Screening Panel Research Articles

Smart Arduino Touch Switch Board for Home Automation

We live in an exciting time where more and more everyday items “things” are becoming smart! “Things” have sensors and can communicate with other “things” and can provide control to more “things”. The Internet of Things, IoT, is totally up to us people in a huge way and we are rapidly inventing new gadgets to enhance our lives. The prices of microcontrollers used in IoT devices with the ability to talk over a network keeps reducing and developers can now think and build things inexpensively. This IoT based home automation project is done using low-cost custom build Arduino UNO module, and Bluetooth module. A system that uses mobile app to control basic home functions and features automatically through mobile from anywhere in the house, an automated home is sometimes called a smart home. The home automation system differs from other IoT based systems by allowing a person or the user to operate the system from anywhere in the house through wireless connection without breaking any convention usability. This project provides significant electric power solution at homes and at the offices with help of dynamic control of electric equipments over the wireless network and using smart switch, and another is a low-cost touch screen panel. Project contains both hardware and software development where hardware program is built on Arduino IDE, whereas mobile application is built using MIT App Development. Key Word: Home automation, Bluetooth, Touch switch.

Comparision of the diagnostic sensitivity of biochemical tests in the screening of monoclonal gammopathy

: Monoclonal gammopathy include several clinical variants ranging from asymptomatic MGUS, asymptomatic smouldering myeloma, multiple myeloma to aggressive plasma cell leukemia. The characteristic property of myeloma cells is the production and secretion of M protein. Due to its diverse structure, no single test can identify M protein accurately in all patients. A simple, non-invasive combination of tests is necessary for the screening of monoclonal gammopathy. The present study aims to assess the diagnostic sensitivity of the biochemical tests used in the screening of monoclonal gammopathy.: This study was conducted retrospectively of the newly diagnosed cases of monoclonal gammopathy screened with SPE and ISUB/IT using sebia capillary electrophoresis and serum free light chain ratio using immunoturbidimetry method.: Out of the 142 patients included in the study, 120 had M band in SPE, 125 had monoclonal gammopathy by ISUB/IT and 121 patients had abnormal sFLCR. The diagnostic sensitivity obtained was 84.5% 88% and 85.2% for SPE, ISUB/IT and sFLCR respectively. Addition of sFLCR to SPE and ISUB/IT could identify 20 patients who had no abnormality in either of these tests. The combined sensitivity of SPE and ISUB/IT was 88%, while that of SPE and sFLCR and a combination of all the three tests was 98.6%. The simple, non-invasive, cost-effective screening panel of SPE plus sFLC ratio could be used as the initial screening method for patients with suspected monoclonal gammopathy with increased reliability.

Prospective Observational Study of Prevalence, Assessment and Treatment of Pancreatic Exocrine Insufficiency in Patients with Inoperable Pancreatic Malignancy (PANcreatic Cancer Dietary Assessment-PanDA).

