A Dried Blood Spot Update: Still an Important Bioanalytical Technique?

Spooner, Neil

doi:10.4155/bio.13.56

Cited by 30 publications

(14 citation statements)

References 42 publications

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“…However, as more research was undertaken to underpin the science behind DBSs, a number of uncertainties arose, such as the homogeneity of DBS samples, the optimum method of adding IS and performing dilutions, the stability of aged DBSs, and most notably the impact of HCT on assay bias (19) . The FDA currently recommend that, until instructed otherwise, DBS data to support drug development studies should be run in parallel with conventionally used wet plasma or whole blood sampling (123,29,23) . It is assumed that the FDA will only recommend DBS data to be used in isolation once they have received, monitored and approved enough drug development studies for them to have sufficient confidence that the technique produces reliable and accurate data (123,29,23) .…”

Section: Overall Conclusionmentioning

confidence: 99%

“…The FDA currently recommend that, until instructed otherwise, DBS data to support drug development studies should be run in parallel with conventionally used wet plasma or whole blood sampling (123,29,23) . It is assumed that the FDA will only recommend DBS data to be used in isolation once they have received, monitored and approved enough drug development studies for them to have sufficient confidence that the technique produces reliable and accurate data (123,29,23) . Unfortunately the requirement to generate parallel wet and dry data has a significant cost implication (59) , and consequently the interest in, and momentum behind using DBSs for drug development has dropped significantly as project managers in pharmaceutical development were not able to justify the extra costs (except in niche applications where other sampling alternatives were not possible).…”

Section: Overall Conclusionmentioning

confidence: 99%

“…Unfortunately the momentum behind introducing DBS analysis as a widely used technique into highly regulated applications (such as pharmaceutical drug development) has recently been slowed due to a number of barriers. These include uncertainty over regulatory acceptance, scientific uncertainty over assay bias caused by varying haematocrit (HCT), and also concerns relating to the increased complexity of extraction, and lower assay sensitivity associated with DBS analysis (123,70) . To try and counter some of these issues our group has recently carried out research on nullifying haematocrit (HCT) based assay bias, and maximising assay sensitivity via the development of direct elution techniques in dried blood spot (DBS) quantitative bioanalysis (81,124) .…”

Section: Introductionmentioning

confidence: 99%

“…However, recently the momentum behind introducing DBS analysis into highly regulated applications (such as pharmaceutical drug development) has hit a significant bottleneck due to a number of factors. The primary issues are uncertainty over regulatory acceptance, and scientific uncertainty over assay bias caused by varying haematocrit (HCT); while secondary issues include barriers to bioanalytical acceptance caused by the increased complexity of extraction, and lower assay sensitivity associated with DBS analysis (123,70,23) .…”

Section: Introductionmentioning

confidence: 99%

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Direct Analysis of Dried Blood Spot Samples

Abu-Rabie¹

2014

Dried Blood Spots

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Section: Overall Conclusionmentioning

confidence: 99%

Section: Overall Conclusionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Direct Analysis of Dried Blood Spot Samples

Abu-Rabie¹

2014

Dried Blood Spots

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“…Dried blood samples spotted on filter papers, introduced by Robert Guthrie in 1963, have been widely used in neonatal screening programs for the early identification of congenital disorders and presymptomatic management of affected neonates [1][2][3]. The DBS were first used in the early 1970s in the neonatal screening for phenylketonuria (PKU) which provided a simple, inexpensive and unique method for heel-stick blood collection from neonates onto a special cotton fiber filter-paper which is still known as a "PKU card" or "Guthrie card" [4,5].…”

Section: Introductionmentioning

confidence: 99%

Superparamagnetic-bead Based Method: An Effective DNA Extraction from Dried Blood Spots (DBS) for Diagnostic PCR

