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An investigation into the distribution of radial immunodiffusion quality control data.

Quality control data from routine radial immunodiffusion assays for IgG, IgA, IgM, C3, C4 and alpha1-antitrypsin were tested by the Kolmogorov--Smirnov procedure for gaussian distribution. All but alpha1-antitrypsin were nongaussian in type. Further analysis of these date by plotting on log-normal probability paper showed them to have a log-normal distribution. Treatment of the date by either gaussian or nonparametric statistical methods produced little difference in confidence limits. It does not appear necessary to use nonparametric methods to calculate confidence limits from quality control data for the procedures studied.

Complement C3

Suitability of commercial control sera for the quality control of activity determination of alkaline phosphatase.

The suitability of thirteen commercially available control sera for measuring alkaline phosphatase (EC 3.1.3.1; orthophosphoric acid monoester phosphohydrolase, ALP) activity in human serum was tested. Apart from differences in ALP activity observed in some reconstituted commercial sera, the behaviour of control materials towards experimental variables such as the nature and concentration of the substrate, pH and type of buffer (or PO4-acceptor) together with the composition of the isoenzymes present in human serum highlights the problems and difficulties if commercial materials are to be used as control sera. The half-saturation constants in control sera were in all cases smaller than those of ALP isoenzymes from bone and liver. The shape of substrate activity curves and the pH optimum in most of control sera differed from that of human serum. The discrepant kinetic data of control materials and human serum may mask or suggest changes relevant to commercial quality control serum but not to samples of human serum.

Alkaline Phosphatase

Suitability of commercial enzyme control sera for the quality control of activity determinations of L-aspartate aminotransferase and L-alanine aminotransferase in human serum.

Details of a systematic approach to suitability testing of commercial control sera are given for substrate optimized L-aspartate aminotransferase and L-alanine aminotransferase methods at 37 degrees C. Their acceptability for control purposes of standardized methods depends on: (1) the range of control values in relation to borderline values, (2) stability, (3) aspect, clarity, (4) NADH consumption in preincubation time, (5) blank activities, (6) kinetic data as half saturation constants and saturation curves, (7) influence of effectors, (8) isoenzyme pattern. These evaluation criteria are proposed for suitability testing. The term "representativeness" should be introduced as a special criterion for main characteristics of control materials. The authors want to point out the close connection with standardization of methods.

Absorption

Use of equilibrated blood for internal blood-gas quality control.

We have used equilibrated human blood for blood-gas quality control since 1970. In blood equilibrated 24 h after shedding, gas tensions are stable for 4 to 6 h at 0 to 4 degrees C; each control specimen is analyzed several times during that period to resolve malfunctions, etc. Three-fourths of all errors in gas-tension measurement detected with equilibrated blood were detected with the highest-tension controls. Equilibrated blood controls signal about one error every 14 d on each instrument. For more complete quality control, we supplement analysis of equilibrated blood with other sorts of controls, comparing results obtained by assaying each patient's specimen on two instruments being our most effective adjunct. Such comparisons have identified erroneous assays in 3.9% of the specimens tested. The magnitude of interinstrument discrepancies (random errors) have ranged from 9 to 100% of the appropriate determinations. We use control data derived from equilibrated blood analysis for special management purposes (evaluating instruments, quantitating micro- vs. macro-sampling discrepancies, and decreasing instrument-repair costs).

Acidosis

Evaluation of ampouled tonometered buffer solutions as a quality-control system for pH, pCO2, and pO2 measurement.

In response to the need for an adequate quality-control system for blood-pH and blood-gas analyzers, we investigated the practical application of ampouled phosphate-bicarbonate-chloride solutions tonometered with mixtures of carbon dioxide, oxygen, and nitrogen. This system offers three discrete sets of pH, pCO2, AND PO2 values, which are consistent with normal and pathophysiologically high and low values. The stated values were based on the U.S. National Bureau of Standards scale for pH and on gas analysis for pCO2 and pO2. Influence of temperature, air contact, calibration gas, and storage was established. Internal and external quality control by means of these ampoules is presented. The system is stable, accurate, precise, and suitable for simultaneous quality control of pH, pCO2, and pO2 measurements.

Bicarbonates

Regional quality control survey of blood-gas analysis.

