bluuebunny/crossref_metadata_2025_split huggingface dataset:
B-275 Retrospective Study for Mpox Detection: Validation of a Real Time PCR Kit for its Fast Diagnosis
Authors: Peris M , Milagro A , Fortuño B , Martínez-Mateos D , Gilaberte B , Dehesa-García B , García-Manrique B , Laborda L , Latorre-Millán M , Aspiroz-Sancho C 
Published: September 2023

Abstract

Background
In an increasingly globalised and interconnected world, a small outbreak of an infectious disease (such as monkeypox in 2022) can turn into a global health emergency. In this context, global health systems must develop effective strategies to interrupt transmission as early as possible by identifying cases and sources of infection. Since clinical diagnostic products must be externally validated to comply with current regulations, the aim of this collaborative and retrospective study was to evaluate the clinical sensitivity and specificity of the VIASURE Monkey pox Real time PCR detection kit (CerTest Biotec) for the detection of monkeypox by Real time PCR.

Methods
This is a retrospective and collaborative study conducted at the Microbiology Department of the Miguel Servet University Hospital at the request of the company Certest Biotec SL with the approvement of the Aragon research ethics committee (PI22/412; 5 Oct 2022). A total of 192 suspected MPX samples were collected in 2022–2023. RealStar Orthopoxvirus PCR kit 1.0, Altona Diagnostics and Sanger sequencing were used as reference techniques. Clinical sensitivity and specificity (95% CI) were calculated with MetaDisc 1.4 software, while overall concordance and quantitative correlation between both real-time PCR kits was checked with the statistical package SPSS v24 (IBM Corporation).

Results
All the results obtained by the kit under study coincided with the reference kit ones (being 69 of them positives for monkeypox). Therefore, clinical sensitivity and specificity values are 100%. Furthermore, the overall agreement was almost perfect (100, 98–100) as was the Spearman correlation test (rs = 0.78; P &lt; 0.001). All positive samples were sequenced and confirmed the presence of the virus.

Conclusion
This retrospective study demonstrates the good clinical parameters and strong overall agreement of the VIASURE® Monkey pox virus Real Time PCR detection kit (CerTest Biotec) compared to both the RealStar Orthopoxvirus PCR 1.0 kit and Sanger sequencing results. In addition, the stabilised, ready-to-use format allows storage at room temperature, and facilitates transport, as it does not need to be refrigerated. This makes it environmentally friendly and reduces additional costs.
B-298 Retrospective Analysis of Fentanyl Positivity at a Large Academic Center Reveals Trends in Illicit Fentanyl Use
Authors: Berry M , Cohen L , Manthei D , Gherasim C 
Published: September 2023

Abstract

Background
National drug-involved overdose deaths continue to rise with deaths involving synthetic opioids (primarily fentanyl) showing the most significant upward trend. Fentanyl is a synthetic opioid with short-acting analgesic activity and a 50–100 times higher potency compared to morphine. Pharmaceutical fentanyl is typically used to treat patients with severe or chronic pain. In contrast, “illegal” fentanyl has frequently been added to other drugs including heroin, cocaine, methamphetamine, and MDMA to increase their potency and contributing to overdose deaths. In this study, we have investigated trends in fentanyl and fentanyl analogs positivity at a large academic medical center.

Methods
Retrospective analysis of results reported over seven years (2015–2021) for untargeted urine drug screens included testing performed for acute exposure and compliance. Presence of fentanyl, fentanyl analogs, and other prescribed or illicit used drugs was determined by gas chromatography—mass spectrometry (GCMS). Overall positivity was calculated as a percentage of total reported results.

Results
Fentanyl, and fentanyl analog positivity was determined for 40 782 urine drug screen results. Overall positivity for fentanyl and fentanyl analogs increased 3.8-fold over the past seven year from 1.53% (2015) to 5.78% (2021). Co-positivity with other illicit drugs such as cocaine and heroin also increased from 7% and 8% respectively, in 2015, to 42% and 13%, in 2021. Presence of fentanyl analogs such as fluorofentanyl, despropionylfentanyl, and acetylfentayl increased in the recent years as well.