Pancreatic exocrine insufficiency (PEI) in patients with advanced pancreatic cancer (aPC) is well documented, but there is no consensus regarding optimal screening. Patients diagnosed with aPC referred for palliative therapy were prospectively recruited. A full dietetic assessment (including Mid-Upper Arm Circumference (MUAC), handgrip and stair-climb test), nutritional blood panel, faecal elastase (FE-1) and 13C-mixed triglyceride breath tests were performed. prevalence of dietitian-assessed PEI (demographic cohort (De-ch)); design (diagnostic cohort (Di-ch)) and validation (follow-up cohort (Fol-ch)) of a PEI screening tool. Logistic and Cox regressions were used for statistical analysis. Between 1 July 2018 and 30 October 2020, 112 patients were recruited (50 (De-ch), 25 (Di-ch) and 37 (Fol-ch)). Prevalence of PEI (De-ch) was 64.0% (flatus (84.0%), weight loss (84.0%), abdominal discomfort (50.0%) and steatorrhea (48.0%)). The derived PEI screening panel (Di-ch) included FE-1 (normal/missing (0 points); low (1 point)) and MUAC (normal/missing (>percentile 25) (0 points); low (2 points)) and identified patients at high-risk (2-3 total points) of PEI [vs. low-medium risk (0-1 total points)]. When patients from the De-ch and Di-ch were analysed together, those classified by the screening panel as "high-risk" had shorter overall survival (multivariable Hazard Ratio (mHR) 1.86 (95% CI 1.03-3.36); p-value 0.040). The screening panel was tested in the Fol-ch; 78.4% patients classified as "high-risk", of whom 89.6% had dietitian-confirmed PEI. The panel was feasible for use in clinical practice (64.8% patients completed all assessments), with high acceptability (87.5% would repeat it). Most patients (91.3%) recommended dietetic input for all patients with aPC. PEI is present in most patients with aPC; early dietetic input provides a holistic nutritional overview, including, but not limited to, PEI. This proposed screening panel may help to prioritise those at higher risk of PEI, requiring urgent dietitian input. Its prognostic role needs further validation.

Infants with Congenital Diseases Identified through Newborn Screening—United States, 2018–2020

Newborn screening (NBS) is a state or territory-based public health system that screens newborns for congenital diseases that typically do not present with clinical symptoms at birth but can cause significant mortality and morbidity if not detected or treated quickly. NBS continues to be one of the most successful public health interventions in the US, providing early detection and intervention to all infants. The increase in overall birth prevalence of core Recommended Uniform Screening Panel (RUSP) diseases detected via dried blood spot (DBS) specimens from 2015-2017 (17.50-18.31 per 10,000) to 2018-2020 (20.07 per 10,000), as reported into the APHL NewSTEPs database, affirms the importance and impact of NBS programs. This report presents aggregate numbers and birth prevalence of diseases detected by DBS on the RUSP from 2018-2020, including data from fifty US states and two territories.

Utility of anti-immunoglobulin IgA, IgG, IgM, Kappa, Lambda - FITC (conjugate) cocktail in routine renal pathology practice

BackgroundImmunofluorescence studies on frozen sections are an essential component in the evaluation of renal biopsies. The basic panel in most centres constitutes IgG, IgA, IgM, C3c, C1q, Kappa and Lambda light chain antibody testing. The purpose of this panel is to detect immunoglobulin or complement deposits and further subclassify the disease based on the location, intensity and pattern of immunoglobulin and complement staining. However, there are a substantial proportion of nephropathies that do not show any obvious immune-deposits on immunofluorescence. We currently, do not have any evidence-based alternative immunofluorescence panel to rule out these conditions. This study aims to evaluate the utility of anti-immunoglobulin IgA, IgG, IgM, Kappa, Lambda - FITC cocktail immunofluorescence on renal biopsy frozen sections with emphasis on its role as a primary screening panel in conjunction with C3c and C1q.MethodsAnti-immunoglobulin IgA, IgG, IgM, Kappa, Lambda light chain - FITC cocktail immunofluorescence was performed on 593 consecutive native renal biopsies along with the routine panel comprising of the individual FITC labelled IgG, IgA, IgM, C3c, C1q, Kappa and Lambda light chain immunofluorescence stains.ResultsIn 235 (39.6%) cases immune deposits (immune-complex mediated and monoclonal gammopathy-related) were present and the rest 354 (59.7%) cases were negative for immunoglobulin or complement deposits. Overall, the sensitivity, specificity, positive predictive value and negative predictive values of anti-immunoglobulin IgA, IgG, IgM, Kappa and Lambda - FITC cocktail in distinguishing immune-complex/immunoglobulin-mediated glomerulopathies from non-immune complex/immunoglobulin-mediated glomerulopathies were 100% each.ConclusionAnti-immunoglobulin IgA, IgG, IgM, Kappa and Lambda - FITC cocktail when used in conjunction with C3c and C1q, can be an effective first line investigation in all native renal biopsies. Further, testing with the individual FITC labelled IgG, IgA, IgM, Kappa and Lambda light chain immunofluorescence can be performed, depending on the initial screening as described above. Overall, this algorithmic approach can save valuable resources.

