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Clinical Biochemistry Jan 2020Dried specimens have been proposed in multiple environments to minimize costs associated with specimen storage and shipping in clinical studies. This report describes...
BACKGROUND
Dried specimens have been proposed in multiple environments to minimize costs associated with specimen storage and shipping in clinical studies. This report describes the development and validation of an automated method for qualitative toxicology screening of dried urine samples using LC-MS/MS.
METHODS
Urine standards containing 41 compounds were prepared and applied to filter paper cards. Dried urine was eluted from the cards using a Dried Blood Spot (DBS) autosampler from Spark Holland, which was plumbed inline with a Thermo Scientific Turboflow chromatography system for subsequent MS/MS detection with selected reaction monitoring. Limits of detection, precision of peak areas, repeatability, and carryover studies were conducted. Concordance with a reference LC-MS/MS method using liquid samples was evaluated using remnant discarded specimens.
RESULTS
The limit of detection ranged from 5 to 75 ng/mL for most compounds. At the LOD for each analyte, the peak area precision ranged from 8 to 29%. For 20 repeat injections of samples spiked at ±25% of the LOD, there was a 4% false positive rate for the 75% × LOD samples, and a 0.4% false negative rate for the +125% × LOD samples. In comparing 40 known positive specimens analyzed with the DUS method and a liquid urine reference method, there was 88% agreement. Analysis of 10 known negative specimens yielded negative results. There was no significant carryover detected up to 2000 ng/mL for any of the analytes in the assay.
CONCLUSION
Using a robotic DUS sampling an inline HTLC-MS/MS system, we have developed and validated a fully-automated and robust method for multi-analyte detection of drugs of abuse in dried urine specimens.
Topics: Chromatography, Liquid; High-Throughput Screening Assays; Humans; Limit of Detection; Robotics; Substance Abuse Detection; Substance-Related Disorders; Tandem Mass Spectrometry; Urinalysis
PubMed: 31707014
DOI: 10.1016/j.clinbiochem.2019.10.009 -
International Journal of Biomaterials 2021Amnion grafts can be preserved as freeze-dried amnion membrane (FD-AM) and amnion sponge. Preserved grafts require to be sterilized by gamma irradiation. However, each...
BACKGROUND
Amnion grafts can be preserved as freeze-dried amnion membrane (FD-AM) and amnion sponge. Preserved grafts require to be sterilized by gamma irradiation. However, each step of the process could affect its biological properties. Even so, there are only a few studies that report the influence of the preservation method and gamma irradiation on growth factor levels in preserved amniotic grafts.
METHODS
This was an experimental study with a pretest-posttest group design using a consecutive sampling technique in one batch of amnion donors at a particular time. The amnion was made into FD-AM and amnion sponge preparations, and they were sterilized with gamma irradiation (15 kGy and 25 kGy). Nonirradiated specimens served as controls, and 20 mg of each specimen was pulverized to evaluate the growth factors levels using ELISA.
RESULTS
There were significant decreases in amnion sponge compared to the FD-AM, both in transforming growth factor beta (TGF-) and basic fibroblast growth factor (bFGF) levels and in the preirradiated and 25 kGy postirradiated preparations ( ≤ 0.05). The growth factor levels in the preirradiated and postirradiated FD-AM (both 15 kGy and 25 kGy) showed significant differences ( ≤ 0.05). Likewise, the preirradiated amnion sponge group's growth factor levels compared with the postirradiated amnion sponge group also showed a significant decrease ( ≤ 0.05).
CONCLUSION
TGF- and bFGF levels were lower in amnion sponge than FD-AM. The FD-AM and amnion sponge preparations' growth factors levels were reduced following gamma irradiation sterilization. Although the decrease in growth factor levels is significant, the number of growth factor levels is still sufficient for tissue healing.
PubMed: 33927767
DOI: 10.1155/2021/6685225 -
Zhejiang Da Xue Xue Bao. Yi Xue Ban =... Oct 2020To explore effects of different delivery and storage conditions on concentrations of amino acids and carnitines in neonatal dried blood spots (DBS), so as to provide...
OBJECTIVE
To explore effects of different delivery and storage conditions on concentrations of amino acids and carnitines in neonatal dried blood spots (DBS), so as to provide evidence for improving accurate and reliable detection by tandem mass spectrometry.
METHODS
A total of 1 254 616 newborn DBS samples in Newborn Screening Center of Zhejiang Province were delivered and stored at room temperature (group A, =338 467), delivered by cold-chain logistics system and stored at low temperature (group B, =480 021), or delivered by cold-chain logistics system and stored at low temperature and low humidity (group C, = 436 128), respectively. The concentrations of amino acids and carnitines in DBS were detected by tandem mass spectrometry. Data analysis was performed by SPSS 24.0 to explore the influence of temperature and humidity on the concentrations of amino acids and carnitines.