Conclusion
Our data reveals increased positivity of fentanyl and fentanyl analogs present with cocaine, heroin, or their metabolites suggesting increased use of illicit fentanyl. The high prevalence supports the inclusion of fentanyl on routine urine drug screen immunoassays, or at a minimum as a reflex with a positive opiate screen.
B-303 Comparison of Tacrolimus Concentrations Obtained By the Newly Approved Alinity i Tacrolimus Assay With a Reference Liquid-Chromatography Combined With Tandem Mass spectrometry Assay
Authors: Woodard K , Kisler T , Dasgupta A 
Published: September 2023

Abstract

Background
Tacrolimus belongs to the calcineurin-inhibitor class of medications and used for preventing and maintenance of organ rejection in transplant recipients alone or in combination with other medications. Due to narrow therapeutic window and significant nephrotoxicity, this drug is routinely monitored with a suggested therapeutic range of 5–15 ng/mL. Recently, Abbott Laboratories (Abbott Park, IL) received FDA approval for using tacrolimus immunoassay on the Alinity i platform. We compared tacrolimus values in 101 transplant patients using Alinity i analyzer and a reference method based on liquid chromatography combined with mass tandem spectrometry (LC-MS/MS).

Methods
Tacrolimus immunoassay on the Alinity i analyzer is a chemiluminescent microparticle immunoassay (CMIA) requiring pretreatment of whole blood specimen prior to analysis. The analytical measurement range of the assay is 2–30 ng/mL. The left-over blood specimens were sent to Eurofins-Viracor reference laboratory in Lenexa, KS for analysis using LC-MS/MS method. After liquid-liquid extraction, specimens were analyzed using ascomycin as the internal standard for the presence of tacrolimus (assay is capable of analyzing other immunosuppressants simultaneously). UPLC C-18 column (waters) was used for separation using two mobile phases (A: 2.0 mM ammonium acetate in 0.1% formic caid in water and B: 2.0 mM ammonium acetate in 0.1% formic acid in methanol). Mass spectrometer was operated in positive ionization electrospray mode (tacrolimus m/z 821.6 &gt; 768.6, ascomycin m/z 809.756.6.

Results
The total precision of the low control was 3.6% (mean: 4.48 ng/mL; CV 0.16 ng/mL, n = 20) while the total precision of the high control was 5.1% (mean: 25.4 ng/mL, SD: 1.29 ng/mL, n = 20). The linearity of the assay (2.0–30.0 ng/mL) was verified by running various calibrators as unknown. Comparing tacrolimus values in specimens from 101 transplant patients (49 kidney, 19 bone marrow transplant, 16 liver and 15 heart transplant) using the reference method (LC-MS/MS) as the x-axis and the corresponding values obtained by the tacrolimus immunoassay using Alinity I analyzer, we observed the following regression equation: y = 0.97 x + 1.005 (n = 101, r = 0.95). For kidney transplant recipients, the regression equation was y = 0.96 + 1.20 (n = 49, r = 0.94).

Discussion
Excellent correlation between tacrolimus values obtained by the newly FDA approved tacrolimus immunoassay with the reference LC-MS/MS based assay indicates further improvement in the performance of immunoassays for therapeutic drug monitoring of immunosuppressants. This may be due to reduced cross-reactivity of metabolites as indicated in the package insert.

Conclusions
The newly approved tacrolimus immunoassay for the application on the Alinity I analyzer can be used for routine therapeutic drug monitoring of tacrolimus in clinical laboratories.
B-311 Increasing Xylazine Findings in Clinical Samples
Authors: Reinhart A 
Published: September 2023

Abstract

Background
Xylazine is a veterinary tranquilizer first appearing in illicit fentanyl preparations in 2019. It is believed to extend a fentanyl high and is now more commonly used as its own drug of abuse. Clinically, xylazine is of concern as it does not respond to naloxone or other antidote during overdoses. There are many forensic findings of increasing xylazine use since its original appearance, but little in the literature concerning clinical xylazine findings, possibly because xylazine is rarely screened in routine drug screening. We sought to find clinical cases with positive xylazine results from 2019–2022. Determined data included demographics (where possible to discern), sex distribution, age ranges, and frequency.

Methods
We queried our laboratory information management system (LIMS) for reported xylazine findings under expanded clinical drug screen utilizing liquid chromatography time of flight mass spectrometry (LC-TOF) for screening and liquid chromatography tandem mass spectrometry (LC-MS/MS) for confirmation, and specific xylazine testing utilizing LC-MS/MS.

