Finger Stick Blood Collection Research Articles

Energetics and lifestyle in inherited syndromes (ELLIES Project): Feasibility of remote specimen collection for biomarker analysis.

e22545 Background: Hereditary breast cancer (BC) impacts ~39,000 individuals annually and about 10% of BCs result from an inherited mutation. Obesity is independently associated with an increased risk of BC and there is interest in studying modifiable risk factors such as nutrition, obesity, and physical activity in correlation to risk, but there is a paucity of data on the impact of lifestyle on risk and less is known about biomarkers specific to lifestyle interventions and mutation carriers. Biologic pathways are implicated in the association of obesity and cancer risk and several biomarkers have been adequately measured in fingerstick samples in controlled settings but not studied in remote specimens. To characterize metabolic risk factors and lifestyle habits over the lifetime and inform a future intervention, we explored the feasibility of obtaining sufficient specimens for biomarker analysis using home collection kits within a cohort of hereditary cancer mutation carriers. Methods: Cohort 1, recruited from a prospective registry of mutation carriers were mailed a home biospecimen collection kit containing a bloodspot collection card (BCC), a lancet for fingerstick blood collection, a tube containing a nucleic acid stabilizing buffer for saliva collection, return postage, and detailed instructions. Cohort 2, recruited from a High-Risk BC Clinic, provided a fasting intravenous blood draw and fingerstick for a BCC. Dried BCCs were analyzed for biomarkers: leptin, IGF-1, IGF binding proteins 1 and 2, glucose, and CRP. Variances between biomarker levels in home kits and plasma samples were examined. Saliva samples were evaluated for miRNA and SNPs. Results: 40 individuals were invited to participate in Cohort 1; 20 enrolled within 7 days and were mailed a kit; ~14 days occurred between mailing the kits and receiving specimens; 5 participants (25%) were unable to fill all circles on the BCCs, 1 (5%) was unable to draw blood with the lancet provided; 5 participants (25%) required additional postage and 5 (25%) were lost to follow-up. Rate of return of completed biomarker kits was 70% (15), with only one incomplete. 11 clinic patients were invited to participate in Cohort 2; 10 enrolled and provided a fasting intravenous blood draw and fingerstick for a BCC. Pearson coefficients between plasma samples and matching dried BCC samples indicated good correlations for most markers (leptin r = 0.97, p = 0.001; IGFBP1 r = 0.98, p = 0.001; IGFBP2 r = 0.97 p = 0.001; glucose r = 0.97, p = 0.003; CRP r = 0.98, p = 0.001; IGF1 r = 0.39, p = 0.30. Of the remote saliva collections, 14 of 15 samples had adequate quality and quantity of gDNA for SNP analysis and miRNA for miRNAseq. Conclusions: Remote self-collection of capillary blood and saliva was well-accepted in a cohort of hereditary mutation carriers and feasibility was established as specimens were adequate for measuring defined biomarkers.

Technical assessment of different extraction methods and transcriptome profiling of RNA isolated from small volumes of blood

Transcriptome profiling of human whole blood is used to discover biomarkers of diseases and to assess phenotypic traits. Recently, finger-stick blood collection systems have allowed a less invasive and quicker collection of peripheral blood. Such non-invasive sampling of small volumes of blood offers practical advantages. The quality of gene expression data is strictly dependent on the steps used for the sample collection, extraction, preparation and sequencing. Here we have: (i) compared the manual and automated RNA extraction of small volumes of blood using the Tempus Spin RNA isolation kit and the MagMAX for Stabilized Blood RNA Isolation kit , respectively; and (ii) assessed the effect of TURBO DNA Free treatment on the transcriptomic data of RNA isolated from small volumes of blood. We have used the QuantSeq 3′ FWD mRNA-Seq Library Prep kit to prepare RNA-seq libraries, which were sequenced on the Illumina NextSeq 500 system. The samples isolated manually displayed a higher variability in the transcriptomic data as compared to the other samples. The TURBO DNA Free treatment affected the RNA samples negatively, decreasing the RNA yield and reducing the quality and reproducibility of the transcriptomic data. We conclude that automated extraction systems should be preferred over manual extraction systems for data consistency, and that the TURBO DNA Free treatment should be avoided when working on RNA samples isolated manually from small volumes of blood.