RESULTS
The concentrations of amino acids and carnitines in the three groups were skewed, and the differences in amino acid and carnitine concentrations among groups were statistically significant (all <0.01). The median concentration of tyrosine was lower in group A than those in group B and group C by 18%and 16%respectively, while there was no significant difference between the last two groups. The median concentrations of methionine were lower in group A and group B than that in group C by 15%and 11%, respectively. The median concentrations of arginine were lower in group A and group B than that in group C by 12%and 25%, respectively. The median concentration of free carnitine (C0) was higher in group A than that in group C by 12%, while there was no significant difference between group A and group B. The median concentrations of acetylcarnitine (C2), propionyl carnitine (C3), C3DC+C4OH, C5DC+C6OH and hexadecanoyl carnitine (C16) were lower in group A than those in group B and group C by 21%-64%. The concentrations of other amino acids and acylcarnitines differed little among three groups. The monthly median coefficients of variation of other amino acids and carnitines in group A were higher than those in group B and group C except for citrulline, C4DC+C5OH and isovalerylcarnitine (C5).
CONCLUSIONS
Cold-chain logistics system and storage in low temperature and low humidity can effectively reduce degradation of some amino acids and carnitines in DBS, improve the accuracy and reliability of detection, and thus ensures the quality of screening for neonatal metabolic diseases.
Topics: Amino Acids; Carnitine; Dried Blood Spot Testing; Humans; Humidity; Infant, Newborn; Neonatal Screening; Reproducibility of Results; Specimen Handling; Tandem Mass Spectrometry; Temperature; Time Factors
PubMed: 33210481
DOI: 10.3785/j.issn.1008-9292.2020.10.03 -
Mass Spectrometry (Tokyo, Japan) 2022Dried blood spot (DBS) is the standard specimen for the newborn screening of inborn errors of metabolism (IEM) by tandem mass spectrometry. Availability of DBS for the...
Dried blood spot (DBS) is the standard specimen for the newborn screening of inborn errors of metabolism (IEM) by tandem mass spectrometry. Availability of DBS for the mass spectrometric analysis of the diagnostic marker proteins, transferrin (Tf) and apolipoprotein CIII (apoCIII), of congenital disorders of glycosylation (CDG) was examined. Recovery of Tf from DBS was only slightly reduced compared with fresh serum. Although oxidation of the core polypeptides was observed, glycans of Tf and apoCIII were unaffected by storage of DBS in the ambient environment for at least 1 month. The combination of DBS and the triple quadrupole mass spectrometer used for IEM screening was sufficient to characterize the aberrant glycoprofiles of Tf and apoCIII in CDG. DBS or dried serum spot on filter paper can reduce the cost of sample transportation and potentially promote mass spectrometric screening of CDG.
PubMed: 36713804
DOI: 10.5702/massspectrometry.A0113 -
Therapeutic Drug Monitoring Jun 2021With the increasing prevalence of multidrug resistant organisms, therapeutic drug monitoring (TDM) has become a common tool for assuring the safety and efficacy of... (Review)
Review
BACKGROUND
With the increasing prevalence of multidrug resistant organisms, therapeutic drug monitoring (TDM) has become a common tool for assuring the safety and efficacy of antimicrobial drugs at higher doses. Microsampling techniques, including dried blood spotting (DBS) and volumetric absorptive microsampling (VAMS), are attractive tools for TDM and pediatric clinical research. For microsampling techniques to be a useful tool for TDM, it is necessary to establish the blood-plasma correlation and the therapeutic window of antimicrobial drugs in the blood.
METHODS
DBS involves the collection of small volumes of blood (30-50 µL per spot) on a filter paper, whereas VAMS allows the accurate and precise collection of a fixed volume of blood (10-30 µL) with microsampling devices. One of the major advantages of VAMS is that it reduces or eliminates the volumetric blood hematocrit (HCT) bias associated with DBS. Liquid chromatography with tandem mass spectrometry is a powerful tool for the accurate quantification of antimicrobial drugs from small volumes of blood specimens.
RESULTS
This review summarizes the recent liquid chromatography with tandem mass spectrometry assays that have used DBS and VAMS approaches for quantifying antimicrobial drugs. Sample collection, extraction, validation outcomes, including the interassay and intra-assay accuracy and precision, recovery, stability, and matrix effect, as well as the clinical application of these assays and their potential as tools of TDM are discussed herein.
CONCLUSIONS
Microsampling techniques, such as VAMS, provide an alternative approach to traditional plasma sample collection for TDM.
