Validation of the biochemical method for determining the total cholesterol level in the samples of biological fluids as a base for providing the quality of dyslipidemia diagnostics

The main principles of creating a quality system in modern laboratory diagnostics are: standardization of laboratory processes by developing standard operating procedures; general quality management of laboratory research based on the development and implementation of the requirements of international standards (according to ISO 15189: 2015 "Medical laboratories. Basic requirements for quality and competence"); quality control of all stages of the laboratory process through the implementation of the validation procedure. Aim. To develop a methodology for conducting validation procedures to assess the suitability of a biochemical method for determining the level of total cholesterol in biological fluids in the Clinical Diagnostic Laboratory of the Clinical Diagnostic Center of the NUPh. Materials and methods. The object of the study was a standardized method for determining the concentration of total cholesterol. The method was validated using the “Cholesterol Reagent Set” test kit and the standard sample “Chemical control. Reagent kit. Level 1 ", manufactured by High Technology, Inc. (USA) with known concentration. The measurements were carried out on an Express Plus automatic biochemical analyzer manufactured by Bayer Corporation, Germany. When processing the research results, descriptive statistics were used and a number of statistical evaluations were carried out. Results. A protocol and a validation report were developed at the Clinical Diagnostic Laboratory of the CDC NUPh to assess the suitability of the method for determining the concentration of cholesterol in biological fluids by the photometric enzymatic method on an automatic biochemical analyzer Express Plus (using reagents and control material manufactured by High Technology, Inc., USA). The validation characteristics of the method were determined: repeatability and reproducibility, correctness and uncertainty of measurements. Evaluation of the internal laboratory repeatability and reproducibility of this technique indicates the absence of gross errors in the operation of the instrument and statistically important differences in measurements. The assessment of the correctness of the method (carried out using the control material) proved that the systematic error is not significant (according to a given acceptance criterion). The expanded uncertainty calculation showed that the obtained values of the total cholesterol level can be considered accurate and reliable. Conclusions. Validation of the method for determining total cholesterol in human blood by the photometric enzymatic method has proven that this method has performance characteristics that correspond to the regulated ones, meets the established criteria, and the parameters measured with it correspond to the proper ones. Key words: validation, determination method, total cholesterol, repeatability and reproducibility, correctness and uncertainty of measurements

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Improved HPLC Determination of Plasma and Urine Oxalate in the Clinical Diagnostic Laboratory

Journal of Liquid Chromatography ◽

10.1080/10826079208017187 ◽

1992 ◽

Vol 15 (3) ◽

pp. 501-511 ◽

Cited By ~ 11

Author(s):

A. Brega ◽

A. Quadri ◽

P. Villa ◽

P. Prandini ◽

Ji-Qing Wei ◽

...

Keyword(s):

Hplc Determination ◽

Clinical Diagnostic Laboratory ◽

Diagnostic Laboratory ◽

Clinical Diagnostic ◽

Urine Oxalate

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Detecting Antimicrobial Resistance

Tutorial Topics in Infection for the Combined Infection Training Programme ◽

10.1093/oso/9780198801740.003.0056 ◽

2019 ◽

Author(s):

Lynette Phee ◽

David Wareham

Keyword(s):

Phenotypic Expression ◽

Ease Of Use ◽

Clinical Diagnostic Laboratory ◽

Diagnostic Laboratory ◽

Clinical Diagnostic ◽

Control And Prevention ◽

Antimicrobial Susceptibility Tests ◽

Susceptibility Tests ◽

Phenotypic Methods ◽

Cost Of Materials

● To optimize antimicrobial therapy for the management of individual patient’s infection. ● For surveillance purposes, which in turn inform local/national/international clinical guidelines. ● For the management of infection control and prevention. Broadly speaking, resistance is detected by observing its phenotypic expression (activity of the candidate drug(s) against the target bacterium) or detecting the underlying genotypic determinant (resistance genes). Commonly used methods in clinical diagnostic laboratories generally fall under the ‘phenotypic’ category. These share similar traits— ease of use, reproducibility, scalability, quick turnaround of results and relative low cost of materials/reagents required. Moreover, decades of experience and fine-tuning have seen them established as methods of choice in most microbiology laboratories. Most phenotypic test methods are reliant on the use of clinical breakpoints set by national and international bodies (e.g. EUCAST and CLSI) to determine susceptibility/resistance. These guidelines are regularly subject to updates with input from leading experts and latest research findings. It is important for clinical diagnostic laboratories to adhere to best practice guidance set out by these bodies and keep up-to-date with the latest guidelines. Growth characteristics (on artificial media) of the bacterium of interest are extremely important in conventional phenotypic methods. As this presents a big obstacle for slow growers and ‘unculturable’ pathogens (e.g. Mycobacterium tuberculosis, Mycoplasma spp.) it has led to the introduction of genotypic methods of resistance detection in the clinical diagnostic laboratory. meteoric rise in the world of microbiology. Compared with conventional phenotypic methods, molecular genotypic-based tests are better suited for automation and reduce dependence on skilled workers for result interpretation. They therefore deliver the rapid turnaround demanded by modern medicine. Antimicrobial susceptibility tests (ASTs) is a term used to describe a range of phenotypic methods that employ direct observation of the action of antimicrobials against a target microorganism. This is the most commonly used method in clinical diagnostic laboratories for detecting resistance in bacteria. A. Disc diffusion Growth medium: Standardized agar plates (usually unsupplemented, but addition(s) may be necessary for bacteria with specific growth requirements). Antibacterial component: Fixed dose in standard size circular paper discs or tablets.

