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Journal of Pharmaceutical and... Jun 2024Drug testing commonly use urine as a specimen and immunoassays for screening. The need for supervised urine collection has led to an interest in alternative specimens...
BACKGROUND
Drug testing commonly use urine as a specimen and immunoassays for screening. The need for supervised urine collection has led to an interest in alternative specimens and a need for using mass spectrometry methods already for screening. In addition, mass spectrometry methods allow for broad multipanel screening which of great value because of the increased number of substances that needs to be covered has increased over time. One alternative specimen of interest for drugs of abuse testing is dried blood spots (DBS) and this work aimed at developing multipanel screening methods based on selected reaction monitoring liquid chromatography - mass spectrometry for both urine and dried finger blood as specimens.
MATERIALS AND METHODS
The urine method comprised 37 analytes and utilised salted out liquid/liquid extraction in 96-well format, respectively, and the blood method comprised 35 analytes, a 10 µL volumetric DBS device and a two-step solvent extraction procedure. In both cases stable isotope labelled internal standards were used for almost all analytes.
RESULTS
The methods were validated according to forensic standard. The lowest reporting limits were generally set at 100 ng/mL for urine and 1 ng/mL for blood and the accuracy and imprecision were within limits of 15 and 20%. The methods were applied in a clinical study on patients receiving methadone maintenance treatment for opioid dependence. Methadone was detected in all urine and DBS samples, for urine sometimes below the commonly applied screening cutoff limit of 300 ng/mL. In 20 out of 99 cases no other drug was detected in any specimen. The most commonly other detected substances were pregabalin, amphetamine, alprazolam, zopiclone and THCCOOH. Findings in urine and DBS generally agreed well but more positives were detected in DBS.
CONCLUSION
Multipanel methods using liquid chromatography - mass spectrometry suitable for clinical drug screening were successfully developed for urine and blood collected by finger-pricking and stored as DBS.
Topics: Humans; Chromatography, Liquid; Liquid Chromatography-Mass Spectrometry; Tandem Mass Spectrometry; Methadone; Dried Blood Spot Testing
PubMed: 38457867
DOI: 10.1016/j.jpba.2024.116075 -
Molecular Genetics and Metabolism May 2024Phenylketonuria (PKU) requires regular phenylalanine monitoring to ensure optimal outcome. However, home sampling methods used for monitoring suffer high pre-analytical...
INTRODUCTION
Phenylketonuria (PKU) requires regular phenylalanine monitoring to ensure optimal outcome. However, home sampling methods used for monitoring suffer high pre-analytical variability, inter-laboratory variability and turn-around-times, highlighting the need for alternative methods of home sampling or monitoring.
METHODS
A survey was distributed through email and social media to (parents of) PKU patients and professionals working in inherited metabolic diseases in Denmark, The Netherlands, and United Kingdom regarding satisfaction with current home sampling methods and expectations for future point-of-care testing (POCT).
RESULTS
210 parents, 156 patients and 95 professionals completed the survey. Countries, and parents and patients were analysed together, in absence of significant group differences for most questions. Important results are: 1) Many patients take less home samples than advised. 2) The majority of (parents of) PKU patients are (somewhat) dissatisfied with their home sampling method, especially with turn-around-times (3-5 days). 3) 37% of professionals are dissatisfied with their home sampling method and 45% with the turn-around-times. 4) All responders are positive towards developments for POCT: 97% (n = 332) of (parents of) patients is willing to use a POC-device and 76% (n = 61) of professionals would recommend their patients to use a POC-device. 5) Concerns from all participants for future POC-devices are costs/reimbursements and accuracy, and to professionals specifically, accessibility to results, over-testing, patient anxiety, and patients adjusting their diet without consultation.
CONCLUSION
The PKU community is (somewhat) dissatisfied with current home sampling methods, highlighting the need for alternatives of Phe monitoring. POCT might be such an alternative and the community is eager for its arrival.
Topics: Humans; Phenylketonurias; Point-of-Care Testing; Male; Female; Surveys and Questionnaires; Parents; Blood Specimen Collection; United Kingdom; Netherlands; Adult; Patient Satisfaction; Phenylalanine; Denmark; Child; Adolescent
PubMed: 38442492
DOI: 10.1016/j.ymgme.2024.108361 -
Royal Society Open Science Feb 2024The article's main aim is to assess the mechanical behaviour of linden under high-rate loadings (impact) and its change due to changes in moisture content (MC) over...