Self Capacitance Mismatch Calibration Technique for Fully-Differential Touch Screen Panel Self Capacitance Sensing System

This paper presents a fully-differential touch screen panel (TSP) self-capacitance sensing (SCS) system with a self-capacitance mismatch calibration technique. Due to the self-capacitance mismatch of TSP, the analog front-end (AFE) of the receiver (RX) circuit suffers from dynamic range degradation and gain limitations, which lead to the signal-to-noise ratio (SNR) loss for the TSP SCS system. The proposed calibration introduces the difference in input resistance and the driving amplifier's strength between the fully-differential input. Thus, the mismatch effect is efficiently relieved in terms of area and power consumption. The proposed calibration restores the SNR by 19.54 dB even under the worst self-capacitance mismatch case.

Implementation of Newborn Screening for Conditions in the United States First Recommended during 2010-2018.

The Recommended Uniform Screening Panel (RUSP) is the list of conditions recommended by the US Secretary of Health and Human Services for inclusion in state newborn screening (NBS). During 2010-2022, seven conditions were added to the RUSP: severe combined immunodeficiency (SCID) (2010), critical congenital heart disease (CCHD) (2011), glycogen storage disease, type II (Pompe) (2015), mucopolysaccharidosis, type I (MPS I) (2016), X-linked adrenoleukodystrophy (X-ALD) (2016), spinal muscular atrophy (SMA) (2018), and mucopolysaccharidosis, type II (MPS II) (2022). The adoption of SCID and CCHD newborn screening by programs in all 50 states and three territories (Washington, D.C.; Guam; and Puerto Rico) took 8.6 and 6.8 years, respectively. As of December 2022, 37 programs screen for Pompe, 34 for MPS I, 32 for X-ALD, and 48 for SMA. The pace of implementation based on the average additional number of NBS programs per year was most rapid for SMA (11.3), followed by CCHD (7.8), SCID (6.2), MPS I (5.4), Pompe (4.9), and X-ALD (4.7).

Abstract 198: Pan-cancer assay-ready organoid drug screening with robust, reproducible and clinically-relevant output

Abstract Introduction The pharmaceutical industry has mostly relied on 2D cancer cell lines and 3D spheroids for in vitro testing, but poor correlation of pre-clinical and clinical outcomes has driven the development of more predictive models. Patient-derived organoids (PDOs) have emerged as representative in vitro avatars of tumor biology, allowing the generation of biobanks covering a large variety in indications and genetic backgrounds. Here, we assess the robustness of our drug screening platform by testing the reproducibility of our organoid assays within and between organoid batches, read-outs, and different labs. We present an assay-ready organoid platform, allowing short timelines, repeated assays from a single batch of organoids, high throughput assays and large panel screens. Methods Colorectal, breast, lung, pancreatic, ovarian, cervix and melanoma PDO models were selected for subtype and driver mutation variety, banked in large batches, and preserved in assay-ready format. Outgrowth and drug responses of these batches were compared to those of organoids produced in the classic method. Experiments were executed using automated liquid handling equipment to standardize procedures and increase consistency. Performance of the drug screens were assessed by repeated drug sensitivity testing and by calculating intra- and interplate variability, control variability (CV), Z-factor, assay windows and IC50 values. Moreover, drug responses were tested in both celltiterglo-based and high content imaging-based assays. Results Assay performance of the organoid drug testing platform was high, with high Z-factors (>0.6), and low intra- and interplate variability (CV <15%) indicating reproducible assays. Both the classic and assay-ready organoid technologies resulted in highly reproducible IC50outputs, also when assays were executed months apart or in different labs. Timelines were shortened from months to weeks, and the reduced logistical burden allows screening of large organoid panels of >50 models. Drug sensitivity testing in our fast-paced panel screening platform distinguished sensitive from partially and insensitive models, illustrating how organoids allow patient stratification. Conclusion and Discussion The assay-ready organoid technology presented here has further improved our already highly predictive and reproducible organoid drug testing platform. The resulting short timelines and large panel screening capabilities will further unlock the great potential of PDO technology. Using large organoid panels in pre-clinical drug development and in combination with biomarker analysis will allow identification of responsive indications, subtypes, and genotypes, and for early patient stratification. Moreover, as most models presented in our panel screen platform are also available as PDX models, it allows for easy translation into in vivo follow-up studies. Citation Format: Liza Wijler, Jara Garcia Mateos, My Nguyen, Annelot Staes, Linda van Seters, Lama Alhaj Hasan, Victor Tiroille, Bram Herpers, Leo Price, Mariusz Madej, Michiel Fokkelman, Marrit Putker. Pan-cancer assay-ready organoid drug screening with robust, reproducible and clinically-relevant output [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 198.

