viral RNA. PLoS ONE |
| title | Performance evaluation of a laboratory developed PCR test for quantitation of HIV-2 viral RNA. |
| title_full | Performance evaluation of a laboratory developed PCR test for quantitation of HIV-2 viral RNA. |
| title_fullStr | Performance evaluation of a laboratory developed PCR test for quantitation of HIV-2 viral RNA. |
| title_full_unstemmed | Performance evaluation of a laboratory developed PCR test for quantitation of HIV-2 viral RNA. |
| title_short | Performance evaluation of a laboratory developed PCR test for quantitation of HIV-2 viral RNA. |
| title_sort | performance evaluation of a laboratory developed pcr test for quantitation of hiv 2 viral rna |
| url | https://doi.org/10.1371/journal.pone.0229424 |
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