The article's main aim is to assess the mechanical behaviour of linden under high-rate loadings (impact) and its change due to changes in moisture content (MC) over fibre saturation point. For assessing the mechanical properties of green wood, mainly the data of the dried wood is not applicable since the moisture content can drastically affect the mechanical properties of the wood. By testing both dried and high-moisture-content wood, we can understand a general viewpoint toward the effect of the moisture content on the impact behaviour of the wood. Several test samples were made of linden wood with different moisture content levels of 11%, 60% and 160%. A drop-weight impact machine tested the specimens to measure the reaction force of the hammer during a very short impact period. The results of the tests were parameters such as force-time chart, the maximum force required for crack initiation, the impact bending strength (IBS) and the work needed for crack initiation. The results indicated an increase in MC decreases the maximum force, work required for crack initiation and IBS drastically. However, when MC exceeded the fibre saturation point (FSP), there was no further influence on the force pattern and maximum required force.
PubMed: 38420630
DOI: 10.1098/rsos.231685 -
BMC Infectious Diseases Feb 2024Sexually transmitted infections (STIs) such as syphilis and HIV remain to be a significant public health issue worldwide. Dual rapid point-of-care tests (POCTs) have...
BACKGROUND
Sexually transmitted infections (STIs) such as syphilis and HIV remain to be a significant public health issue worldwide. Dual rapid point-of-care tests (POCTs) have shown promise for detecting antibodies to HIV and syphilis but have not been fully evaluated in the field. Our study supported the WHO ProSPeRo study on Sexually Transmitted Infection Point-of-Care Testing (STI POCT) by providing external quality assessment (EQA) for HIV and syphilis testing in reference laboratories and their associated clinical sites in seven countries.
METHODS
HIV/syphilis serum liquid and dried tube specimen (DTS) panels were prepared by CDC. Liquid panels were distributed to the reference laboratories for three rounds of testing using commercially and locally available laboratory-based serological tests. DTS panels were sent to the clinical testing sites for 8 rounds of POC testing using the Abbott SD BIOLINE HIV/Syphilis Duo test (hereafter referred to as SD BIOLINE) and the Chembio Dual Path Platform (DPP) HIV-Syphilis assay. EQA panels were tested at CDC using the Rapid Plasma Reagin (RPR) test and the Treponema pallidum Particle Agglutination assay (TP-PA) for syphilis antibodies. Genetic Systems HIV-1/HIV-2 Plus O EIA, Geenius HIV Supplemental Assay and the Oraquick Advance HIV test were used to detect HIV antibodies in the EQA panels. Results from the reference laboratories and POCT sites were compared to those obtained at the CDC and a percentage agreement was calculated.
RESULTS
Qualitative RPR and TP-PA performed at the reference laboratories demonstrated 95.4-100% agreement with CDC results while quantitative RPR and TP-PA tests demonstrated 87.7% and 89.2% agreement, respectively. A 93.8% concordance rate was observed for qualitative HIV testing in laboratories. EQA testing at clinical sites using dual tests showed 98.7% and 99.1% agreement for detection of HIV antibodies and eight out of 10 sites had > 95.8% agreement for syphilis testing. However, two clinical sites showed only 65.0-66.7% agreement for SD BIOLINE and 84.0-86.7% for DPP, respectively, for syphilis testing.
CONCLUSIONS
Overall, laboratories demonstrated high EQA performance in this study. Both HIV/syphilis POCTs gave expected results in the clinic-based evaluations using DTS. However, testing errors were identified in a few testing sites suggesting the necessity for continuous training and monitoring the quality of POC testing.
Topics: Humans; Syphilis; Treponema pallidum; HIV Antibodies; HIV Infections; Sensitivity and Specificity; Antibodies, Bacterial; Point-of-Care Testing; Syphilis Serodiagnosis; HIV-2; HIV-1; World Health Organization; Point-of-Care Systems
PubMed: 38418989
DOI: 10.1186/s12879-024-09027-3 -
General Dentistry 2024The aim of this study was to evaluate the influence of the type of etchant on the shear bond strength (SBS) of metallic brackets to enamel and the Adhesive Remnant Index...
The aim of this study was to evaluate the influence of the type of etchant on the shear bond strength (SBS) of metallic brackets to enamel and the Adhesive Remnant Index (ARI) after debonding. A total of 30 mandibular and maxillary premolars were randomly distributed into groups (n = 10) treated with 1 of 3 enamel surface-conditioning agents: 35% phosphoric acid (PA), 35% glycolic acid (GA), or 35% ferulic acid (FA). The designated acid was applied to the buccal enamel surface of the tooth for 20 seconds, and the tooth was then rinsed with distilled water for 20 seconds and air dried for 5 seconds. A metal bracket was bonded to the prepared surface with light-cured orthodontic resin. After 24 hours, the bracket-tooth interface was submitted to SBS testing in a universal testing machine at a speed of 0.5 mm/min. After debonding, the enamel surface was observed under a stereomicroscope (×20 magnification) to determine the ARI. The generalized linear models showed that the PA and GA groups presented significantly higher SBSs than the FA group (P = 0.0003). The ARI was significantly higher in specimens treated with PA than with the other acids (P < 0.05; Kruskal-Wallis and Dunn tests), with a larger quantity of adhesive remaining adhered to the tooth. Both PA and GA are effective for bonding brackets, but GA resulted in a lower percentage of adhesive remnant adhered to the enamel.
