Iohexol plasma clearance simplified by Dried Blood Spot (DBS) sampling to measure renal function in conscious mice

AbstractThere is no simple method to measure glomerular filtration rate (GFR) in mice, which limits the use of mice in models of renal diseases. We aimed at simplifying the plasma clearance of iohexol in mice, using dried blood spot (DBS) sampling in order to reduce the amount of blood taken for analysis. GFR was measured simultaneously by a reference method in total blood—as described before—and tested method using DBS in fifteen male and six female C57BL/6J mice. Total blood extraction was 50 μL for the reference methods and 25μL for the tested methods, distributed in 5 samples. The agreement of GFR values between both methods was analyzed with the concordance correlation coefficient (CCC), total deviation index (TDI) and coverage probability (CP). The agreement between both methods was excellent, showing a TDI = 8.1%, which indicates that 90% of the GFR values obtained with DBS showed an error ranging from − 8 to + 8% of the reference method; a CCC of 0.996 (CI: 0.992), reflecting high precision and accuracy and a CP of 94 (CI: 83), indicating that 6% of the GFR values obtained with DBS had an error greater than 10% of the method in blood. So, both methods are interchangeable. DBS represent a major simplification of GFR measurement in mice. Also, DBS improves animal welfare by reducing the total blood required and refining the procedure.

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A Simplified Iohexol-Based Method to Measure Renal Function in Sheep Models of Renal Disease

Biology ◽

10.3390/biology9090259 ◽

2020 ◽

Vol 9 (9) ◽

pp. 259

Author(s):

Sergio Luis-Lima ◽

Carolina Mas-Sanmartin ◽

Ana Elena Rodríguez-Rodríguez ◽

Esteban Porrini ◽

Alberto Ortiz ◽

...

Keyword(s):

Coverage Probability ◽

Filtration Rate ◽

Plasma Clearance ◽

Kidney Diseases ◽

Reference Method ◽

Renal Diseases ◽

Concordance Correlation ◽

Simple Method ◽

Correction Formula ◽

Animal Management

Sheep are highly adequate models for human renal diseases because of their many similarities in the histology and physiology of kidney and pathogenesis of kidney diseases. However, the lack of a simple method to measure glomerular filtration rate (GFR) limits its use as a model of renal diseases. Hence, we aimed to develop a simple method to measure GFR based on the plasma clearance of iohexol by assessing different pharmacokinetic models: (a) CL2: two-compartment (samples from 15 to 420 min; reference method); (b) CL1: one-compartment (samples from 60 to 420 min); (c) CLlf: CL1 adjusted by a correction formula and (d) SM: simplified CL2 (15 to 300 min). Specific statistics of agreement were used to test the models against CL2. The agreement between CL1 and CL2 was low, but both CL1f and SM showed excellent agreement with CL2, as indicated by a total deviation index of ~5–6%, a concordance correlation of 0.98–0.99% and a coverage probability of 99–100%, respectively. Hence, the SM approach is preferable due to a reduced number of samples and shorter duration of the procedure; two points that improve animal management and welfare.

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Iohexol plasma clearance simplified by dried blood spot testing

Nephrology Dialysis Transplantation ◽

10.1093/ndt/gfx323 ◽

2017 ◽

Cited By ~ 5

Author(s):

Sergio Luis-Lima ◽

Flavio Gaspari ◽

Natalia Negrín-Mena ◽

Fabiola Carrara ◽

Laura Díaz-Martín ◽

...

Keyword(s):

Plasma Clearance ◽

Dried Blood Spot ◽

Iohexol Plasma Clearance ◽

Blood Spot

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Assessment of dried blood spot samples as a simple method for detection of hepatitis B virus markers

Journal of Medical Virology ◽

10.1002/jmv.22138 ◽

2011 ◽

Vol 83 (9) ◽

pp. 1522-1529 ◽

Cited By ~ 45

Author(s):

Livia Melo Villar ◽

Jaqueline Correia de Oliveira ◽

Helena Medina Cruz ◽

Clara Fumiko Tachibana Yoshida ◽

Elisabeth Lampe ◽

...

