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Lipids and lipoproteins

Abstract Lipids are essential sources of energy, structural components of cell membranes, and precursors of hormones, vitamins, and bile acids. Some of them are important in the pathogenesis of atherosclerotic cardiovascular diseases (ASCVD). Notably, cholesterol in low-density lipoproteins (LDL cholesterol) plays a causal role and has become a prime therapeutic target for the prevention of ASCVD. Other clinically established measures of lipid and lipoprotein metabolism include…

Carbohydrates

Abstract Background Carbohydrates are widely distributed in plants and animals. They perform numerous functions, ranging from structural components of deoxyribonucleic acid to serving as sources of energy. Glucose is derived from breakdown of carbohydrates in the diet and in body stores. In addition, glucose can be synthesized from protein or triglyceride. Content This chapter describes the chemistry and metabolism of carbohydrates. Carbohydrates in the diet are…

Kidney function tests

Abstract Background The functional unit of the kidney is the nephron. Because glomerular filtration is the initiating phase of all nephron functions, quantitative or qualitative assessment of the glomerular filtration rate (GFR), or some variable that bears a reasonably constant relationship to it, together with assessment of the integrity of the filtration barrier, generally provide the most useful indices to assess the severity and progress of…

Tumor markers

Abstract Background Tumor markers are substances present in and produced by a tumor or produced by the host in response to a tumor. Measured qualitatively or quantitatively by chemical, immunologic, genomic, or proteomic methods, tumor markers can be used to identify the likely presence of a cancer and/or to differentiate a tumor from normal tissue. Tumor markers can contribute to cancer management as screening tests for…

Serum enzymes

Abstract Background Serum enzymes are measured in medical diagnosis to detect injury to a tissue that contains the measured enzyme. Clinical applications have concentrated mostly on enzymes such as creatine kinase, alanine transaminase, aspartate transaminase, alkaline phosphatase, γ-glutamyltransferase, lactate dehydrogenase (LDH), lipase, and (pancreatic) amylase. Content This chapter describes the use of the clinically most important enzymes as preferred markers in various disease states such as…

Amino acids, peptides, and proteins

Abstract Background Amino acids are not only the building blocks of proteins, but they also play diverse roles in the provision of energy and the formation of a number of other important biomolecules, including hormones, neurotransmitters, and signaling molecules. The polymers of amino acids, peptides and proteins, orchestrate and control the vast array of human physiologic and biochemical processes. The catalog of amino acids, peptides, and…

Point-of-care testing

Abstract Background Point-of-care testing (POCT) is essentially any form of laboratory testing that takes place outside of the conventional or central laboratory setting and encompasses a wide variety of locations. Use of POCT has steadily increased over the 40 or so years since its introduction, principally driven by technology developments and changes in health care delivery that are aimed at delivering less costly and more effective…

Automation in the clinical laboratory

Abstract Background Automation has dramatically changed both the analytical and nonanalytical aspects of clinical laboratory operations. Automation of laboratory test procedures began more than 50 years ago, but nonanalytical automation—including conveyor systems, interfaced analyzers, and automated specimen processing and storage—began in earnest in the 1990s. Today there is a wide selection of automation options designed to improve the quality, throughput, and efficiency of laboratory testing. Content…

Cytometry

Abstract Background Cytometry, or “cell measurement,” can describe any process by which individual biologic cells are counted or characterized, whether or not a human observer is involved. An apparatus used in the process is called a cytometer. From the 1950s on, cytometers, nearly all automated to some degree, have replaced microscopy in an increasing number of applications in both clinical and research laboratories. Some chemical assays…

Microfabrication and microfluidics and their application in clinical diagnostics

Abstract Background Microfluidics is a burgeoning area of analytical chemistry that will impact many fields, including clinical diagnostics. The ability to miniaturize and expedite chemistry with smaller volumes presents the possibility for testing with expedited turnaround times at lower cost and possibly in a portable and handheld format. Content This chapter describes the basic concepts necessary to understand microfluidics at a fundamental level. This includes the…