Abstract 231: Evaluation of a 293 gene oncology-focused exome sequencing panel for screening of ovarian cancer patients for BRCA mutations as a prerequisite for autologous cellular immunotherapy manufacturing

Abstract Gemogenovatucel-T is a cellular immunotherapy manufactured from harvested tumor tissue transfected with a plasmid which encodes granulocyte macrophage colony stimulating factor (GM-CSF) and a short harpin RNA which specifically reduces the expression of furin and downstream targets TGF-β1, and TGF- β2. In the Phase IIB VITAL study involving stage 3/4 ovarian cancer patients, recurrence-free survival and overall survival in patients who were BRCA1/2 wild-type was improved in patients receiving Gemogenovatucel-T as compared to placebo. The same improvements in recurrence-free and overall survival were not observed in patients with pathogenic or likely pathogenic mutations in BRCA1/2 genes. Similar improvements were demonstrated in patients who were homologous recombination proficient. For future clinical trials it is necessary to rapidly examine genomic DNA from blood of ovarian cancer patients to determine BRCA mutation status for eligibility to manufacture autologous Gemogenovatucel-T. An oligonucleotide hybridization-based capture panel targeting the coding exons and adjacent intronic regions of 293 oncology-related genes was developed for use in enriching DNA libraries molecules for sequencing on the Illumina platform. The purpose of the current study was to evaluate the utility of this panel for detection of likely pathogenic and pathogenic mutations in the BRCA1/2 genes. Genomic DNA from Gemogenovatucel-T preparations from the Phase IIB trial including 10 patients harboring known mutations in the BRCA1/2 genes and 2 BRCA wild type patients was processed through library preparation, hybridization-based capture using the 293 gene probe set, and sequencing on the Illumina NextSeq 2000. All eleven previously identified pathogenic mutations in BRCA1/BRCA2 gene, including 8 short insertions/deletions (INDELs) and 3 single nucleotide polymorphisms (SNPs) were detected with the method. No false positive pathogenic BRCA mutations were detected in any of the 12 samples. The panel was evaluated further using a set of reference genomic DNAs from Horizon Discovery containing known polymorphisms in BRCA as well as 30 other oncology-related genes. 100% of the expected SNPs and INDELs with an allele frequency of >3% were detected; no false positive likely pathogenic or pathogenic polymorphisms with an allele frequency of >2.5% were detected in the genomic DNA of healthy control patients. The 293 gene panel will be suitable for identification of BRCA mutations in patients to screen patients eligible for Gemogenovatucel-T manufacturing. The entire process starting with blood DNA extraction and ending with report generation can be completed in about 3.5 days. The results show that a comprehensive oncology-focused sequencing panel can be an effective tool for focused gene-specific variant analysis. Citation Format: David Willoughby, Ernie Bognar, Weiming Shen, Laura Nejedlik, Casey Nagel, Aman Pruthi, Peter Zhang, Adam Walter, Heidi Zupanc, Chris Jay, John Neumunaitis. Evaluation of a 293 gene oncology-focused exome sequencing panel for screening of ovarian cancer patients for BRCA mutations as a prerequisite for autologous cellular immunotherapy manufacturing [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 231.