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Dried Blood Spot ◽

Simple Method ◽

B Virus ◽

Hepatitis B Virus Markers ◽

Blood Spot

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Candidate reference method for determination of vitamin D from dried blood spot samples

Clinical Chemistry and Laboratory Medicine (CCLM) ◽

10.1515/cclm-2019-0397 ◽

2020 ◽

Vol 58 (5) ◽

pp. 817-827 ◽

Cited By ~ 2

Author(s):

Rosita Zakaria ◽

Katrina J. Allen ◽

Jennifer J. Koplin ◽

Peter Roche ◽

Ronda F. Greaves

Keyword(s):

Vitamin D ◽

Internal Standard ◽

Reference Method ◽

Population Level ◽

Population Based ◽

Dried Blood Spot ◽

Coefficient Of Determination ◽

Plasma Vitamin ◽

Blood Spot

AbstractBackgroundThe current millennium has seen an explosion in vitamin D testing with the overarching aim of requests to clinically stratify patients as replete or deficient in vitamin D. At a population level, dried blood spot (DBS) sampling offers a less invasive and more practical application for assessment of vitamin D status. We have therefore aimed to develop a sensitive and robust DBS vitamin D method that is traceable to serum for use in population-based studies.MethodsBlood spots, calibrators and controls were prepared by punching a 3.2 mm DBS from filter paper and placed into a 96-well micro-plate. The DBS disk was eluted with a combination of water-methanol and internal standard (ISTD) solution followed by supported-liquid extraction and derivatisation. The extract was analysed by liquid-chromatography tandem-mass spectrometry in positive electrospray-ionisation mode with 732.5 > 673.4 and 738.4 > 679.4 m/z ion-transitions for derivatised vitamin D and the ISTD, respectively. Vitamin D results were made traceable to the National Institute of Standards and Technology reference material through the inclusion of Chromsystems vitamin D calibrators.Results25-Hydroxy-vitamin D3 and its related ISTD were detected at a retention time of 7 min. The seven-point calibration-curve consistently demonstrated a coefficient of determination of 0.99 with an experimentally determined reportable range of 0.5–376 nmol/L. Method validation studies using DBS samples demonstrated 12.9% between-assay imprecision at 45 nmol/L, 84% average recovery and high correlation with plasma vitamin D (correlation coefficient = 0.86).ConclusionsWe have successfully developed an analytical method for vitamin D quantitation from DBSs which will be applied to our population-based vitamin D research study. This approach improves traceability of DBS results and potentially could be used broadly for other DBS measurands that require comparison to serum/plasma for their interpretation.

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Assessment of a dried blood spot C-reactive protein method to identify disease flares in rheumatoid arthritis patients

Scientific Reports ◽

10.1038/s41598-020-77826-0 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Leon G. D’Cruz ◽

Kevin G. McEleney ◽

Chris Cochrane ◽

Kyle B. C. Tan ◽

Priyank Shukla ◽

...

Keyword(s):

