HIV-1 western blot assay: What determines an indeterminate status?

Balakrishnan, Pachamuthu; Solomon, SunilS; Murugavel, K; Kumarasamy, N.; Vidya, S.; Martin, SP; Thyagarajan, S P; Mayer, KennethH; Solomon, Suniti; Syed, IqbalH

doi:10.4103/0019-5359.17051

Cited by 14 publications

(2 citation statements)

References 17 publications

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“…42 Along with CD4+ lymphocyte count, these techniques include total lymphocyte count, HIV plasma viral load (PVL) and other surrogate markers. 46,47 Initial work was targeted at validating and optimising commercial manual assays such as the Coulter Cytosphere assay for resource poor settings. 48,49 However, recognising of the potential of microfluidics and bioMEMs 50 in biomedical diagnostics, much effort has been made to adapt these strategies as a viable option for low-cost HIV diagnosis in resource-poor settings.…”

Section: Future Pipeline and Market For Poc Cd4-based Hiv Diagnosticsmentioning

confidence: 99%

CD4 counting technologies for HIV therapy monitoring in resource-poor settings – state-of-the-art and emerging microtechnologies

2013

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Modern advancements in pharmaceuticals have provided individuals who have been infected with the human immunodeficiency virus (HIV) with the possibility of significantly extending their survival rates. When administered sufficiently soon after infection, antiretroviral therapy (ART) allows medical practitioners to control onset of the symptoms of the associated acquired immune deficiency syndrome (AIDS). Active monitoring of the immune system in both HIV patients and individuals who are regarded as "at-risk" is critical in the decision making process for when to start a patient on ART. A reliable and common method for such monitoring is to observe any decline in the number of CD4 expressing T-helper cells in the blood of a patient. However, the technology, expertise, infrastructure and costs to carry out such a diagnostic cannot be handled by medical services in resource-poor regions where HIV is endemic. Addressing this shortfall, commercialized point-of-care (POC) CD4 cell count systems are now available in such regions. A number of newer devices will also soon be on the market, some the result of recent maturing of charity-funded initiatives. Many of the current and imminent devices are enabled by microfluidic solutions, and this review will critically survey and analyze these POC technologies for CD4 counting, both on-market and near-to-market deployment. Additionally, promising technologies under development that may usher in a new generation of devices will be presented.

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Section: Future Pipeline and Market For Poc Cd4-based Hiv Diagnosticsmentioning

confidence: 99%

CD4 counting technologies for HIV therapy monitoring in resource-poor settings – state-of-the-art and emerging microtechnologies

2013

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“…NACO recommends the use of Western Blot assay/Line Immuno assay, for which the sample has to be referred to the National reference laboratory (NRL). Indeterminate results hinder clinical decisions, create more work, take up more resources in laboratories, and cause inappropriate anxiety to patients or even undesirable consequences [6] [7]. Hence, the present study was conducted to determine the utility of the current diagnostic algorithm in correctly identifying HIV infections and to find out the proportion of indeterminate or discrepant results at our centre over a period of five years.…”

Section: Introductionmentioning

confidence: 99%

HIV Diagnosis in Resource Constrained Setting: How Good Is the Current Algorithm?

Ingole¹,

Sarkate²,

Paranjpe³

et al. 2014

WJA

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Background: The conventional HIV testing algorithm in most of the developed countries consists of two tests: an HIV enzyme immunoassay capable of identifying HIV-1 and HIV-2 antibodies and a confirmatory HIV-1 Western blot or immunofluorescence assay. However, the current algorithm for HIV diagnosis in India uses three sequential antibody assays. There has always been doubt regarding the benefits of this algorithm. Objective: To determine the utility of the current diagnostic algorithm and to find out the proportion of indeterminate or discrepant results. Methods: Retrospective analysis of HIV antibody testing data was carried out over a period of five years after institutional ethics committee approval. The specimens positive with the screening test and negative with both the supplemental tests were labeled as discrepant. Specimens positive with any of the two tests (screening and one of the supplemental tests) and negative with the remaining supplemental test were labeled as indeterminate. These indeterminate specimens were confirmed by immunoblotting. Results: A total of 141,296 samples were tested. Of these, 71 (0.05%) samples were indeterminate and 292 (0.21%) were discrepant. Western blot was done on 60 indeterminate samples of which 10 (16.67%) were positive for HIV 1 antibodies, 14 (23.33%) were negative for HIV antibodies and 36 (60%) had indeterminate result. Conclusion: In view of the low numbers of indeterminate and discrepant results, the current algorithm appears to be appropriate in our resource constrained setting. However, the algorithm for HIV testing should also include DNA PCR testing facility to resolve western blot indeterminate results.

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Multistrip Western blotting to increase quantitative data output

et al. 2007

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The qualitative and quantitative measurement of protein abundance and protein modification states are essential in understanding their role in diverse cellular processes. Traditional Western blotting technique, though sensitive, is prone to produce substantial errors and is not readily adapted to highthroughput technologies. We propose a modified immunoblotting procedure, which is based on simultaneous transfer of proteins from multiple gel-strips onto the same membrane, and is compatible with any conventional gel electrophoresis system. As a result, the data output per single blotting cycle can readily be increased up to ten-fold. In contrast to the traditional "one protein detection per electrophoresis cycle", this procedure allows simultaneous monitoring of up to nine different proteins. In addition to maintaining the ability to detect picogram quantities of protein, the modified system substantially improves data accuracy by reducing signal errors by two-fold. Multi-strip Western blotting procedure allows making statistically reliable side-by-side comparisons of different or repeated sets of data. Compared to the traditional methods, this approach provides a more economical, reproducible and effective procedure, facilitating the generation of large amounts of high-quality quantifiable data.

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HIV-1 western blot assay: What determines an indeterminate status?

Cited by 14 publications

References 17 publications

CD4 counting technologies for HIV therapy monitoring in resource-poor settings – state-of-the-art and emerging microtechnologies

CD4 counting technologies for HIV therapy monitoring in resource-poor settings – state-of-the-art and emerging microtechnologies

HIV Diagnosis in Resource Constrained Setting: How Good Is the Current Algorithm?

Multistrip Western blotting to increase quantitative data output

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