Results
In 1928 clinical cases reported between January 2019 and January 2023, the xylazine positivity rate increased from 1.28% (2021; n = 13 positive xylazine results) to 5.05% positivity (2022; n = 29 positive xylazine results) despite a significant decrease in panels and tests with xylazine ordered (n = 1008 in 2021, n = 574 in 2022). The ages of the patients ranged 0–57 years; eleven cases concerned pediatric patients. Of cases that indicated the sex of the patient, 69% were male (n = 29). These cases were found in ten different states (AZ, CT, DE, IN, MA, NJ, OH, PA, UT, VA), with 18 incidental findings. While other drugs were present in incidental finding cases, all cases with an incidental positive xylazine result were also positive for fentanyl and norfentanyl, and several cases were also positive for fentanyl analogues, most commonly 4-ANPP, which occurred in 50% of these cases.

Conclusion
Xylazine use continues to be on the rise and should be of concern for emergency medicine doctors, poison control centers, and clinical laboratories.
B-314 Evaluation of Everolimus QMS Immunoassay on Roche Cobas c502 and Indiko Plus Analyzers
Authors: Buggs V , Song L , Metushi I 
Published: September 2023

Abstract

Background
Everolimus has a narrow therapeutic window and significant intra-individual variability. Rapid turn-around-time and precise results are critical for therapeutic drug monitoring and adequate dose adjustment. The Thermo Scientific Quantitiative Microsphere System (QMS®) everolimus immunoassay is the only FDA-approved immunoassay for measuring everolimus in human blood. The analytical performance of the Thermo Scientific QMS® everolimus immunoassay on Indiko Plus and Roche c502 platforms were evaluated and compared to LC-MS/MS.

Methods
Whole blood EDTA-anticoagulated patient samples were pretreated with methanol and precipitation reagent (immunoassay) or zinc sulfate/methanol (LC-MS/MS) to remove proteins before analysis. Imprecision was determined by running quality control and patient samples 20 times. Fifty transplant samples were used for method comparison. Deming regression, correlation coefficient (R2), and % bias plot analysis were performed using EP Evaluator® software.

Results
Imprecision (%CV) for QMS everolimus assay on Roche c502 was 16.7% (mean = 4.4 ng/mL) and 9.7% (mean = 10.3 ng/mL) for patient pools, and 16.5% (mean = 4.7 ng/mL), 10.3% (mean = 7.8 ng/mL), 10.7% (mean = 14.5 ng/mL) for quality control. QMS everolimus assay imprecision on Indiko Plus platform had CV of 3.1% (mean = 4.3 ng/mL), 3.3% (mean = 9.3 ng/mL), 2.8% (mean 14.3 ng/mL) for liver transplant patients and 3.3% (mean = 4.2 ng/mL), 3.3% (mean 8.3 ng/mL), 3.1 % (mean 15.6 ng/mL) for quality control. Patient comparison studies between Roche c502 vs LC-MS/MS revealed a R2 of 0.82, slope of 1.3 and %bias of −2.7%. QMS on Indiko Plus vs LC-MS/MS revealed a R2 of 0.84, slope of 0.96 and %bias of −21.9%.

Conclusions
The Indiko Plus platform had a constant negative bias vs LC-MS/MS while the Roche’s platform had lower average bias but it resulted in greater scatter in the therapeutic range (3–8 ng/mL) vs LC-MS/MS. Overall, we observed increased imprecision on the c502 analyzer.
B-334 How to Prevent the Next Wave in Overdose Crisis?—Detect Designer Benzodiazepines Using CEDIA and DRI Assays
Authors: Wu J , Ouyang A , Chua P , Ye L , Patel R 
Published: September 2023

Abstract

Background
Benzodiazepines are a class of drugs used for relieving symptoms of anxiety, insomnia, agitation, muscle spasms, and alcohol withdrawal. Benzodiazepines are CNS depressants enhancing the effect of GABA neurotransmitter at GABA receptors and they are well-known for drug abuse potential and drug dependence.Designer benzodiazepines are new psychoactive substances emerging in the past two decades. Designer benzodiazepines are often taken by individuals who use hallucinogenic and stimulant drugs. The drug abuse can lead to respiratory depression, coma, or death. There exists a need to develop a new homogeneous enzyme immunoassay. Existing commercial immunoassay antibodies may also cross-react with designer benzodiazepines which have chemical structure closely related to classic therapeutic benzodiazepines. Herein, we evaluated designer benzodiazepines cross-reactivity using DRI Benzodiazepines and CEDIA Benzodiazepines assays and demonstrated the potential of detecting designer benzodiazepines in urines.