Rheumatoid Arthritis ◽

Joint Damage ◽

Reference Method ◽

Detection Sensitivity ◽

Dried Blood Spot ◽

Enzyme Linked Immunosorbent Assay ◽

C Reactive Protein ◽

Reactive Protein ◽

Deming Regression ◽

Blood Spot

AbstractRheumatoid arthritis (RA) is characterised by painful, stiff and swollen joints. RA features sporadic ‘flares’ or inflammatory episodes—mostly occurring outside clinics—where symptoms worsen and plasma C-reactive protein (CRP) becomes elevated. Poor control of inflammation results in higher rates of irreversible joint damage, increased disability, and poorer quality of life. Flares need to be accurately identified and managed. A method comparison study was designed to assess agreement between CRP values obtained by dried blood spot (DBS) versus conventional venepuncture sampling. The ability of a weekly DBS sampling and CRP test regime to detect flare outside the clinic was also assessed. Matched venepuncture and finger lancet DBS samples were collected from n = 100 RA patients with active disease at baseline and 6 weeks (NCT02809547). A subset of n = 30 RA patients submitted weekly DBS samples over the study period. Patient demographics, including self-reported flares were recorded. DBS sample CRP measurements were made by enzyme-linked immunosorbent assay, and venepuncture samples by a reference immunoturbometric assay. Data was compared between sample types by Bland–Altman and weighted Deming regression analyses. Flare detection sensitivity and specificity were compared between ‘minimal’ baseline and 6 week sample CRP data and the ‘continuous’ weekly CRP data. Baseline DBS ELISA assay CRP measures yielded a mean positive bias of 2.693 ± 8.640 (95% limits of agreement − 14.24 to 19.63%), when compared to reference assay data. Deming regression revealed good agreement between the DBS ELISA method and reference assay data, with baseline data slope of 0.978 and intercept -0.153. The specificity of ‘continuous’ area under the curve (AUC) CRP data (72.7%) to identify flares, was greater than ‘minimal’ AUC CRP data (54.5%). This study indicates reasonable agreement between DBS and the reference method, especially at low to mid-range CRP values. Importantly, longitudinal CRP measurement in RA patients helps to clearly identify flare and thus could assist in remote monitoring strategies and may facilitate timely therapeutic intervention.Trial registration: The Remote Arthritis Disease Activity MonitoR (RADAR) study was registered on 22/06/2016 at ClinicalTrials.gov Identifier: NCT02809547. https://clinicaltrials.gov/ct2/show/NCT02809547.

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Enhancement of Biotinidase activity in dried blood spot by Disulfide Reducing Reagent and Comparative Evaluation with Reference Method

Asian Journal of Research in Chemistry ◽

10.5958/0974-4150.2019.00069.5 ◽

2019 ◽

Vol 12 (6) ◽

pp. 366

Author(s):

Shrimant N Panaskar ◽

Dinesh Yadav ◽

Susheel K Singh ◽

Mahesh H Hampe ◽

Akshata Shivalkar

Keyword(s):

Comparative Evaluation ◽

Reference Method ◽

Dried Blood Spot ◽

Biotinidase Activity ◽

Blood Spot

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A LAMP-SNP Assay Detecting C580Y Mutation in Pfkelch13 Gene from Clinically Dried Blood Spot Samples

The Korean Journal of Parasitology ◽

10.3347/kjp.2021.59.1.15 ◽

2021 ◽

Vol 59 (1) ◽

pp. 15-22

Author(s):

Thunchanok Khammanee ◽

Nongyao Sawangjaroen ◽

Hansuk Buncherd ◽

Aung Win Tun ◽

Supinya Thanapongpichat

Keyword(s):

Field Study ◽

Artemisinin Resistance ◽

Plasmodium Species ◽

Dried Blood Spot ◽

Clinical Samples ◽

Simple Method ◽

Infected Blood ◽

And Storage ◽

C580y Mutation ◽

Blood Spot

Artemisinin resistance (ART) has been confirmed in Greater Mekong Sub-region countries. Currently, C580Y mutation on Pfkelch13 gene is known as the molecular marker for the detection of ART. Rapid and accurate detection of ART in field study is essential to guide malaria containment and elimination interventions. A simple method for collection of malaria-infected blood is to spot the blood on filter paper and is fast and easy for transportation and storage in the field study. This study aims to evaluate LAMP-SNP assay for C580Y mutation detection by introducing an extra mismatched nucleotide at the 3’ end of the FIP primer. The LAMP-SNP assay was performed in a water bath held at a temperature of 56°C for 45 min. LAMP-SNP products were interpreted by both gel-electrophoresis and HNB-visualized changes in color. The method was then tested with 120 P. falciparum DNA from dried blood spot samples. In comparing the LAMP-SNP assay results with those from DNA sequencing of the clinical samples, the 2 results fully agreed to detect C580Y. The sensitivity and specificity of the LAMP-SNP assay showed 100%. There were no cross-reactions with other Plasmodium species and other Pfkelch13 mutations. The LAMP-SNP assay performed in this study was rapid, reliable, and useful in detecting artemisinin resistance in the field study.