We undertook an external quality control survey of blood-gas analysis in 16 laboratories at 13 hospitals. All samples were prepared in the laboratories under investigation by equilibration of blood or serum with gas mixtures of known composition. pH of serum was measured with no significant bias but with an SD of random error 0.026 pH units, which was almost twice the SD of the reference range (0.015). An acceptable random error (half SD of reference range) was not obtained in a longitudinal internal quality control suvey although there were acceptable results for buffer pH in both field and internal surveys. Blood PO2 was measured with no significant bias but with SD of random error 1.38 kPa which reduced to 0.72 kPa by excluding one egregious result. The latter value was just over half of the SD of the reference range (1.2 kPa). PCO2 of blood was also measured without significant bias but with a much smaller SD of random error of 0.28 kPa (by excluding one egregious result), which was again just over half the SD of the reference range (0.51 kPa). Measurements of blood PO2 and PCO2 seem generally acceptable in relation to their respective reference ranges but measurements of pH were unsatisfactory in both internal and external trials.

Blood Gas Analysis

plinkQC: an integrated tool for ancestry inference, sample selection, and quality control in population genetics.

MOTIVATION: Population genetic analyses rely on high quality datasets that pass rigorous controls for sample and marker quality. Many analyses also require additional processing including identification of ancestry and sample relatedness. A software package that addresses all these common, yet crucial tasks is missing. RESULTS: We have developed plinkQC, an R/CRAN package that combines these functionalities into a single software package with detailed vignettes for example applications. plinkQC determines the ancestry of study samples via a pre-trained random forest classifier that reaches 98% performance accuracy with just 5% of marker overlap between reference and user data. To obtain the maximal set of unrelated study samples, we developed a graph-based pruning method, taking both relationship estimates and sample quality into account. We demonstrate optimal sample selection on the 1000 Genomes project, where we retain an additional 71 samples compared to publicly available exclusion lists. Finally, plinkQC bundles these results together with per-individual and per-marker quality control checks into three simple functions and returns both the quality controlled dataset and quality control report about each step of the analysis. AVAILABILITY AND IMPLEMENTATION: plinkQC is available as an R/CRAN package. The documentation and code are available on github: https://meyer-lab-cshl.github.io/plinkQC/ and https://github.com/meyer-lab-cshl/plinkQC_manuscript.

Software

A stable blood product for pH-blood-gas quality control.

We describe how to prepare, store, and use a hemolyzed blood product for simultaneous pH, pCO2, and pO2 quality control. Tonometry of the blood product with two oxygen and two carbon dioxide concentrations resulted in consistent and reproducible values during 38 weeks. The resulting pH values were consistent and reproducible, demonstrating the metabolic acid-base stability of the blood product. We conclude that the proper preparation, storage, and use of the product results in consistent, reproducible, and economical quality control for pH, pCO2 and pO2 blood measurements.

Blood Chemical Analysis

Spatially ordered zygotic genome activation fulfills embryo quality control.

Early embryo development features autonomous, maternally driven cell divisions that self-organize the multicellular blastula or blastocyst tissue. Maternal control cedes to the zygote starting with the onset of widespread zygotic genome activation (ZGA), which is essential for subsequent cell fate determination and morphogenesis. Intriguingly, although ZGA onset is highly regulated at the level of the whole embryo, it can be non-homogenous and precisely patterned at the single-cell level. We previously demonstrated a stereotyped spatial and temporal ordering of ZGA in a model vertebrate embryo. Unknown, however, was whether this precise ZGA patterning was required for development. To address this fundamental question, we devised a strategy to spatially control cell divisions that perturb blastula embryo organization. We demonstrate the feasibility of spatially inverting the cell size pattern of embryos and find that these inverted embryos exhibit a flipped pattern of ZGA. Mispatterned ZGA along the animal-vegetal axis triggers embryo apoptosis, revealing that gastrula embryos have a built-in quality control system to sense inappropriate ZGA patterning, including regionalized defects in transcriptional onset. The quality control response is nonautonomous, dependent on an anti-apoptotic signal that suppresses cell death outside the animal hemisphere. These results reveal the requirement of properly patterned ZGA for normal development and the existence of a surveillance system of embryo quality control exquisitely tuned to the spatial and temporal ordering of genome activation and zygotic gene expression.

Animals

Spatially ordered zygotic genome activation fulfills embryo quality control.