Topics: Anti-Infective Agents; Blood Specimen Collection; Child; Chromatography, Liquid; Drug Monitoring; Humans; Tandem Mass Spectrometry
PubMed: 33278241
DOI: 10.1097/FTD.0000000000000845 -
JMIR Public Health and Surveillance Jun 2020The severe acute respiratory coronavirus 2 (SARS-CoV-2) pandemic calls for expanded opportunities for testing, including novel testing strategies such as home-collected...
BACKGROUND
The severe acute respiratory coronavirus 2 (SARS-CoV-2) pandemic calls for expanded opportunities for testing, including novel testing strategies such as home-collected specimens.
OBJECTIVE
We aimed to understand whether oropharyngeal swab (OPS), saliva, and dried blood spot (DBS) specimens collected by participants at home and mailed to a laboratory were sufficient for use in diagnostic and serology tests of SARS-CoV-2.
METHODS
Eligible participants consented online and were mailed a participant-collection kit to support collection of three specimens for SARS-CoV-2 testing: saliva, OPS, and DBS. Participants performed the specimen collection procedures during a telehealth video appointment while clinical observers watched and documented the suitability of the collection. The biological sufficiency of the specimens for detection of SARS-CoV-2 by reverse transcriptase-polymerase chain reaction and serology testing was assessed by laboratorians using visual inspection and quantification of the nucleic acid contents of the samples by ribonuclease P (RNase P) measurements.
RESULTS
Of the enrolled participants,153/159 (96.2%) returned their kits, which were included in this analysis. All these participants attended their video appointments. Clinical observers assessed that of the samples collected, 147/153 (96.1%) of the saliva samples, 146/151 (96.7%) of the oropharyngeal samples, and 135/145 (93.1%) of the DBS samples were of sufficient quality for submission for laboratory testing; 100% of the OPS samples and 98% of the saliva samples had cycle threshold values for RNase P <30, indicating that the samples contained sufficient nucleic acid for RNA-PCR testing for SARS-CoV-2.
CONCLUSIONS
These pilot data indicate that most participant-collected OPS, saliva, and DBS specimens are suitable and sufficient for testing for SARS-CoV-2 RNA and serology. Clinical observers rated the collection of specimens as suitable for testing, and visual and quantitative laboratory assessment indicated that the specimens were biologically sufficient. These data support the utility of participant-collected and mailed-in specimens for SARS-CoV-2 testing.
INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID)
RR2-10.2196/19054.
Topics: Adolescent; Adult; Aged; COVID-19; COVID-19 Testing; Clinical Laboratory Techniques; Cohort Studies; Coronavirus Infections; Dried Blood Spot Testing; Female; Health Services Research; Humans; Male; Middle Aged; Oropharynx; Pandemics; Pilot Projects; Pneumonia, Viral; Saliva; Specimen Handling; Telemedicine; Young Adult
PubMed: 32479412
DOI: 10.2196/19731 -
American Journal of Human Biology : the... Sep 2020This study investigates how factors related to collection, storage, transport time, and environmental conditions affect the quality and accuracy of analyses of dried...
OBJECTIVES
This study investigates how factors related to collection, storage, transport time, and environmental conditions affect the quality and accuracy of analyses of dried blood spot (DBS) samples.
METHODS
Data come from the 2016 Health and Retirement Study (HRS) DBS laboratory reports and the HRS merged with the National Climatic Data Center (NCDC) Global Historical Climate Network Daily (NCDC GHCN-Daily) and the NCDC Local Climatological Data, by zip code. We ran regression models to examine the associations between assay values based on DBS for five analytes (total cholesterol, high-density lipoprotein (HDL) cholesterol, glycosylated hemoglobin (HbA1c), C-reactive protein (CRP), and cystatin C) and the characteristics of DBS cards and drops, shipping time, and temperature, and humidity at the time of collection.
RESULTS
We found cholesterol measures to be sensitive to many factors including small spots, shipping time, high temperature and humidity. Small spots in DBS cards are related to lower values across all analytes. Longer DBS transit time before freezing is associated with lower values of total and HDL cholesterol and cystatin C. Results were similar whether or not venous blood sample values were included in equations.
CONCLUSIONS
Small spots, long shipping time, and exposure to high temperature and humidity need to be avoided if possible. Quality of spots and cards and information on shipping time and conditions should be coded with the data to make adjustments in values when necessary. The different results across analytes indicate that results cannot be generalized to all DBS assays.
Topics: Dried Blood Spot Testing; Hot Temperature; Humans; Humidity; Regression Analysis; Specimen Handling; United States
PubMed: 31922324
DOI: 10.1002/ajhb.23390 -
Metabolomics : Official Journal of the... Jul 2020Blood-based sample collection is a challenge, and dried blood spots (DBS) represent an attractive alternative. However, for DBSs to be an alternative to venous blood it...