Methods
DRI assay is based on competition between drug labeled with glucose-6-phosphate dehydrogenase (G6PDH) and free drug in the urine sample for a fixed amount of antibody binding sites. The enzyme activity is determined spectrophotometrically at 340 nm by measuring its ability to convert NAD to NADH. CEDIA technology is based on the bacterial enzyme β-galactosidase which has been genetically engineered into two inactive fragments, Enzyme Acceptor (EA) and Enzyme Donor (ED). The enzyme activity is then determined spectrophotometrically at 570 nm. Designer Benzodiazepines tested in this study are 3-Hydroxyphenazepam, Adinazolam, Bromazolam, Clonazolam, Cloniprazepam, Deschloroetizolam, Diclazepam, Etizolam, Flubromazepam, Flubromazolam, Flunitrazolam, Meclonazepam, N-Desmethylflunitrazepam, Nifoxipam, Nimetazepam, Nitrazolam, and Pyrazolam. Compounds were spiked into drug free urine and cross-reactivity was evaluated using DRI® Benzodiazepine Assay and CEDIA® Benzodiazepine Assays on the Beckman Coulter AU680.

Results
14 out of 17 (82%) tested designer benzodiazepines showed high cross-reactivity (&gt;100%) in DRI benzodiazepine assay. 12 out of 17 (71%) tested designer benzodiazepines showed high cross-reactivity (&gt;100%) in CEDIA assays with 200 ng/mL cutoff and 300 ng/mL cutoff. Compared to NPS-benzodiazepines trend data, this study demonstrated that DRI and CEDIA benzodiazepine assays can detect more than 50% (10 out of 19) designer benzodiazepines over 2020 Q1 to 2022 Q4 in the United States.

Conclusion
DRI and CEDIA Benzodiazepine Assays showed high cross reactivity of most seventeen designer benzodiazepines. The results provide potential tools for detect emerging designer benzodiazepines. Note: This was preliminary data that has not been reviewed by any regulatory authorities/cleared by FDA.
B-335 Rapid Quantitative Clozapine Measurements at the Point-of-Care Using Capillary Blood
Authors: Christenson R H , Hilaire M R , Salamone S J , Kelly D L 
Published: September 2023

Abstract

Background
Clozapine is the most effective medication for treatment-resistant schizophrenia, and is endorsed in professional guidelines. Therapeutic drug monitoring (TDM) is recommended for Clozapine due to high pharmacokinetic variability and risk of adverse drug reactions. The effectiveness of TDM for managing clozapine patients is limited by the lack of immediate results.

Methods
A novel immunoassay, the MyCare Insite Clozapine Test (MCI), was developed that uses 20 µL of capillary blood (CP) and delivers 7-minute quantitative results for a portable point-of-care (POC) device. To validate the MCI system, matching CP and serum samples were collected from 113 patients at two sites. CP was tested immediately with MCI and compared to serum immunoassay results on a Beckman-Coulter AU480 analyzer. Precision was evaluated using three whole blood spiked samples and two assay controls over five days with five replicates per day.

Results
The figure shows a correlation plot evaluated by Passing-Bablok regression of the POC MCI (y-axis) and the Beckman analyzer (x-axis). The regression line was y = 1.05*x + 1.4 ng/L, r = 0.927, indicating good agreement. Precision studies demonstrated that repeatability was ≤11% and within-lab precision was ≤13%.

Conclusion
Quantitative Clozapine results are available in 7 min using CP in a portable POC device. A rapid POC alternative can achieve timely patient results in many testing environments. Rapid Clozapine testing has the potential for expanding utilization of this important drug by providing actionable POC results.
B-339 A Clinical Research LC-MS/MS Method for the Analysis of Antidepressants in Plasma
Authors: Balloch S , Calton L , Hammond G 
Published: September 2023

Abstract

Background
Depression is common globally, with an estimated 3.8% of the population affected. The condition can impact individuals’ ability to function in work, social and family settings. Many antidepressant drugs are currently prescribed, encompassing selective serotonin reuptake inhibitors (SSRIs), serotonin-nonresponsive reuptake inhibitors (SNRIs) and tetracyclic antidepressants (TeCAs). However pharmacokinetic and drug interactions are known, therefore a reliable quantitative clinical research method may play a role in researching the effects of their administration. Waters has developed a clinical research method for the following antidepressants in plasma; citalopram, desmethylfluoxetine, duloxetine, fluoxetine, fluvoxamine, O-desmethylvenlafaxine, sertraline and venlafaxine (10–1000 ng/mL); mirtazapine (5–500 ng/mL) and trazadone (30–3000 ng/mL).