231-LB: Innovative Blood Sampling Technology for Convenient, Nearly Painless, and Accurate Monitoring of Diabetes

Monitoring HbA1c levels in individuals affected by diabetes remains the standard of care for ensuring proper management of the condition. Typically, HbA1c is measured by venipuncture or fingerstick blood collection methods at regular intervals, and for nearly all diabetic patients, monitoring HbA1c is a lifelong endeavor. In general, venipuncture and fingerstick are standard blood collection procedures, which are considered inconvenient and painful. Patient surveys have shown that venipuncture induces fear (needle phobia) in more than 20% of adults and in 60% of children. We have designed, developed, and validated an innovative blood sampling device to collect and stabilize capillary blood from the patient’s upper arm. Using small skin incisions and vacuum, the device draws and deposits the blood onto self-contained matrices. The matrices’ cartridge is then shipped at room temperature to a clinical laboratory for HbA1c measurement. We conducted a clinical study at three sites within the U.S. to validate the use of the device on 146 enrolled participants. The primary study objective was to compare HbA1c measurement in samples collected by the device and venipuncture. Pain measurements were conducted to assess the discomfort associated with the use of the device, in comparison to venipuncture and fingerstick. The data demonstrated a strong correlation between the device and venipuncture (y=1.01-0.0131, R2=0.997) over a range of HbA1c values between 4.91 and 13.34%. Device lot-to-lot and operator-to-operator reproducibility studies showed CVs of 0.7% and 0.8%, respectively. Assessment of discomfort showed an average pain score of 1 for the device (on a scale 0 to 10) and the device was preferred over venipuncture and fingerstick by 79% of the subjects. We clinically validated an innovative blood sampling technology that provides a convenient, nearly painless, safe and effective alternative to venipuncture and fingerstick for HbA1c measurement in diabetic individuals. Disclosure E. Romeo: Employee; Self; Drawbridge Health, Inc. P.K. Bryant-Greenwood: None. K.N. Castorino: Consultant; Self; Abbott. Research Support; Self; Abbott, Dexcom, Inc., Drawbridge, Novo Nordisk Inc., Sanofi US. D.R. Liljenquist: None. B.L. Jordan: Employee; Self; Drawbridge Health. Employee; Spouse/Partner; Innovation Endeavors, LLC. M. Harrington: Employee; Self; Drawbridge Health. T. Holscher: Employee; Self; Drawbridge Health. C. Alexander: Consultant; Self; CAlexanderConsulting, LLC. V. Dokukin: Employee; Self; Drawbridge Health. M. Nerlekar: Employee; Self; Drawbridge Health. L. Williamson: Other Relationship; Self; Drawbridge Health. J. Sloane: Other Relationship; Self; Drawbridge Health.

Identifying Alpha-1 Antitrypsin Deficiency Based on Computed Tomography Evidence of Emphysema.