Community data-driven approach to identify pathogenic founder variants for pan-ethnic carrier screening panels

BackgroundThe American College of Medical Genetics and Genomics (ACMG) recently published new tier-based carrier screening recommendations. While many pan-ethnic genetic disorders are well established, some genes carry pathogenic founder variants (PFVs) that are unique to specific ethnic groups. We aimed to demonstrate a community data-driven approach to creating a pan-ethnic carrier screening panel that meets the ACMG recommendations.MethodsExome sequencing data from 3061 Israeli individuals were analyzed. Machine learning determined ancestries. Frequencies of candidate pathogenic/likely pathogenic (P/LP) variants based on ClinVar and Franklin were calculated for each subpopulation based on the Franklin community platform and compared with existing screening panels. Candidate PFVs were manually curated through community members and the literature.ResultsThe samples were automatically assigned to 13 ancestries. The largest number of samples was classified as Ashkenazi Jewish (n = 1011), followed by Muslim Arabs (n = 613). We detected one tier-2 and seven tier-3 variants that were not included in existing carrier screening panels for Ashkenazi Jewish or Muslim Arab ancestries. Five of these P/LP variants were supported by evidence from the Franklin community. Twenty additional variants were detected that are potentially pathogenic tier-2 or tier-3.ConclusionsThe community data-driven and sharing approaches facilitate generating inclusive and equitable ethnically based carrier screening panels. This approach identified new PFVs missing from currently available panels and highlighted variants that may require reclassification.

Overexpression and Role of HHLA2, a Novel Immune Checkpoint, in Colorectal Cancer.

The study aimed to investigate correlations between HHLA2 levels and parameters, including microsatellite instability (MSI) status, CD8+ cells, and histopathological features: budding, tumor-infiltrating lymphocytes (TILs), TNM scale, grading, cytokines, chemokines, and cell signaling moleculesin colorectal cancer (CRC). Furthermore, the immune infiltration landscape and HHLA2-related pathways in colorectal cancer using available online datasets were analyzed. The study included 167 patients diagnosed with CRC. Expression of HHLA2 was detected by immunohistochemistry method (IHC) and enzyme-linked immunosorbent assay (ELISA). The IHC was used to evaluate the MSI and CD8+ status. The budding and TILs were measured using a light microscope. The concentrations of cytokines, chemokines, and cell signaling molecules were measured to analyze the data by the Bio-Plex Pro Human cytokine screening panel, 48 cytokine assay, and principal component analysis (PCA). Geneset enrichment analysis (GSEA) was conducted to identify HHLA2-related pathways. The biological function of HHLA2 was predicted by Gene Ontology (GO). Analysis of the immune infiltration landscape of HHLA2 in colorectal cancer was made by the web-based tool Camoip. High HHLA2 expression was detected in CRC tumor tissues compared to the adjacent noncancerous tissues. The percentage of HHLA2-positive tumors was 97%. GSEA and GO showed that HHLA2 upregulation correlated with cancer-related pathways and several biological functions. Tumor-infiltrating lymphocytes score correlated positively with IHC HHLA2 expression level percentage. There was a negative correlation between HHLA2, anti-tumor cytokines and pro-tumor growth factors. This study provides a valuable insight into the role of HHLA2 in CRC. We reveal the role of HHLA2 expression as well as a stimulatory and inhibitory immune checkpoint in colorectal cancer. Further research may verify the therapeutic values of the HHLA2-KIR3DL3/TMIGD2 pathway in colorectal cancer.

Human Polo-like Kinase Inhibitors as Antiplasmodials.