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Pharmacogenomics and Pharmacogenetics: Future Role of Molecular Diagnostics in the Clinical Diagnostic Laboratory

Clinical Chemistry ◽

10.1373/clinchem.2004.031625 ◽

2004 ◽

Vol 50 (9) ◽

pp. 1526-1527 ◽

Cited By ~ 11

Author(s):

Ralph K Ito ◽

Laurence M Demers

Keyword(s):

Molecular Diagnostics ◽

Clinical Diagnostic Laboratory ◽

Diagnostic Laboratory ◽

Future Role ◽

Clinical Diagnostic

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Real-time reverse transcription PCR (qRT-PCR) and its potential use in clinical diagnosis

Clinical Science ◽

10.1042/cs20050086 ◽

2005 ◽

Vol 109 (4) ◽

pp. 365-379 ◽

Cited By ~ 240

Author(s):

Stephen A. Bustin ◽

Reinhold Mueller

Keyword(s):

Real Time ◽

Reverse Transcription ◽

Therapy Monitoring ◽

Clinical Diagnostics ◽

Clinical Diagnostic Laboratory ◽

Diagnostic Laboratory ◽

Qrt Pcr ◽

Reverse Transcription Pcr ◽

Clinical Diagnostic ◽

Pcr Assays

qRT-PCR (real-time reverse transcription-PCR) has become the benchmark for the detection and quantification of RNA targets and is being utilized increasingly in novel clinical diagnostic assays. Quantitative results obtained by this technology are not only more informative than qualitative data, but simplify assay standardization and quality management. qRT-PCR assays are most established for the detection of viral load and therapy monitoring, and the development of SARS (severe acute respiratory syndrome)-associated coronavirus qRT-PCR assays provide a textbook example of the value of this technology for clinical diagnostics. The widespread use of qRT-PCR assays for diagnosis and the detection of disease-specific prognostic markers in leukaemia patients provide further examples of their usefulness. Their value for the detection of disease-associated mRNA expressed by circulating tumour cells in patients with solid malignancies is far less apparent, and the clinical significance of results obtained from such tests remains unclear. This is because of conceptual reservations as well as technical limitations that can interfere with the diagnostic specificity of qRT-PCR assays. Therefore, although it is evident that qRT-PCR assay has become a useful and important technology in the clinical diagnostic laboratory, it must be used appropriately and it is essential to be aware of its limitations if it is to fulfil its potential.

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The value of the acidified glycerol lysis test with a graphical determination for screening of hereditary spherocytosis

Translational Medicine ◽

10.18705/2311-4495-2019-6-6-51-59 ◽

2020 ◽

Vol 6 (6) ◽

pp. 51-59

Author(s):

T. T. Asatryan ◽

L. B. Gaikovaya ◽

V. V. Slepisheva

Keyword(s):

Hemolytic Anemia ◽

Reliable Method ◽

Hereditary Spherocytosis ◽

Search Methods ◽

Common Cause ◽

Clinical Diagnostic ◽

Biochemical Analyzer ◽

Automatic Biochemical Analyzer

Hereditary spherocytosis (HS) is the most common cause of hemolytic anemia of varying severity can lead to complications such as cholelithiasis, sepsis, etc. Difficulties in diagnosis are asymptomatic and atypical cases HS. It is important to search methods that can detect microspherocytic anemia.The article describes the technique of acidified glycerol lysis test (AGLT), the results and analytical characteristics of the method. Presents a refined acidified glycerol lysis test (AGLT) with incubated blood performed on the automatic biochemical analyzer with a graphical check of the lysis of erythrocytes is a sensitive, fast and reliable method of identification of HS and can be used in clinical diagnostic laboratories.

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On possibility of using routine biochemical techniques for research of alternative biological human liquids

Medical alphabet ◽

10.33667/2078-5631-2019-4-35(410)-54-57 ◽

2020 ◽

Vol 4 (35) ◽

pp. 54-57

Author(s):

S. I. Murskiy

Keyword(s):

Russian Federation ◽

Clinical Laboratory ◽

Biological Fluid ◽

Biological Fluids ◽

Reproductive Age ◽

Laboratory Research ◽

Iso 15189 ◽

Biochemical Analyzer ◽

Automatic Biochemical Analyzer ◽

The Russian Federation

Biochemical studies of alternative human biological fluids are not common in laboratory practice due to the lack of validation methods. The aim of the study was to validatе on semen plasma routine biochemical research methods using an automatic biochemical analyzer. The studies were carried out on a Cobas Integra 400 plus automatic biochemical analyzer (Roche, Switzerland) using original Roche reagents (Switzerland). Materials for the study were 30 samples of semen plasma obtained from clinically healthy men of reproductive age. Planning and organization of validation activities were carried out in accordance with GOST ISO 15189–2015. The functional characteristics of the methods were determined in accordance with Order 45 of the Ministry of Health of the Russian Federation «On a system of measures to improve the quality of clinical laboratory research in healthcare facilities of the Russian Federation», and protocols of the CLSI Institute of Clinical Laboratory Standards (USA). It was possible to establish acceptable levels of linearity, precision and specificity for 31 of the 33 analytes studied. Also, the necessary dilution for 12 analytes was established empirically. Establishing the analytical reliability of biochemical methods for studying semen plasma opens up new possibilities for studying the metabolic characteristics of this biological fluid, and therefore expands the range of possibilities for its use as an object of laboratory research.

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