IntroductionChronic obstructive pulmonary disease (COPD) is most commonly caused by smoking tobacco or cigarettes. However, alpha-1 antitrypsin deficiency (AATD) is the only genetic disorder known to cause COPD and these patients often present with emphysema earlier in life and with more severe disease. Additionally, AATD patients are often misdiagnosed with other lung disorders, and the diagnosis is often delayed for up to a decade. Furthermore, several clinicians may see the patient before genetic testing is performed and an official diagnosis is made. We hypothesized that patients with radiographic emphysema on computed tomography (CT) scan of the chest would represent an enriched population of patients with a higher prevalence of alpha-1 antitrypsin (AAT) carrier or heterozygous state.MethodsWe evaluated 250 in-patients with chest computed tomography (CT) findings of emphysema, and per clinical guidelines, all were tested for AAT with Alphakit finger stick blood collection kits. Sampling 250 patients provided power to detect a carrier prevalence of 20% +/- 1.0%.ResultsA total of 250 patients were recruited of which 53% were male, 91% Caucasian, 7% African American, and 16% active smokers. They smoked an average of 39 packs per year. The prevalence of carrier status (Pi*MS or Pi*MZ) was 6.8% (95% CI (4%, 11%)). The mean forced expiratory volume in one second (FEV-1) was 53%, predicted among Pi*MM patients (n=126) and not significantly different from the Pi*MS group (50%, n=13). 69% of Pi*MM were diagnosed with asthma or COPD, vs. 79% of Pi*MS (n=14) and 100% Pi*MZ (n=3), but the difference was not significant (p=0.4).ConclusionIn the population studied, compared to a cohort of patients with abnormal pulmonary function tests (PFTs), radiographically evident emphysema did not identify patients at higher risk of being heterozygous or homozygous for AAT deficiency.

The feasibility of fingerstick blood collection for point-of-care HIV-1 viral load monitoring in rural Zambia.

Viral load monitoring for HIV treatment is recommended but not feasible in many settings. A point-of-care test using capillary blood would increase access but may require up to 200 μL of blood to achieve a lower limit of detection of 1000 copies/mL. This cross-sectional study evaluated the feasibility of collecting 200 μL of capillary blood and blood collection preferences among adults in rural Zambia. Adults seeking HIV counseling and testing at Macha Hospital were recruited in 2015. Capillary blood was collected in four 50 μL tubes. Blood collection was categorized as complete (200 μL collected), partial (all tubes filled but <200 μL obtained due to collection techniques), or incomplete (1-4 tubes attempted; <200 μL obtained due to insufficient blood flow). One fingerstick was required for 90% of the 201 participants. A median blood volume of 196 μL was collected. Complete, partial and incomplete collection was achieved in 34%, 59% and 6% of participants. The majority of participants (95%) preferred fingerstick over venous blood collection. A point-of-care viral load test requiring up to 200 μL of blood is feasible in a rural setting but would require training and supervision to ensure that sufficient blood was collected.

Evaluation of Specimen Types for Pima CD4 Point-of-Care Testing: Advantages of Fingerstick Blood Collection into an EDTA Microtube.

IntroductionEffective point-of-care testing (POCT) is reliant on optimal specimen collection, quality assured testing, and expedited return of results. Many of the POCT are designed to be used with fingerstick capillary blood to simplify the blood collection burden. However, fingerstick blood collection has inherent errors in sampling. An evaluation of the use of capillary and venous blood with CD4 POCT was conducted.MethodsThree different specimen collection methods were evaluated for compatibility using the Alere Pima CD4 assay at 5 HIV/AIDS healthcare sites in Dar es Salaam, Tanzania. At each site, whole blood specimens were collected from enrolled patients by venipuncture and fingerstick. Pima CD4 testing was performed at site of collection on venipuncture specimens (Venous) and fingerstick blood directly applied to a Pima CD4 cartridge (Capillary-Direct) and collected into an EDTA microtube (Capillary-Microtube). Venous blood was also tested at the laboratory by the reference CD4 method and Pima for comparison analysis.ResultsAll three specimen collection methods were successfully collected by healthcare workers for use with the Pima CD4 assay. When compared to the reference CD4 method, Pima CD4 testing with the Capillary-Microtube method performed similarly to Venous, while Pima CD4 counts with the Capillary-Direct method were slightly more biased (-20 cells/μL) and variable (-229 to +189 cells/μL limit of agreement). Even though all three collection methods had similar invalid Pima testing rates (10.5%, 9.8%, and 8.3% for Capillary-Direct, Capillary-Microtube, and Venous respectively), the ability to perform repeat testing with Capillary-Microtube and Venous specimens increased the likelihood of acquiring a valid CD4 result with the Pima assay.ConclusionsCapillary blood, either directly applied to Pima CD4 cartridges or collected in an EDTA microtube, and venous blood are suitable specimens for Pima CD4 testing. The advantages of capillary blood collection in an EDTA microtube are that it uses fingerstick collection which mimics venous blood and allows extra testing without additional blood collection.