Topics: Humans; Orthodontic Brackets; Dental Enamel; Phosphoric Acids; Coumaric Acids; Glycolates
PubMed: 38411486
DOI: No ID Found -
Current Opinion in Lipidology Jun 2024Newborn screening is one of the most successful public health programs of the last century and offers unparalleled access to universal screening for a variety of... (Review)
Review
PURPOSE OF REVIEW
Newborn screening is one of the most successful public health programs of the last century and offers unparalleled access to universal screening for a variety of metabolic and other disorders. Interest in development of newborn screening for lipid disorders has intensified in recent years. Screening newborns for lipid disorders has important implications for the health of the newborn as well as their relatives, and in the case of more common lipid disorders like familial hypercholesterolemia, could have important public health implications.
RECENT FINDINGS
Recent studies have demonstrated feasibility of measuring biomarkers for heterozygous familial hypercholesterolemia from newborn screening dried blood spot specimens. Another lipid disorder, cerebrotendinous xanthomatosis, is currently amenable to newborn screening utilizing currently available assays. New research in next-generation sequencing as a primary screen in newborns will also identify both common and rare lipid disorders in newborns.
SUMMARY
Historically, newborn screening for lipid disorders was not done for many reasons, but new research has developed testing methods that may successfully identify common and rare lipid disorders. This will impact the health of the newborn but could also impact family members and public health.
Topics: Humans; Neonatal Screening; Infant, Newborn; Biomarkers
PubMed: 38408035
DOI: 10.1097/MOL.0000000000000928 -
Journal of Pharmaceutical and... May 2024Volumetric absorptive microsampling (VAMS) is increasingly proposed as a clinically reliable therapeutic drug monitoring (TDM) sampling methodology. The study aimed to...
Volumetric absorptive microsampling (VAMS) is increasingly proposed as a clinically reliable therapeutic drug monitoring (TDM) sampling methodology. The study aimed to establish the reliability and real-life feasibility of patient self-collected capillary VAMS for TDM of antiseizure medication (ASMs), using plasma ASMs concentrations from venous blood as a reference standard. Nurses collected venous and capillary blood samples using VAMS. Afterward, persons with epilepsy (PWE) performed VAMS sampling by themselves. All samples were analyzed by UHPLC-MS/MS. We performed a cross-validation study, comparing ASMs concentrations obtained by VAMS nurses and patients' self-collected versus plasma through Bland-Altman analysis and Passing-Bablok regression. We enrolled 301 PWE (M: F 42.5%:57.5%; mean age 44±16 years), treated with 13 ASMs, providing a total of 464 measurements. Statistical analysis comparing VAMS self-collected versus plasma ASMs concentrations showed a bias close to zero and slope and intercept values indicating a good agreement for CBZ, LCS, LEV, LTG, OXC, PB, and PHT, while a systematic difference between the two methods was found for VPA, PMP, TPM and ZNS. This is the first study showing the reliability and feasibility of the real-world application of PWE self-collected VAMS for most of the ASMs considered, giving a promising basis for at-home VAMS applications.
Topics: Humans; Adult; Middle Aged; Tandem Mass Spectrometry; Drug Monitoring; Reproducibility of Results; Feasibility Studies; Blood Specimen Collection; Dried Blood Spot Testing; Epilepsy
PubMed: 38401349
DOI: 10.1016/j.jpba.2024.116065 -
International Journal of Neonatal... Feb 2024Very-long-chain acyl-CoA dehydrogenase (VLCAD) deficiency is a long-chain fatty acid oxidation disorder that manifests as either a severe phenotype associated with...
Using the C14:1/Medium-Chain Acylcarnitine Ratio Instead of C14:1 to Reduce False-Positive Results for Very-Long-Chain Acyl-CoA Dehydrogenase Deficiency in Newborn Screening in Japan.
