Clinical evaluation of a low cost, in-house developed real-time RT-PCR human immunodeficiency virus type 1 (HIV-1) quantitation assay for HIV-1 infected patients.
OBJECTIVES: HIV-1 viral quantitation is essential for treatment monitoring. An in-house assay would decrease financial barriers to access. MATERIALS AND METHODS: A real-time competitive RT-PCR in house assay (Sing-IH) was developed in Singapore. Using HXB2 as reference, the assay's primers and...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3945479?pdf=render |
| _version_ | 1819226704685563904 |
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| author | Palvinder Kaur Wei Xin Khong Sue Yuen Wee Eng Lee Tan Juergen Pipper Evelyn Koay Kah Ying Ng Joe Kwan Yap Kuan Kiat Chew Mei Ting Tan Yee Sin Leo Masafumi Inoue Oon Tek Ng |
| author_facet | Palvinder Kaur Wei Xin Khong Sue Yuen Wee Eng Lee Tan Juergen Pipper Evelyn Koay Kah Ying Ng Joe Kwan Yap Kuan Kiat Chew Mei Ting Tan Yee Sin Leo Masafumi Inoue Oon Tek Ng |
| author_sort | Palvinder Kaur |
| collection | DOAJ |
| description | OBJECTIVES: HIV-1 viral quantitation is essential for treatment monitoring. An in-house assay would decrease financial barriers to access. MATERIALS AND METHODS: A real-time competitive RT-PCR in house assay (Sing-IH) was developed in Singapore. Using HXB2 as reference, the assay's primers and probes were designed to generate a 183-bp product that overlaps a portion of the LTR region and gag region. A competitive internal control (IC) was included in each assay to monitor false negative results due to inhibition or human error. Clinical evaluation was performed on 249 HIV-1 positive patient samples in comparison with the commercially available Generic HIV Viral Load assay. Correlation and agreement of results were assessed for plasma HIV-1 quantification with both assays. RESULTS: The assay has a lower limit of detection equivalent to 126 copies/mL of HIV-1 RNA and a linear range of detection from 100-1000000 copies/mL. Comparative analysis with reference to the Generic assay demonstrated good agreement between both assays with a mean difference of 0.22 log10 copies/mL and 98.8% of values within 1 log10 copies/mL range. Furthermore, the Sing-IH assay can quantify HIV-1 group M subtypes A-H and group N isolates adequately, making it highly suitable for our region, where subtype B and CRF01_AE predominate. CONCLUSIONS: With a significantly lower running cost compared to commercially available assays, the broadly sensitive Sing-IH assay could help to overcome the cost barriers and serve as a useful addition to the currently limited HIV viral load assay options for resource-limited settings. |
| first_indexed | 2024-12-23T10:29:43Z |
| format | Article |
| id | doaj.art-f0ff80979bbf4ba8b042ddf9031e995b |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-23T10:29:43Z |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-f0ff80979bbf4ba8b042ddf9031e995b2022-12-21T17:50:28ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0193e8982610.1371/journal.pone.0089826Clinical evaluation of a low cost, in-house developed real-time RT-PCR human immunodeficiency virus type 1 (HIV-1) quantitation assay for HIV-1 infected patients.Palvinder KaurWei Xin KhongSue Yuen WeeEng Lee TanJuergen PipperEvelyn KoayKah Ying NgJoe Kwan YapKuan Kiat ChewMei Ting TanYee Sin LeoMasafumi InoueOon Tek NgOBJECTIVES: HIV-1 viral quantitation is essential for treatment monitoring. An in-house assay would decrease financial barriers to access. MATERIALS AND METHODS: A real-time competitive RT-PCR in house assay (Sing-IH) was developed in Singapore. Using HXB2 as reference, the assay's primers and probes were designed to generate a 183-bp product that overlaps a portion of the LTR region and gag region. A competitive internal control (IC) was included in each assay to monitor false negative results due to inhibition or human error. Clinical evaluation was performed on 249 HIV-1 positive patient samples in comparison with the commercially available Generic HIV Viral Load assay. Correlation and agreement of results were assessed for plasma HIV-1 quantification with both assays. RESULTS: The assay has a lower limit of detection equivalent to 126 copies/mL of HIV-1 RNA and a linear range of detection from 100-1000000 copies/mL. Comparative analysis with reference to the Generic assay demonstrated good agreement between both assays with a mean difference of 0.22 log10 copies/mL and 98.8% of values within 1 log10 copies/mL range. Furthermore, the Sing-IH assay can quantify HIV-1 group M subtypes A-H and group N isolates adequately, making it highly suitable for our region, where subtype B and CRF01_AE predominate. CONCLUSIONS: With a significantly lower running cost compared to commercially available assays, the broadly sensitive Sing-IH assay could help to overcome the cost barriers and serve as a useful addition to the currently limited HIV viral load assay options for resource-limited settings.http://europepmc.org/articles/PMC3945479?pdf=render |
| spellingShingle | Palvinder Kaur Wei Xin Khong Sue Yuen Wee Eng Lee Tan Juergen Pipper Evelyn Koay Kah Ying Ng Joe Kwan Yap Kuan Kiat Chew Mei Ting Tan Yee Sin Leo Masafumi Inoue Oon Tek Ng Clinical evaluation of a low cost, in-house developed real-time RT-PCR human immunodeficiency virus type 1 (HIV-1) quantitation assay for HIV-1 infected patients. PLoS ONE |
| title | Clinical evaluation of a low cost, in-house developed real-time RT-PCR human immunodeficiency virus type 1 (HIV-1) quantitation assay for HIV-1 infected patients. |
| title_full | Clinical evaluation of a low cost, in-house developed real-time RT-PCR human immunodeficiency virus type 1 (HIV-1) quantitation assay for HIV-1 infected patients. |
| title_fullStr | Clinical evaluation of a low cost, in-house developed real-time RT-PCR human immunodeficiency virus type 1 (HIV-1) quantitation assay for HIV-1 infected patients. |
| title_full_unstemmed | Clinical evaluation of a low cost, in-house developed real-time RT-PCR human immunodeficiency virus type 1 (HIV-1) quantitation assay for HIV-1 infected patients. |
| title_short | Clinical evaluation of a low cost, in-house developed real-time RT-PCR human immunodeficiency virus type 1 (HIV-1) quantitation assay for HIV-1 infected patients. |
| title_sort | clinical evaluation of a low cost in house developed real time rt pcr human immunodeficiency virus type 1 hiv 1 quantitation assay for hiv 1 infected patients |
| url | http://europepmc.org/articles/PMC3945479?pdf=render |
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