The development, evaluation, performance, and validation of micro-PCR and extractor for the quantification of HIV-1 &-2 RNA
Abstract HIV infection has been a global public health threat and overall reported ~ 40 million deaths. Acquired immunodeficiency syndrome (AIDS) is attributed to the retroviruses (HIV-1/2), disseminated through various body fluids. The temporal progression of AIDS is in context to the rate of HIV-1...
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| Format: | Article |
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Nature Portfolio
2024-04-01
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| Series: | Scientific Reports |
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| Online Access: | https://doi.org/10.1038/s41598-024-56164-5 |
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| author | Shyam Prakash Ram Aasarey Priyatma Meenakshi Sharma Shahid Khan Medha |
| author_facet | Shyam Prakash Ram Aasarey Priyatma Meenakshi Sharma Shahid Khan Medha |
| author_sort | Shyam Prakash |
| collection | DOAJ |
| description | Abstract HIV infection has been a global public health threat and overall reported ~ 40 million deaths. Acquired immunodeficiency syndrome (AIDS) is attributed to the retroviruses (HIV-1/2), disseminated through various body fluids. The temporal progression of AIDS is in context to the rate of HIV-1 infection, which is twice as protracted in HIV-2 transmission. Q-PCR is the only available method that requires a well-developed lab infrastructure and trained personnel. Micro-PCR, a portable Q-PCR device, was developed by Bigtec Labs, Bangalore, India. It is simple, accurate, fast, and operationalised in remote places where diagnostic services are inaccessible in developing countries. This novel micro-PCR determines HIV-1 and HIV-2 viral load using a TruePrep™ extractor device for RNA isolation. Five ml blood samples were collected at the blood collection centre at AIIMS, New Delhi, India. Samples were screened for serology, and a comparison of HIV-1/2 RNA was done between qPCR and micro-PCR in the samples. The micro-PCR assay of HIV-RNA has compared well with those from real-time PCR (r = 0.99, i < 0.002). Micro-PCR has good inter and intra-assay reproducibility over a wide dynamic range (1.0 × 102–1.0 × 108 IU/ml). The linear dynamic range was 102–108 IU/ml. The clinical and analytical specificity of the assay was comparable, i.e., 100%. Intra-assay and inter-assay coefficients of variation ranged from 1.17% to 3.15% and from 0.02% to 0.46%, respectively. Moreover, due to the robust, simple, and empirical method, the Probit analysis has also been done for qPCR LODs to avoid uncertainties in target recoveries. The micro-PCR is reliable, accurate, and reproducible for early detection of HIV-1 and HIV-2 viral loads simultaneously. Thus, it can easily be used in the field and in remote places where quantification of both HIV-1/2 is not reachable. |
| first_indexed | 2024-04-24T07:16:50Z |
| format | Article |
| id | doaj.art-e34fdaf75eb54f119042b997d4fe12af |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-24T07:16:50Z |
| publishDate | 2024-04-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-e34fdaf75eb54f119042b997d4fe12af2024-04-21T11:17:13ZengNature PortfolioScientific Reports2045-23222024-04-0114111010.1038/s41598-024-56164-5The development, evaluation, performance, and validation of micro-PCR and extractor for the quantification of HIV-1 &-2 RNAShyam Prakash0Ram Aasarey1Priyatma2Meenakshi Sharma3Shahid Khan4Medha5Department of Laboratory Medicine, All India Institute of Medical SciencesDepartment of Laboratory Medicine, All India Institute of Medical SciencesDepartment of Laboratory Medicine, All India Institute of Medical SciencesDepartment of Laboratory Medicine, All India Institute of Medical SciencesDepartment of Laboratory Medicine, All India Institute of Medical SciencesDepartment of Laboratory Medicine, All India Institute of Medical SciencesAbstract HIV infection has been a global public health threat and overall reported ~ 40 million deaths. Acquired immunodeficiency syndrome (AIDS) is attributed to the retroviruses (HIV-1/2), disseminated through various body fluids. The temporal progression of AIDS is in context to the rate of HIV-1 infection, which is twice as protracted in HIV-2 transmission. Q-PCR is the only available method that requires a well-developed lab infrastructure and trained personnel. Micro-PCR, a portable Q-PCR device, was developed by Bigtec Labs, Bangalore, India. It is simple, accurate, fast, and operationalised in remote places where diagnostic services are inaccessible in developing countries. This novel micro-PCR determines HIV-1 and HIV-2 viral load using a TruePrep™ extractor device for RNA isolation. Five ml blood samples were collected at the blood collection centre at AIIMS, New Delhi, India. Samples were screened for serology, and a comparison of HIV-1/2 RNA was done between qPCR and micro-PCR in the samples. The micro-PCR assay of HIV-RNA has compared well with those from real-time PCR (r = 0.99, i < 0.002). Micro-PCR has good inter and intra-assay reproducibility over a wide dynamic range (1.0 × 102–1.0 × 108 IU/ml). The linear dynamic range was 102–108 IU/ml. The clinical and analytical specificity of the assay was comparable, i.e., 100%. Intra-assay and inter-assay coefficients of variation ranged from 1.17% to 3.15% and from 0.02% to 0.46%, respectively. Moreover, due to the robust, simple, and empirical method, the Probit analysis has also been done for qPCR LODs to avoid uncertainties in target recoveries. The micro-PCR is reliable, accurate, and reproducible for early detection of HIV-1 and HIV-2 viral loads simultaneously. Thus, it can easily be used in the field and in remote places where quantification of both HIV-1/2 is not reachable.https://doi.org/10.1038/s41598-024-56164-5Micro-PCRHIV-1HIV-2Truenat™TruePrep™ |
| spellingShingle | Shyam Prakash Ram Aasarey Priyatma Meenakshi Sharma Shahid Khan Medha The development, evaluation, performance, and validation of micro-PCR and extractor for the quantification of HIV-1 &-2 RNA Scientific Reports Micro-PCR HIV-1 HIV-2 Truenat™ TruePrep™ |
| title | The development, evaluation, performance, and validation of micro-PCR and extractor for the quantification of HIV-1 &-2 RNA |
| title_full | The development, evaluation, performance, and validation of micro-PCR and extractor for the quantification of HIV-1 &-2 RNA |
| title_fullStr | The development, evaluation, performance, and validation of micro-PCR and extractor for the quantification of HIV-1 &-2 RNA |
| title_full_unstemmed | The development, evaluation, performance, and validation of micro-PCR and extractor for the quantification of HIV-1 &-2 RNA |
| title_short | The development, evaluation, performance, and validation of micro-PCR and extractor for the quantification of HIV-1 &-2 RNA |
| title_sort | development evaluation performance and validation of micro pcr and extractor for the quantification of hiv 1 2 rna |
| topic | Micro-PCR HIV-1 HIV-2 Truenat™ TruePrep™ |
| url | https://doi.org/10.1038/s41598-024-56164-5 |
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