Very-long-chain acyl-CoA dehydrogenase (VLCAD) deficiency is a long-chain fatty acid oxidation disorder that manifests as either a severe phenotype associated with cardiomyopathy, a hypoglycemic phenotype, or a myopathic phenotype. As the hypoglycemic phenotype can cause sudden infant death, VLCAD deficiency is included in newborn screening (NBS) panels in many countries. The tetradecenoylcarnitine (C14:1) level in dried blood specimens is commonly used as a primary marker for VLCAD deficiency in NBS panels. Its ratio to acetylcarnitine (C2) and various other acylcarnitines is used as secondary markers. In Japan, tandem mass spectrometry-based NBS, initially launched as a pilot study in 1997, was introduced to the nationwide NBS program in 2013. In the present study, we evaluated levels of acylcarnitine with various chain lengths (C18 to C2), free carnitine, and their ratios in 175 infants who tested positive for VLCAD deficiency with C14:1 and C14:1/C2 ratios. Our analyses indicated that the ratios of C14:1 to medium-chain acylcarnitines (C10, C8, and C6) were the most effective markers in reducing false-positive rates. Their use with appropriate cutoffs is expected to improve NBS performance for VLCAD deficiency.
PubMed: 38390979
DOI: 10.3390/ijns10010015 -
The Journal of Applied Laboratory... May 2024In addition to newborn screening, dried blood spots (DBSs) are used for a wide variety of analytes for clinical, epidemiological, and research purposes. Guidelines on...
BACKGROUND
In addition to newborn screening, dried blood spots (DBSs) are used for a wide variety of analytes for clinical, epidemiological, and research purposes. Guidelines on DBS collection, storage, and transport are available, but it is suggested that each laboratory should establish its own acceptance criteria.
METHODS
An optical scanning device was developed to assess the quality of DBSs received in the newborn screening laboratory from 11 maternity wards between 2013 and 2018. The algorithm was adjusted to agree with the visual examination consensus of experienced laboratory personnel. Once validated, the algorithm was used to categorize DBS specimens as either proper or improper. Improper DBS specimens were further divided based on 4 types of specimen defects.
RESULTS
In total, 27 301 DBSs were analyzed. Compared with an annual DBS rejection rate of about 1%, automated scanning rejected 26.96% of the specimens as having at least one defect. The most common specimen defect was multi-spotting (ragged DBS, 19.13%). Among maternity wards, improper specimen rates varied greatly between 5.70% and 49.92%.
CONCLUSIONS
Improper specimen rates, as well as the dominant type of defect(s), are mainly institution-dependent, with various maternity wards consistently showing specific patterns of both parameters over time. Although validated in agreement with experienced laboratory personnel consensus, automated analysis rejects significantly more specimens. While continuous staff training, specimen quality monitoring, and problem-reporting to maternities is recommended, a thorough quality assessment strategy should also be implemented by every newborn screening laboratory. An important role in this regard may be played by automation in the form of optical scanning devices.
Topics: Humans; Neonatal Screening; Infant, Newborn; Dried Blood Spot Testing; Algorithms; Quality Assurance, Health Care; Quality Control; Blood Specimen Collection
PubMed: 38384160
DOI: 10.1093/jalm/jfae003 -
Journal of Medical Entomology Feb 2024Chagas disease, caused by the protozoan Trypanosoma cruzi, is a zoonosis primarily found in rural areas of Latin America. It is considered a neglected tropical disease,...
Chagas disease, caused by the protozoan Trypanosoma cruzi, is a zoonosis primarily found in rural areas of Latin America. It is considered a neglected tropical disease, and Triatoma dimidiata is the main vector of the parasite in Central America. Despite efforts, Chagas disease continues to be a public health concern, and vector control remains a primary tool to reduce transmission. In this study, we tested the hypothesis that highly abundant bacteria in the gut of T. dimidiata inhibit the growth of T. cruzi. To achieve this, bacterial diversity in the gut of T. dimidiata specimens from Costa Rica was characterized by metabarcoding of the 16S rRNA, microbial isolation was performed, and the effect of freeze-dried supernatants of the isolates on T. cruzi was investigated. Metabarcoding showed that the most abundant genera in the gut were Corynebacterium, Tsukamurella, Brevibacterium, and Staphylococcus. Barcoding and sequences comparison confirmed that 8 of the 30 most abundant amplicon sequence variants (ASVs) were isolated, and 2 of them showed an inhibitory effect on the growth of T. cruzi epimastigotes. These bacteria correspond to isolates of Tsukamurella and Brevibacterium, which were respectively the second and sixth most abundant ASVs in the gut of T. dimidiata. Notably, only the isolate of Brevibacterium showed a significant difference in growth inhibition against epimastigotes of both T. cruzi strains tested. These findings suggest that the gut microbiota of T. dimidiata may play an active role in modulating parasite development.
PubMed: 38381588
DOI: 10.1093/jme/tjae012