INTRODUCTION
Blood-based sample collection is a challenge, and dried blood spots (DBS) represent an attractive alternative. However, for DBSs to be an alternative to venous blood it is important that these samples are able to deliver comparable associations with clinical outcomes. To explore this we looked to see if lipid profile data could be used to predict the concentration of triglyceride, HDL, LDL and total cholesterol in DBSs using markers identified in plasma.
OBJECTIVES
To determine if DBSs can be used as an alternative to venous blood in both research and clinical settings, and to determine if machine learning could predict 'clinical lipid' concentration from lipid profile data.
METHODS
Lipid profiles were generated from plasma (n = 777) and DBS (n = 835) samples. Random forest was applied to identify and validate panels of lipid markers in plasma, which were translated into the DBS cohort to provide robust measures of the four 'clinical lipids'.
RESULTS
In plasma samples panels of lipid markers were identified that could predict the concentration of the 'clinical lipids' with correlations between estimated and measured triglyceride, HDL, LDL and total cholesterol of 0.920, 0.743, 0.580 and 0.424 respectively. When translated into DBS samples, correlations of 0.836, 0.591, 0.561 and 0.569 were achieved for triglyceride, HDL, LDL and total cholesterol.
CONCLUSION
DBSs represent an alternative to venous blood, however further work is required to improve the combined lipidomics and machine learning approach to develop it for use in health monitoring.
Topics: Adolescent; Biomarkers; Blood Specimen Collection; Child; Cholesterol; Cholesterol, HDL; Cholesterol, LDL; Cohort Studies; Dried Blood Spot Testing; Female; Humans; Lipidomics; Lipids; Machine Learning; Male; Middle Aged; Netherlands; Triglycerides
PubMed: 32710150
DOI: 10.1007/s11306-020-01703-0 -
Journal of Acquired Immune Deficiency... Mar 2022Dried plasma spot specimens may be a viable alternative to traditional liquid plasma in field settings, but the diagnostic accuracy is not well understood. (Meta-Analysis)
Meta-Analysis
BACKGROUND
Dried plasma spot specimens may be a viable alternative to traditional liquid plasma in field settings, but the diagnostic accuracy is not well understood.
METHODS
Standard databases (PubMed and Medline), conferences, and gray literature were searched until January 2019. The quality of evidence was evaluated using the Standards for Reporting Studies of Diagnostic Accuracy and Quality Assessment of Diagnostic Accuracy Studies-2 criteria. We used univariate and bivariate random effects models to determine misclassification, sensitivity, and specificity across multiple thresholds, overall and for each viral load technology, and to account for between-study variation.
RESULTS
We identified 23 studies for inclusion in the systematic review that compared the diagnostic accuracy of dried plasma spots with that of plasma. Primary data from 16 of the 23 studies were shared and included in the meta-analysis, representing 18 countries, totaling 1847 paired dried plasma spot:plasma data points. The mean bias of dried plasma spot specimens compared with that of plasma was 0.28 log10 copies/mL, whereas the difference in median viral load was 2.25 log10 copies/mL. More dried plasma spot values were undetectable compared with plasma values (43.6% vs. 29.8%). Analyzing all technologies together, the sensitivity and specificity of dried plasma spot specimens were >92% across all treatment failure thresholds compared and total misclassification <5.4% across all treatment failure thresholds compared. Some technologies had lower sensitivity or specificity; however, the results were typically consistent across treatment failure thresholds.
DISCUSSION
Overall, dried plasma spot specimens performed relatively well compared with plasma with sensitivity and specificity values greater than 90% and misclassification rates less than 10% across all treatment failure thresholds reviewed.
Topics: Dried Blood Spot Testing; HIV Infections; HIV-1; Humans; RNA, Viral; Sensitivity and Specificity; Treatment Failure; Viral Load
PubMed: 34732684
DOI: 10.1097/QAI.0000000000002855 -
BioTechniques Apr 2021The aim of this study was to assess the DNA preservation capability of cellulose paper towel and blotting paper as low-cost alternatives to commercial DNA preservation...
The aim of this study was to assess the DNA preservation capability of cellulose paper towel and blotting paper as low-cost alternatives to commercial DNA preservation products. Chicken blood was applied as DNA source to each paper towel, blotting paper, FTA cards and DNA/RNA Shield™. All samples were stored at room temperature for 130 days. DNA extraction from dried blood spots was performed after various time periods using Tris-EDTA and NaOH protocols. PCR activity and the mean amount of DNA isolated from paper towels were reliable. The results of this study demonstrated that cellulose-based blotting paper and especially paper towel had considerable DNA binding and preservation capacity for at least 130 days at room temperature without DNA degradation.
Topics: Cellulose; DNA; Genomics; Polymerase Chain Reaction; Specimen Handling
PubMed: 33749333
DOI: 10.2144/btn-2020-0158