Early embryo development features autonomous, maternally-driven cell divisions that self- organize the multicellular blastula or blastocyst tissue. Maternal control cedes to the zygote starting with the onset of widespread zygotic genome activation (ZGA), which is essential for subsequent cell fate determination and morphogenesis. Intriguingly, although the onset of ZGA is highly regulated at the level of an embryo, it can be non-homogenous and precisely patterned at the single-cell level. We previously demonstrated a stereotyped spatial and temporal ordering of ZGA in a model vertebrate embryo. Unknown, however, was whether this precise ZGA patterning was required for development. To address this fundamental question, we devised a strategy to spatially control cell divisions in the embryo that perturb blastula embryo organization. We demonstrate the feasibility of spatially inverting the cell size pattern of embryos and find that these inverted embryos undergo a flipped pattern of ZGA. Mispatterned ZGA along the animal-vegetal axis causes embryo apoptosis, revealing that gastrula embryos have a built-in quality control system to sense inappropriate ZGA patterning, including regional defects in transcriptional onset. The quality control response is non-autonomous which may depend on anti-apoptotic signals that repress cell death outside of the animal hemisphere. These results reveal the requirement of properly patterned ZGA for normal development and the existence of an embryo quality control response exquisitely tuned to the spatial and temporal ordering of genome activation and zygotic gene expression.

Journal Article

Improved quality control of blood acid-base equilibrium using the daily mean of patients' results: a validation study.

The efficiency of the daily mean of patients' results as a quality control tool for acid-base equilibrium has been studied in three one-month periods. Results for the first two periods were obtained in a stable patient population using tonometry in one period, direct measurement of PCO2 in the other; the third period corresponded to another patient population in which PCO2 was determined by direct measurement. These comparisons allowed demonstration that both methodology and patient populations influenced the optimal truncation limits used to exclude extreme values. Furthermore, the efficiency of the daily mean in itself was poor. Using a cusum plot of the differences of each daily mean from the monthly mean of the daily means, however, dramatically improved this efficiency. So modified, quality control using the daily mean of patients' results represents a useful tool, since the whole analytic process, from blood sampling to result, is checked.

Acid-Base Equilibrium

WxS-QC-a quality control pipeline for human germline short-variant Whole-Genome and Whole-Exome cohorts for population-scale analyses.

SUMMARY: Whole-exome (WES) and whole-genome (WGS) sequencing are rapidly becoming preferred methods for population-scale analysis of the human genetic landscape. However, there are currently no standardized quality control (QC) pipelines for human WES and WGS datasets. In this paper, we present WxS-QC, a powerful, scalable, and convenient pipeline for the QC of human germline short-variant WGS and WES cohorts for population-scale analyses. Our pipeline is suitable for both rare-variant discovery and common-variant association studies. It is based on deeply refactored gnomAD v3 and v4 quality control pipelines, contains several methods we have developed de novo, and is aligned with current best practices in WGS/WES germline cohort QC. We provide all methods in a single codebase, aligned to work together and controlled via a single YAML config, with automatic export of resulting graphs and summary tables, excellent performance and scalability, and comprehensive documentation. The pipeline can run in any UNIX-like environment and can efficiently process cohorts of up to 200 000 whole-exome samples, with the potential to handle bigger datasets. AVAILABILITY AND IMPLEMENTATION: The pipeline code is written in Python using the Hail library and is freely available under the BSD-3 license here: https://github.com/wtsi-hgi/wxs-qc. The detailed description of the pipeline is available in the pipeline documentation: https://github.com/wtsi-hgi/wxs-qc/blob/main/README.md. We also provide an open dataset with all required metadata, which is available at https://wxs-qc-data.cog.sanger.ac.uk/wxs-qc_public_dataset_v3.tar. An example of test dataset analysis is available in the supplementary materials.

Humans

Long-term reproducibility of a new pH/blood-gas quality-control system compared to two other procedures.

The long-term precision and stability of a new quality-control system for blood pH and gas measurements are compared to that of tonometered bicarbonate solutions and serum-based preparations. The new system, consisting of gas-equilibrated bicarbonate solutions in glass ampuls, is shown to be as stable as the serum-based preparation, and as reproducible as either of the other methods. The new system, offering three discrete sets of control values, has certain advantages in the simultaneous quality of pH, carbon dioxide tension, and oxygen tension measurements.

Blood

TargetQC: A targeted quality control framework for clinical genomic testing.