Immunochemical techniques

Abstract Background Immunoassay is a powerful qualitative and quantitative analytical technique used to detect and measure a wide range of clinically important analytes. The extreme sensitivity and specificity of immunoassays have allowed detection and quantitation of analytes present at very low concentrations not easily measured by other analytical techniques. Content This chapter describes the scope of immunologic assays, including the basics of antigen-antibody binding, antibody production,…

Enzyme and rate analysis

Abstract Background Enzymes are biological catalysts that can be used in the diagnosis and monitoring of disease and their remarkable properties make them sensitive indicators of pathologic change. Metabolism can be regarded as an integrated series of enzymatic reactions and some diseases as a derangement of the physiologic pattern of metabolism. Many enzymes exist in multiple forms, and differences in their properties help in differentiating them…

Proteomics

Abstract Background Clinical proteomics has traditionally referred to experiments that attempt to discover novel biomarkers for disease diagnosis, prognosis, or therapeutic management by using tools that measure the abundance of hundreds or thousands of proteins in a single sample. These discovery experiments began with protein electrophoresis, particularly two-dimensional (2D) gel electrophoresis, and have evolved into workflows that rely very heavily on mass spectrometry (MS). Using the…

Development and validation of small molecule analytes by liquid chromatography-tandem mass spectrometry

Abstract Background The application of liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) represents one of the most compelling opportunities for advancements in human health through the combination of reference measurement procedure capabilities, broad chemical coverage, and a rich history in support of drug development from the 1990s onward. Clinical application of these technologies has begun to gather pace in many laboratories, with diverse applications ranging…

MALDI-TOF mass spectrometry applications in infectious diseases

Abstract Background Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a powerful tool in the clinical microbiology laboratory, enabling accurate identification of bacteria, fungi, and mycobacteria grown in culture. First adopted in some European microbiology laboratories, its ease of use, accuracy, rapid turnaround time, and low cost have led to it becoming standard of care in clinical microbiology laboratories around the world. Content This chapter…

Sample preparation for mass spectrometry

Abstract Background Biological samples such as blood, plasma, serum, and urine contain contaminants that are not suitable for direct analysis and must be removed before chromatographic separation and detection by mass spectrometry. This chapter discusses the classic sample preparation techniques performed in drug analysis laboratories in clinical and research settings and introduces high-throughput applications for improved efficiency. Content The general techniques discussed are dilution, centrifugation, sonication,…

Mass spectrometry

Abstract Background Mass spectrometry (MS) is a powerful analytical technique used to identify and quantify analytes using the mass-to-charge ratio (m/z) of ions generated from a sample. It is useful for the analysis of a wide range of clinically relevant analytes, including small molecules, proteins, and peptides. When MS is coupled with gas or liquid chromatography (GC or LC), the resultant analyzers have expanded analytical capabilities…

Chromatography

Abstract Background Clinical tests often involve the use of one or more steps to isolate, enrich, or separate a target compound from other chemicals in the sample. Chromatography is one of the most common methods for achieving this type of separation. In this method, the components of a mixture are separated based on their differential interactions with two chemical or physical phases: a mobile phase and…

Electrophoresis

Abstract Background Developments in DNA testing, improvements in ease of performance through automation, and advantages of speed and miniaturization afforded by the technique of capillary electrophoresis (CE) have led to a renaissance and growth of electrophoresis as an analytical tool that is widely used in clinical laboratories. These developments and improvements have enabled clinical laboratories to keep pace with higher volumes of testing and to introduce…

Electrochemistry and chemical sensors

Abstract Background Chemical sensors utilizing electrochemical and optical detection methods have become routine analytical tools in clinical chemistry applications, especially for measurement of critical care analytes (blood gases, electrolytes, metabolites) directly in whole blood at point of care or near patient testing. Coupling electrochemical or optical transducers together with chemical or biological recognition elements has expanded the role of chemical sensors for measurement of analytes without…