Expanding the Utility of High-Sensitivity Dried Blood Spot Immunoassay Testing with Single Molecule Counting.

Dried blood spot (DBS) testing has been used for years in newborn screening and for other applications when obtaining blood by venipuncture is impractical or expensive. However, several technical challenges have restricted the use of DBS testing to qualitative assays or to analytes that are present in relatively high concentrations. The application of high-sensitivity detection using single molecule counting (SMC™) technology can potentially overcome the limitations of DBS as specimen source. A method was developed for reproducibly collecting, storing, and subsequently reconstituting DBS samples to be used with assays based on the SMC technology. Before extraction, DBS samples were scanned, and the blood spot area was calculated to normalize for sample volume and spot variability. DBS sample extraction was done using an efficient high-salt extraction buffer. DBS samples were tested using SMC-based cardiac troponin I (cTnI), prostate-specific antigen (PSA), and C-reactive protein (CRP) assays. The SMC-DBS assays showed reproducible sensitivity, precision, and the stability required for quantifying low-abundance biomarkers. These assays were not significantly impacted by normal variations in hematocrit or sample collection technique. Correlation coefficients obtained from method comparisons between SMC-DBS and laboratory-developed tests or Food and Drug Administration-cleared tests using traditional sample types were 1.08, 1.04, and 0.99 for cTnI, PSA, and high-sensitivity CRP, respectively. Combining DBS finger-stick blood collection with next-generation immunoassay technology will aid the expansion of DBS testing to protein biomarkers that are in low abundance or to low-volume samples, and will enable the development and adoption of DBS testing to far-reaching applications.

Finger stick blood collection for gene expression profiling and storage of tempus blood RNA tubes

With this report we aim to make available a standard operating procedure (SOP) developed for RNA stabilization of small blood volumes collected via a finger stick. The anticipation that this procedure may be improved through peer-review and/or readers public comments is another element motivating the publication of this SOP. Procuring blood samples from human subjects can, among other uses, enable assessment of the immune status of an individual subject via the profiling of RNA abundance using technologies such as real time PCR, NanoString, microarrays or RNA-sequencing. It is often desirable to minimize blood volumes and employ methods that are the least invasive and can be practically implemented outside of clinical settings. Finger stick blood samples are increasingly used for measurement of levels of pharmacological drugs and biological analytes. It is a simple and convenient procedure amenable for instance to field use or self-collection at home using a blood sample collection kit. Such methodologies should also enable the procurement of blood samples at high frequency for health or disease monitoring applications.

Finger stick blood collection for gene expression profiling and storage of tempus blood RNA tubes

With this report we aim to make available a standard operating procedure (SOP) developed for RNA stabilization of small blood volumes collected via a finger stick. The anticipation that this procedure may be improved through peer-review and/or readers public comments is another element motivating the publication of this SOP. Procuring blood samples from human subjects can, among other uses, enable assessment of the immune status of an individual subject via the profiling of RNA abundance using technologies such as real time PCR, NanoString, microarrays or RNA-sequencing. It is often desirable to minimize blood volumes and employ methods that are the least invasive and can be practically implemented outside of clinical settings. Finger stick blood samples are increasingly used for measurement of levels of pharmacological drugs and biological analytes. It is a simple and convenient procedure amenable for instance to field use or self-collection at home using a blood sample collection kit. Such methodologies should also enable the procurement of blood samples at high frequency for health or disease monitoring applications.

Finger stick blood collection for gene expression profiling and storage of tempus blood RNA tubes.