Reliable genetic testing depends on accurate assessment of sequencing quality in clinically relevant genomic regions that directly influence variant interpretation. We developed TargetQC, a flexible quality control framework that supports user-defined gene sets, coverage thresholds, and variant sets for evaluating sequencing performance across exome sequencing (ES) and genome sequencing (GS) platforms. TargetQC assesses exon and gene coverage, identifies regions meeting predefined coverage thresholds, evaluates variant detection accuracy, and measures sequencing quality at pathogenic variant sites. We applied TargetQC to the reference sample NA12878 and 665 clinical samples across five ES platforms and one GS platform. ES-VendorB and ES-VendorE achieved the most complete coverage of OMIM coding regions in NA12878, whereas ES-VendorD and ES-VendorE showed the highest coverage compliance in clinical samples. ES-VendorB and GS demonstrated the highest variant detection accuracy. TargetQC provides a practical framework for benchmarking sequencing performance and informing platform selection in clinical genomics.

exome sequencing

Evaluating Selective Quality Control in Mammalian Oogenesis: Evidence and Opportunities.

The formation and maintenance of the finite mammalian ovarian reserve are critical for fertility and species survival. Genetic and developmental studies have uncovered various mechanisms underlying oocyte development and maturation, revealing two curious features of the ovarian germline: (a) The establishment of the follicle reserve involves an initial massive overproduction of oocyte precursors, and (b) the total number of ovulated oocytes across an animal's fertile lifetime is a very small proportion of the initial ovarian reserve. Many have proposed that this indicates the existence of selective quality control to ensure gamete fitness. Here, we review the findings underlying the hypotheses for germline quality control during prepubertal development, homeostatic fertility, and reproductive aging. We evaluate whether the existing evidence base distinguishes the active selection of specific germ cell subsets from neutral dynamics. Throughout, we discuss strategies for applying statistical frameworks to evaluate selection in oogenesis and the implications of neutrality versus selection at various points in oocyte development.

Oogenesis

The glycoprotein quality control factor Malectin promotes coronavirus replication and viral protein biogenesis.

Coronaviruses (CoV) rewire host protein homeostasis (proteostasis) networks through interactions between viral nonstructural proteins (nsps) and host factors to promote infection. With the emergence of SARS-CoV-2, it is imperative to characterize host interactors shared across nsp homologs. Using quantitative proteomics and functional genetic screening, we identify conserved proteostasis interactors of nsp2 and nsp4 that serve pro-viral roles during infection of murine hepatitis virus - a model betacoronavirus. We uncover a glycoprotein quality control factor, Malectin (MLEC), which significantly reduces infectious titers when knocked down. During infection, nsp2 interacts with MLEC-associated proteins and the MLEC-interactome is drastically altered but retains association with the Oligosaccheryltransferase (OST) complex, a crucial component of viral glycoprotein production. MLEC promotes viral protein levels and genome replication through its quality control activity. Lastly, we show MLEC promotes SARS-CoV-2 replication. Our results reveal a role for MLEC in mediating CoV infection and identify a potential target for pan-CoV antivirals.

Biochemistry and Chemical Biology

QCatch: a framework for quality control assessment and analysis of single-cell sequencing data.

MOTIVATION: Single-cell sequencing data analysis requires robust quality control (QC) to mitigate technical artifacts and ensure reliable downstream results. While tools like alevin-fry and simpleaf (and augmented execution context for the alevin-fry), offer flexibility and computational efficiency to process single-cell data, this ecosystem will further benefit from a standardized QC reporting tailored for its outputs. RESULTS: We introduce QCatch, a Python-based command-line tool that generates comprehensive and interactive HTML QC reports designed specifically for single-cell quantification results. Taking the output directory of alevin-fry or simpleaf as the input, QCatch is able to perform essential processing steps, like cell calling, and generate detailed QC reports that contain informative visualizations and statistics, including unique molecular identifier (UMI) count distributions, sequencing saturation estimates, and splicing status information, for QC assurance. Built for seamless integration into downstream analysis workflows, QCatch exports the processed results in a richly-annotated H5AD format file, a widely used data format common among many downstream single-cell data analysis tools. AVAILABILITY AND IMPLEMENTATION: The source code and documentation of QCatch are available on GitHub at https://github.com/COMBINE-lab/QCatch. QCatch can be installed via both Bioconda and PyPI.

Single-Cell Analysis