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Validation and Clinical Evaluation of a Novel Method To Measure Miltefosine in Leishmaniasis Patients Using Dried Blood Spot Sample Collection

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.02976-15 ◽

2016 ◽

Vol 60 (4) ◽

pp. 2081-2089 ◽

Cited By ~ 11

Author(s):

A. E. Kip ◽

H. Rosing ◽

M. J. X. Hillebrand ◽

S. Blesson ◽

B. Mengesha ◽

...

Keyword(s):

Venous Blood ◽

Dried Blood Spot ◽

Patient Specific ◽

Pharmacokinetic Studies ◽

Sample Collection ◽

Limit Of Quantification ◽

Simple Method ◽

Linear Effect ◽

Combination Treatments ◽

Blood Spot

ABSTRACTTo facilitate future pharmacokinetic studies of combination treatments against leishmaniasis in remote regions in which the disease is endemic, a simple cheap sampling method is required for miltefosine quantification. The aims of this study were to validate a liquid chromatography-tandem mass spectrometry method to quantify miltefosine in dried blood spot (DBS) samples and to validate its use with Ethiopian patients with visceral leishmaniasis (VL). Since hematocrit (Ht) levels are typically severely decreased in VL patients, returning to normal during treatment, the method was evaluated over a range of clinically relevant Ht values. Miltefosine was extracted from DBS samples using a simple method of pretreatment with methanol, resulting in >97% recovery. The method was validated over a calibration range of 10 to 2,000 ng/ml, and accuracy and precision were within ±11.2% and ≤7.0% (≤19.1% at the lower limit of quantification), respectively. The method was accurate and precise for blood spot volumes between 10 and 30 μl and for Ht levels of 20 to 35%, although a linear effect of Ht levels on miltefosine quantification was observed in the bioanalytical validation. DBS samples were stable for at least 162 days at 37°C. Clinical validation of the method using paired DBS and plasma samples from 16 VL patients showed a median observed DBS/plasma miltefosine concentration ratio of 0.99, with good correlation (Pearson'sr= 0.946). Correcting for patient-specific Ht levels did not further improve the concordance between the sampling methods. This successfully validated method to quantify miltefosine in DBS samples was demonstrated to be a valid and practical alternative to venous blood sampling that can be applied in future miltefosine pharmacokinetic studies with leishmaniasis patients, without Ht correction.

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A Simple Method to Measure Renal Function in Swine by the Plasma Clearance of Iohexol

10.20944/preprints201711.0099.v1 ◽

2017 ◽

Author(s):

Sergio Luis-Lima ◽

Consolación Garcia-Contreras ◽

Marta Vazquez-Gomez ◽

Susana Astiz ◽

Fabiola Carrara ◽

...

Keyword(s):

Plasma Clearance ◽

Quadratic Equation ◽

Concordance Correlation ◽

Limits Of Agreement ◽

Simple Method ◽

First Order ◽

Order Polynomial ◽

Experimental Trials ◽

Conscious Animals ◽

Testing Group

There is no simple method to measure glomerular filtration rate (GFR) in swine, an established model to study renal disease. We developed a protocol to measure GFR in conscious swine with the plasma clearance of iohexol. We used two groups: testing and validation, of 8 animals each. Ten milliliters of iohexol (6.47 g) were injected by the marginal auricular vein and blood samples (3 ml) were collected from the orbital sinus at different points after injection. GFR was determined considering two models: two-compartments (CL2: all samples) and one-compartment (CL1: the last six samples). In the testing group, CL1 overestimated CL2 by ~30%: CL2=245±93 and CL1=308±123 ml/mn. This error was corrected by a first order polynomial quadratic equation to CL1, which was considered the simplified method: SM=-47.909+(1.176xCL1)–(0.00063968xCL12). SM showed narrow limits of agreement with CL2, and a concordance correlation of 0.97 and a total deviation index of 14.73%. Similar results were obtained for the validation group. This protocol is reliable, reproducible, can be performed in conscious animals, uses a single dose of the marker, and requires a reduced number of samples avoiding urine collection. Finally, it portends a significant improvement in animal-welfare conditions and handling necessities in experimental trials.

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