With this report we aim to make available a standard operating procedure (SOP) developed for RNA stabilization of small blood volumes collected via a finger stick. The anticipation that this procedure may be improved through peer-review and/or readers public comments is another element motivating the publication of this SOP. Procuring blood samples from human subjects can, among other uses, enable assessment of the immune status of an individual subject via the profiling of RNA abundance using technologies such as real time PCR, NanoString, microarrays or RNA-sequencing. It is often desirable to minimize blood volumes and employ methods that are the least invasive and can be practically implemented outside of clinical settings. Finger-stick blood samples are increasingly used for measurement of levels of pharmacological drugs and biological analytes. It is a simple and convenient procedure amenable for instance to field use or self-collection at home using a blood sample collection kit. Such methodologies should also enable the procurement of blood samples at high frequency for health or disease monitoring applications.

An investigation of fingerstick blood collection for point-of-care HIV-1 viral load monitoring in South Africa.

Viral load (VL) quantification is an important tool in determining newly developed drug resistance or problems with adherence to antiretroviral therapy (ART) in HIV-positive patients. VL monitoring is becoming the standard of care in many resource-limited settings. Testing in resource-limited settings may require sampling by fingerstick because of general shortages of skilled phlebotomists and the expense of venepuncture supplies and problems with their distribution. To assess the feasibility and ease of collecting 150 µL capillary blood needed for the use of a novel collection device following a classic fingerstick puncture. Patients were recruited by the study nurse upon arrival for routine ART monitoring at the Themba Lethu Clinic in Johannesburg, South Africa. Each step of the fingerstick and blood collection protocol was observed, and their completion or omission was recorded. One hundred and three patients consented to the study, of whom three were excluded owing to the presence of callouses. From a total of 100 patients who consented and were enrolled, 98% of collection attempts were successful and 86% of participants required only one fingerstick to successfully collect 150 µL capillary blood. Study nurse adherence to the fingerstick protocol revealed omissions in several steps that may lower the success rate of capillary blood collection and reduce the performance of a subsequent VL assay. The findings of this study support the feasibility of collecting 150 µL of capillary blood via fingerstick for point-of-care HIV-1 VL testing in a resource-limited setting.

P1-S6.35 One finger stick, two tests: integrating syphilis and HIV Rapid tests for antenatal care and reproductive health services

Background Mother-to-Child Transmission of HIV and Congenital Syphilis are two important public health problems in many developing countries. In Peru, coverage of syphilis screening is suboptimal, test results can take weeks and a high proportion of women screened are not treated, much less their sex partners. The objective of this study was to demonstrate the feasibility of implementing syphilis and HIV rapid tests at antenatal care and reproductive health services and to measure its effect on coverage of maternal syphilis screening and treatment coverage. Methods We trained midwives from 16 health centers, to offer syphilis rapid tests (Syphilis 3.0 BioLine) to women 16 to 55 years old receiving antenatal care (ANC) and seen at the delivery or miscarriage services, and to integrate it with HIV rapid tests already in place at the health network, but which were until then done from venous blood by laboratory technicians and with results given in 15 to 30 days. Results 4497 women were screened, from January to November 2010. Prevalence of syphilis (RPR reactive, TPPA(+)) was 0.9% in ANC, 0.7% in delivery services and 1.9% at the miscarriage services. The prevalence of HIV by rapid test was 0.3%. The overall coverage for syphilis and HIV rapid tests was 93% during the study, compared to the 35% coverage of RPR testing reported for 2009 (Abstract P1-S6.35 figure 1). Previous coverage of HIV testing was not documented. Test results were given to 100% of women within 20 min of finger stick blood collection during the first contact with health providers, compared to a 27 days delay found in 2009. Treatment rates for syphilis also improved from 51% in 2009 to 93% in 2010. Whereas previously there was no system in place for registering partner treatment, our data shows that 59% of partners of positive women received treatment for syphilis during the study period. Key factors in the success of implementation in Callao-Peru; were ongoing training, monitoring, and supervision of health providers who performed the rapid tests. Conclusions Implementation of syphilis rapid test proved feasible, acceptable and effective in improving screening and treatment coverage and can serve as a powerful catalyst for improvements in quality of care.