Methods
Matrix matched calibrators and QCs were prepared using in-house stocks and pooled plasma. Samples (50 µL) were treated with internal standard in acetonitrile. A water/methanol/ammonium acetate gradient was used with a Waters™ XSelect™ Premier HSS T3 column on a Waters ACQUITY™ UPLC™ I-Class followed by detection on a Xevo® TQD mass spectrometer in a 5 min run.

Results
No system carryover was observed following analysis of plasma samples containing the highest concentration calibrators. Analytical sensitivity investigations indicated precise quantification (≤20% CV, ≤17.6% bias) at concentrations equal to or lower than the lowest concentration calibrator. Total precision and repeatability were assessed (3 pools, 5 replicates, 5 days; n = 25) and determined to be ≤10.0% RSD. Linearity experiments determined the method provided first or second order fits over the ranges analyzed; additionally, each run met acceptance criteria (coefficient of correlation ≥ 0.995, determined concentrations of calibrators ±15% of nominal, ±20% in the case of the lowest calibrator). Post-column infusion experiments demonstrated analytes eluted in regions free of major ion suppression or enhancement. Evaluation of matrix effects at low and high concentrations indicated compensation by the internal standard. Addition of high concentrations of several endogenous and exogenous materials did not affect quantification.

Conclusion
This quantitative method for clinical research demonstrates very good precision with minimal matrix effects and allows for the multiplexing of a panel of antidepressants in plasma in a short run time. For Research Use Only. Not for Use in Diagnostic Procedures.
B-340 Performance of a Quality System for Targeted Monoclonal Antibodies in a Reference Clinical Pharmacokinetic Laboratory
Authors: McFarland J , McLachlan O , Tsang J , Gowhari M , Ho J , Hughes P , Dervieux T 
Published: September 2023

Abstract

Background
The homogenous mobility shift assay (HMSA) laboratory developed test (LDT) quantifies monoclonal antibody drug and anti-drug antibody levels and is useful for the management of inflammatory bowel diseases (IBD). We report the implementation and performance of a quality system for these LDTs, as well as longitudinal trends of exposure and immunization prevalence from a reference clinical pharmacokinetic (PK) laboratory (Prometheus Laboratories, Inc.).

Methods
Quality control (QC) HMSA data was evaluated to verify conformance to specifications for infliximab (IFX), adalimumab (ADA), vedolizumab (VDZ), ustekinumab (UST) and their respective anti-drug antibodies (antibody to IFX [ATI], ADA [ATA], VDZ [ATV], and UST [ATU]). A QC panel consisting of five serum specimens spanning the reportable range per each drug or anti-drug antibody LDT was established, stored at subzero temperature, and tested monthly over a period of two years. Acceptable performance consisted of 80%–120% accuracy from the target value. All specimens were blinded to operator and processed with the clinical PK load of testing. Exposure and immunization prevalence over three years of testing was evaluated. All patient data were de-identified before analysis. Median drug level (interquartile ranges [IQR]), and anti-drug percent positivity by international classification of disease was also estimated (ICD-10 based).

Results
The QC results establish that the drug (IFX, ADA, VDZ, UST) and anti-drug (ATI, ATA, ATV, ATU) LDTs report acceptable performance with 80%–120% accuracy over the two-year evaluation period. Median drug concentrations with IQR were assessed for the clinical sample set differentiated by disease classification as depicted in Fig. 1. Anti-drug positivity levels were higher for ATI (mean: 11.5%) and ATA (mean: 13.6%) than those from ATV (mean 2.0%) and ATU (mean: 0.6%).

Conclusion
The QC data presented supports the performance of HMSA for therapeutic drug monitoring (TDM) of IBD and other immune mediated inflammatory diseases.