Protein kinases have proven to be a very productive class of therapeutic targets, and over 90 inhibitors are currently in clinical use primarily for the treatment of cancer. Repurposing these inhibitors as antimalarials could provide an accelerated path to drug development. In this study, we identified BI-2536, a known potent human polo-like kinase 1 inhibitor, with low nanomolar antiplasmodial activity. Screening of additional PLK1 inhibitors revealed further antiplasmodial candidates despite the lack of an obvious orthologue of PLKs in Plasmodium. A subset of these inhibitors was profiled for their in vitro killing profile, and commonalities between the killing rate and inhibition of nuclear replication were noted. A kinase panel screen identified PfNEK3 as a shared target of these PLK1 inhibitors; however, phosphoproteome analysis confirmed distinct signaling pathways were disrupted by two structurally distinct inhibitors, suggesting PfNEK3 may not be the sole target. Genomic analysis of BI-2536-resistant parasites revealed mutations in genes associated with the starvation-induced stress response, suggesting BI-2536 may also inhibit an aminoacyl-tRNA synthetase.

SARS-CoV-2 spike protein induces IL-18-mediated cardiopulmonary inflammation via reduced mitophagy

Cardiopulmonary complications are major drivers of mortality caused by the SARS-CoV-2 virus. Interleukin-18, an inflammasome-induced cytokine, has emerged as a novel mediator of cardiopulmonary pathologies but its regulation via SARS-CoV-2 signaling remains unknown. Based on a screening panel, IL-18 was identified amongst 19 cytokines to stratify mortality and hospitalization burden in patients hospitalized with COVID-19. Supporting clinical data, administration of SARS-CoV-2 Spike 1 (S1) glycoprotein or receptor-binding domain (RBD) proteins into human angiotensin-converting enzyme 2 (hACE2) transgenic mice induced cardiac fibrosis and dysfunction associated with higher NF-κB phosphorylation (pNF-κB) and cardiopulmonary-derived IL-18 and NLRP3 expression. IL-18 inhibition via IL-18BP resulted in decreased cardiac pNF-κB and improved cardiac fibrosis and dysfunction in S1- or RBD-exposed hACE2 mice. Through in vivo and in vitro work, both S1 and RBD proteins induced NLRP3 inflammasome and IL-18 expression by inhibiting mitophagy and increasing mitochondrial reactive oxygenation species. Enhancing mitophagy prevented Spike protein-mediated IL-18 expression. Moreover, IL-18 inhibition reduced Spike protein-mediated pNF-κB and EC permeability. Overall, the link between reduced mitophagy and inflammasome activation represents a novel mechanism during COVID-19 pathogenesis and suggests IL-18 and mitophagy as potential therapeutic targets.

Testing Road Vehicle User Interfaces Concerning the Driver’s Cognitive Load

This paper investigates the usability of touch screens used in mass production road vehicles. Our goal is to provide a detailed comparison of conventional physical buttons and capacitive touch screens taking the human factor into account. The pilot test focuses on a specific Non-driving Related Task (NDRT): the control of the on-board climate system using a touch screen panel versus rotating knobs and push buttons. Psychological parameters, functionality, usability and, the ergonomics of In-Vehicle Information Systems (IVIS) were evaluated using a specific questionnaire, a system usability scale (SUS), workload assessment (NASA-TLX), and a physiological sensor system. The measurements are based on a wearable eye-tracker that provides fixation points of the driver’s gaze in order to detect distraction. The closed road used for the naturalistic driving study was provided by the ZalaZONE Test Track, Zalaegerszeg, Hungary. Objective and subjective results of the pilot study indicate that the control of touch screen panels causes higher visual, manual, and cognitive distraction than the use of physical buttons. The statistical analysis demonstrated that conventional techniques need to be complemented in order to better represent human behavior differences.