Sirdah¹

2014

JCDR

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IntrOductIOnDried blood samples spotted on filter papers, introduced by Robert Guthrie in 1963, have been widely used in neonatal screening programs for the early identification of congenital disorders and presymptomatic management of affected neonates [1][2][3]. The DBS were first used in the early 1970s in the neonatal screening for phenylketonuria (PKU) which provided a simple, inexpensive and unique method for heel-stick blood collection from neonates onto a special cotton fiber filter-paper which is still known as a "PKU card" or "Guthrie card" [4,5]. Since then, DBS have been collected routinely from babies not only for PKU screening but also for other biochemical screenings associated with congenital, inherited, and infectious disorders [6][7][8]. The success of these DBS as an easy and cost-effective method for collecting, archiving, and transporting blood specimens has inspired researchers to explore better DNA extraction and PCR-based molecular testing methods for these samples [9][10][11][12]. The DBS sampling is an effective alternative for lymphocyte pellets because DBS sampling requires only a few droplets of whole blood, not necessarily venous blood, and can be directly spotted on Guthrie cards or filter papers [13][14][15].The commonly used methods and techniques for extracting genomic DNA from DBS are varied in terms of extraction principle, reagents used, and quantity and quality of the extracted DNA. Manual protocols include the extraction of DNA with Tris-EDTA buffer, methanol, and Chelex-100 [16][17][18][19]. In addition, commercially available kits for DNA extraction from DBS are available [11,20], but their cost could limit their use for large scale-screening programs.Paramagnetic-bead based extraction method of DNA from body fluids such as semen, blood, vaginal, and buccal cells has been established and resulted in a cost-efficient and high-throughput DNA extraction method that can be used for PCR-based molecular Pathology Section testing [21][22][23][24]. The present work explores and evaluates the effectiveness of a superparamagnetic-bead based method (LGC Genomic, Germany) in extracting DNA from DBS. MAterIAls And MethOdsWhole blood samples collected from 16 apparently healthy subjects and two ß-thalassemia patients were used for spotting onto filter papers, direct DNA extraction and quantitative and qualitative evaluations. spotting onto filter papersFrom each sample four 100 µl applications of whole blood were spotted onto Ahlstrom 226 grad newborn screening filter papers (ID Biological Systems, Greenville, SC, USA), and were dried at ambient temperature (28-30 0 C) for one week. dnA extractionDNA extraction from whole blood and DBS were performed using the superparamagnetic-bead based DNA extraction kit manufactured by LGC Genomic, formerly AGOWA, Germany. The extraction was carried out according to the manufacturer's instruction for whole blood and with some modifications for extraction from DBS. The standard superparamagnetic-bead based DNA extraction protocol includes four princi...

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Automated high throughput analysis of antiretroviral drugs in dried blood spots

et al. 2017

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For therapeutic drug monitoring in remote settings, dried blood spots (DBS) are particularly advantageous, as blood sample collection and handling is uncomplicated. The aim of this study was to develop and validate an automated extraction method for the analysis of nevirapine, efavirenz and lopinavir in DBS samples. Automated extraction was performed with methanol : water (70 : 30 v/v), using a DBS-MS 500 autosampler coupled to a liquid chromatography tandem mass spectrometry system. The autosampler used digital images of each DBS to position the extraction head, sprayed 10 μl of internal standard onto each DBS and extracted a 4-mm disc (Ø) from the centre of each spot by unilateral flow using 25-μl extraction solvent. The analytes were baseline separated on a pentafluorophenyl column and analysed by using electrospray ionization with multiple reaction monitoring in positive polarity mode for nevirapine and lopinavir and in negative mode for efavirenz. The method was linear between 10 and 10 000 ng/ml for all analytes. Automated sample extraction resulted in consistent recoveries (nevirapine: 70 ± 6%, efavirenz: 63 ± 11% and lopinavir: 60 ± 10%) and matrix effects between different donors and concentration levels. Intra-day and inter-day accuracy and precision deviations were ≤15%. Manual and automated extractions of DBS samples collected within the framework of an adherence assessment study in rural Tanzania showed good agreements with deviations of less than 10%. Our study highlights that therapeutic drug monitoring samples obtained in the resource-constrained setting of rural Africa can be reliably determined by automated extraction of DBS. Overall, automatization improved method sensitivity and facilitates analysis of large sample numbers. Copyright © 2017 John Wiley & Sons, Ltd.

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A Dried Blood Spot Update: Still an Important Bioanalytical Technique?

Cited by 30 publications

References 42 publications

Direct Analysis of Dried Blood Spot Samples

Direct Analysis of Dried Blood Spot Samples

Superparamagnetic-bead Based Method: An Effective DNA Extraction from Dried Blood Spots (DBS) for Diagnostic PCR

Automated high throughput analysis of antiretroviral drugs in dried blood spots

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