Whole genome transcript profiling from fingerstick blood samples: a comparison and feasibility study

BackgroundWhole genome gene expression profiling has revolutionized research in the past decade especially with the advent of microarrays. Recently, there have been significant improvements in whole blood RNA isolation techniques which, through stabilization of RNA at the time of sample collection, avoid bias and artifacts introduced during sample handling. Despite these improvements, current human whole blood RNA stabilization/isolation kits are limited by the requirement of a venous blood sample of at least 2.5 mL. While fingerstick blood collection has been used for many different assays, there has yet to be a kit developed to isolate high quality RNA for use in gene expression studies from such small human samples. The clinical and field testing advantages of obtaining reliable and reproducible gene expression data from a fingerstick are many; it is less invasive, time saving, more mobile, and eliminates the need of a trained phlebotomist. Furthermore, this method could also be employed in small animal studies, i.e. mice, where larger sample collections often require sacrificing the animal. In this study, we offer a rapid and simple method to extract sufficient amounts of high quality total RNA from approximately 70 μl of whole blood collected via a fingerstick using a modified protocol of the commercially available Qiagen PAXgene RNA Blood Kit.ResultsFrom two sets of fingerstick collections, about 70 uL whole blood collected via finger lancet and capillary tube, we recovered an average of 252.6 ng total RNA with an average RIN of 9.3. The post-amplification yields for 50 ng of total RNA averaged at 7.0 ug cDNA. The cDNA hybridized to Affymetrix HG-U133 Plus 2.0 GeneChips had an average % Present call of 52.5%. Both fingerstick collections were highly correlated with r2 values ranging from 0.94 to 0.97. Similarly both fingerstick collections were highly correlated to the venous collection with r2 values ranging from 0.88 to 0.96 for fingerstick collection 1 and 0.94 to 0.96 for fingerstick collection 2.ConclusionsOur comparisons of RNA quality and gene expression data of the fingerstick method with traditionally processed sample workflows demonstrate excellent RNA quality from the capillary collection as well as very high correlations of gene expression data.

EXTRACTION AND QUANTIFICATION OF CYCLOSPORINE FROM WHOLE BLOOD SAMPLES COLLECTED WITH THE CYCLOSTAT??? COLLECTION DEVICE

675 The collection of whole blood samples from a fingerstick for cyclosporine(CsA) determination could facilitate patient monitoring, allowing at home sample collection. Purpose: To evaluate the fingerstick blood collection device, CycloStat, using frozen venous whole blood samples as a model for fingerstick at home sampling. Method: A total of ninety serial whole blood samples from ten different patients were collected and stored at -80°C. The samples were thawed and spotted on the CycloStat device to compare CsA quantification in liquid and CycloStat samples. Each sample was tested a total of eight times, twice in a single assay for liquid determination, and six times in 3 assays for CycloStat sample determination. All testing was conducted with the IncStar RIA. A proprietary technology was used for CycloStat extractions, manufacturer's instructions were followed for liquid whole blood testing. Additional testing on 50 samples was conducted to determine liquid sample reproducibility. Samples with a CsA value of less than 50 ng/mL were reported as zero. Results: The intra-assay, inter-assay and overall coefficient of variation for the liquid samples were 6%, 8% and 10% respectively; for CycloStat samples the values were 4%, 9% and 9% respectively. A linear regression was performed on values from the liquid vs. the CycloStat samples. Y=1.5× + 8.7 r = 0.99, n = 90.FigureAlthough the values obtained for the spotted samples are lower than those of the liquid, the 0.99 correlation enables a simple algorithm (from the regression data) to be applied to additional CycloStat samples to obtain a correct CsA value. This simple algorithm has been found to be consistent within a sample type (in this case, frozen venous whole blood) for different patients with different lots of CycloStat and assay material. Inconclusion, these studies demonstrate the feasibility of CsA determination from a dried spot of whole blood. This technique could be used for fingerstick home sample collection for transplant patients.