Determining the most accurate 16S rRNA hypervariable region for taxonomic identification from respiratory samples

16S rRNA gene profiling, which contains nine hypervariable regions (V1–V9), is the gold standard for identifying taxonomic units by high-throughput sequencing. Microbiome studies combine two or more region sequences (usually V3–V4) to increase the resolving power for identifying bacterial taxa. We compare the resolving powers of V1–V2, V3–V4, V5–V7, and V7–V9 to improve microbiome analyses in sputum samples from patients with chronic respiratory diseases. DNA were isolated from 33 human sputum samples, and libraries were created using a QIASeq screening panel intended for Illumina platforms (16S/ITS; Qiagen Hilden, Germany). The analysis included a mock community as a microbial standard control (ZymoBIOMICS). We used the Deblur algorithm to identify bacterial amplicon sequence variants (ASVs) at the genus level. Alpha diversity was significantly higher for V1–V2, V3–V4, and V5–V7 compared with V7–V9, and significant compositional dissimilarities in the V1–V2 and V7–V9 analyses versus the V3–V4 and V5–V7 analyses. A cladogram confirmed these compositional differences, with the latter two being very similar in composition. The combined hypervariable regions showed significant differences when discriminating between the relative abundances of bacterial genera. The area under the curve revealed that V1–V2 had the highest resolving power for accurately identifying respiratory bacterial taxa from sputum samples. Our study confirms that 16S rRNA hypervariable regions provide significant differences for taxonomic identification in sputum. Comparing the taxa of microbial community standard control with the taxa samples, V1–V2 combination exhibits the most sensitivity and specificity. Thus, while third generation full-length 16S rRNA sequencing platforms become more available, the V1–V2 hypervariable regions can be used for taxonomic identification in sputum.

A non-linear model of screen panel for dynamics analysis of a flip-flow vibrating screen

A non-linear model of screen panel for dynamics analysis of a flip-flow vibrating screen

A multicenter prospective study of next-generation sequencing-based newborn screening for monogenic genetic diseases in China

BackgroundNewborn screening (NBS) is an important and successful public health program that helps improve the long-term clinical outcomes of newborns by providing early diagnosis and treatment of certain inborn diseases. The development of next-generation sequencing (NGS) technology provides new opportunities to expand current newborn screening methodologies.MethodsWe designed a a newborn genetic screening (NBGS) panel targeting 135 genes associated with 75 inborn disorders by multiplex PCR combined with NGS. With this panel, a large-scale, multicenter, prospective multidisease analysis was conducted on dried blood spot (DBS) profiles from 21,442 neonates nationwide.ResultsWe presented the positive detection rate and carrier frequency of diseases and related variants in different regions; and 168 (0.78%) positive cases were detected. Glucose-6-Phosphate Dehydrogenase deficiency (G6PDD) and phenylketonuria (PKU) had higher prevalence rates, which were significantly different in different regions. The positive detection of G6PD variants was quite common in south China, whereas PAH variants were most commonly identified in north China. In addition, NBGS identified 3 cases with DUOX2 variants and one with SLC25A13 variants, which were normal in conventional NBS, but were confirmed later as abnormal in repeated biochemical testing after recall. Eighty percent of high-frequency gene carriers and 60% of high-frequency variant carriers had obvious regional differences. On the premise that there was no significant difference in birth weight and gestational age, the biochemical indicators of SLC22A5 c.1400C > G and ACADSB c.1165A > G carriers were significantly different from those of non-carriers.ConclusionsWe demonstrated that NBGS is an effective strategy to identify neonates affected with treatable diseases as a supplement to current NBS methods. Our data also showed that the prevalence of diseases has significant regional characteristics, which provides a theoretical basis for screening diseases in different regions.

Validation of a targeted metabolomics panel for improved second-tier newborn screening.

Improved second-tier assays are needed to reduce the number of false positives in newborn screening (NBS) for inherited metabolic disorders including those on the Recommended Uniform Screening Panel (RUSP). We developed an expanded metabolite panel for second-tier testing of dried blood spot (DBS) samples from screen-positive cases reported by the California NBS program, consisting of true- and false-positives from four disorders: glutaric acidemia type I (GA1), methylmalonic acidemia (MMA), ornithine transcarbamylase deficiency (OTCD), and very long-chain acyl-CoA dehydrogenase deficiency (VLCADD). This panel was assembled from known disease markers and new features discovered by untargeted metabolomics and applied to second-tier analysis of single DBS punches using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in a 3-min run. Additionally, we trained a Random Forest (RF) machine learning classifier to improve separation of true- and false positive cases. Targeted metabolomic analysis of 121 analytes from DBS extracts in combination with RF classification at a sensitivity of 100% reduced false positives for GA1 by 83%, MMA by 84%, OTCD by 100%, and VLCADD by 51%. This performance was driven by a combination of known disease markers (3-hydroxyglutaric acid, methylmalonic acid, citrulline, and C14:1), other amino acids and acylcarnitines, and novel metabolites identified to be isobaric to several long-chain acylcarnitine and hydroxy-acylcarnitine species. These findings establish the effectiveness of this second-tier test to improve screening for these four conditions and demonstrate the utility of supervised machine learning in reducing false-positives for conditions lacking clearly discriminating markers, with future studies aimed at optimizing and expanding the panel to additional disease targets.

A rare case of clinically significant naturally occurring anti-Cw reported in a healthy male donor.

Anti-Cw antibody is an immunoglobulin against the red cell antigen Cw which is a low frequency red cell antigen that belongs to the Rh antigen system. It is a clinically significant antibody and may cause haemolysis on exposure to antigen positive red cells. Due to its low frequency, it is not included in routine antibody screening panels. A 32years healthy male donor with no history of transfusion donated whole blood at the department of Transfusion Medicine & Blood Centre of our institute. As a part of routine pre-transfusion testing, the donor's samples were subjected to automated blood grouping and screening for unexpected red cell antibodies using 3 cells panel on solid-phase red-cell adherence (SPRCA) (Galileo Neo, Immucor, Norcross, USA). The antibody screening came out to be positive with a reaction in cell I of the antibody screening panel. Further the antibody was identified as anti Cw in using 16 cells panel, select cells and phenotyping. In the present case, the anti-Cw antibody was found to be reactive at 37°C and AHG phase which could lead to haemolytic transfusion reaction. The fact that the male donor had no history of transfusion or transplant led us to the conclusion that it was a naturally occurring, but a clinically significant antibody. This case highlights the importance of performing an antibody screening for healthy donors as well and urges transfusion services to procure screening cells which incorporate Cw positive cells.

Ultrashort laser sintering of printed silver nanoparticles on thin, flexible, and porous substrates

The fabrication of low-cost and mechanically robust flexible electronic patterns has increasingly gained attention due to their growing applications in flexible displays, touch screen panels, medical devices, and solar cells. Such applications require cost-effective deposition of metals in a well-controlled manner potentially using nanoparticles (NPs). The presence of solvent and precursors in NP based inks impacts the electrical conductivity of the printed pattern and a post-processing heating step is typically performed to restore the electrical properties and structure of the material. We report printing with picolitre droplet volumes of silver (Ag) NPs on flexible substrates using an acoustic microdroplet dispenser. The low-cost, controlled deposition of Ag ink is performed at room temperature on photopaper, polyimide and clear polyimide substrates. A localized, ultrashort pulsed laser with minimal heat affected zone is employed to sinter printed Ag patterns. For comparison, oven sintering is performed, and the results are analysed with scanning electron microscopy, four-point probe and Hall measurements. The femtosecond laser sintering revealed highly organized, connected nanostructure that is not achievable with oven heating. A significant decrease in sheet resistance, up to 93% in Ag NPs on clear polyimide confirms the laser sintering improves the connectivity of the printed film and as a result, the electrical properties are enhanced. The surface morphology attained by the laser sintering process is interpreted to be due to a joining of NPs as a result of a solid-state diffusion process in the near surface region of NPs.