Urine test
A urine test, also known as urinalysis, is a common laboratory examination of urine that involves physical, chemical, and microscopic analysis to assess overall health and detect various medical conditions.[1] It has been one of the oldest diagnostic tools in medicine, with roots tracing back thousands of years to ancient practices for evaluating bodily health through urine characteristics.[2] Primarily used to identify urinary tract infections (UTIs), kidney and liver diseases, diabetes, and metabolic disorders, the test also aids in routine health screenings, pregnancy monitoring, pre-surgical evaluations, and tracking chronic conditions.[3] The procedure typically begins with urine sample collection, often via the clean-catch midstream method to minimize contamination, where the genital area is cleansed before capturing the middle portion of the urine stream (about 30-60 mL).[1] Analysis proceeds in three main stages: a visual examination evaluates color, clarity, and odor—normal urine is pale yellow and clear, while abnormalities like cloudiness may indicate infection or blood suggesting hematuria; a chemical analysis uses dipstick strips to measure pH, specific gravity, protein, glucose, ketones, bilirubin, nitrites, and leukocyte esterase, helping detect issues such as dehydration, diabetes (via glucose or ketones), or inflammation; and a microscopic examination inspects sediment for cells (red/white blood cells), bacteria, casts, crystals, or other elements that could signal kidney damage or infection.[3][1] In clinical practice, results are interpreted in context with patient symptoms and history, as no single finding confirms a diagnosis; for instance, proteinuria might point to kidney disease but requires further tests for verification.[1] Additional specialized tests, like urine culture for bacterial identification in suspected UTIs, may follow if initial urinalysis suggests infection.[3] Overall, urinalysis remains a cost-effective, non-invasive first-line diagnostic tool, providing rapid insights into renal, metabolic, and systemic health with high clinical utility.[2]Overview
Definition and Purpose
A urine test, also known as urinalysis, is a laboratory examination of urine samples to evaluate physical characteristics, chemical composition, and microscopic elements, providing insights into overall health and specific medical conditions.[4] Urine serves as a non-invasive biofluid that reflects physiological states through the excretion of metabolites, cells, casts, crystals, and potential pathogens, offering a window into renal function, metabolic processes, and systemic health without the need for more invasive procedures.[5][4] The primary purposes of urine tests include screening for urinary tract infections, assessing kidney and liver function, detecting metabolic disorders such as diabetes, monitoring chronic conditions, and identifying markers of drug use or pregnancy.[1][5] These tests are commonly employed in routine health checkups, preoperative evaluations, and diagnostic workups for symptoms like abdominal pain or urinary changes, enabling early detection of issues such as kidney disease, liver problems, or blood disorders.[1][4] Urine tests generally encompass three main categories: physical analysis of appearance (such as color and clarity), chemical evaluation of substances like proteins, glucose, and pH, and microscopic examination for cellular and particulate matter.[1][5] This multifaceted approach allows for a comprehensive yet accessible assessment of health indicators.[4]Historical Background
The practice of urine examination, known as uroscopy, originated over 6,000 years ago in ancient civilizations such as Egypt and Babylon, where physicians visually inspected urine for color, clarity, and sediment to diagnose illnesses.[6] In ancient Greece, Hippocrates (c. 460–377 BC) advanced this approach by systematically using urine analysis to assess bodily humors and organ function, including noting the sweet taste of urine as an indicator of diabetes mellitus.[7] During the medieval period, uroscopy evolved into a refined diagnostic art in Europe and the Islamic world, with physicians like Avicenna (980–1037 AD) describing detailed charts correlating urine characteristics—such as odor and consistency—with specific diseases; tasting was often employed by some physicians to detect sweetness in diabetic cases.[8] In the 19th century, urine testing transitioned from observational methods to chemical analysis, marking a pivotal shift toward scientific precision. British physician Richard Bright, in his 1827 publication Reports of Medical Cases, first systematically linked proteinuria—detected by heating urine samples—with kidney disease, establishing a foundational connection between urinary findings and renal pathology that influenced nephrology's development.[9] Concurrently, advancements in reagents enabled targeted detection of urinary abnormalities; German chemist Hermann von Fehling introduced his test in 1848, using alkaline copper sulfate to quantitatively identify reducing sugars like glucose in urine, revolutionizing diabetes diagnosis.[10] The 20th century brought technological innovations that standardized and automated urine testing for widespread clinical use. In the 1950s, the development of dipstick tests, such as the 1956 Clinistix by the Ames Company (now Bayer Diagnostics), allowed rapid, multi-parameter chemical screening for glucose, protein, and other analytes via enzyme-impregnated strips, dramatically improving accessibility and efficiency over manual methods.[11] By the 1970s, automated analyzers like the Arkray AUTION ANALYZER UA-6 (introduced in 1972) integrated spectrophotometric and flow-based technologies for high-throughput urinalysis, facilitating routine incorporation into laboratory workflows and enhancing diagnostic reliability.[12] These milestones transformed urine testing from an artisanal practice into a cornerstone of modern laboratory medicine.Specimen Collection
Collection Methods
Urine specimens for testing are primarily collected through non-invasive or invasive methods to ensure sample integrity and minimize contamination. Non-invasive techniques, such as spontaneous voiding, are the most common and involve the patient providing a sample directly into a collection device. Invasive methods, including catheterization, are used when patients cannot void voluntarily or when a sterile sample is required.[6] Common types of urine samples include random (spot) samples, which can be collected at any time for routine screening; first morning samples, which are more concentrated and preferred for detecting substances like proteins or hormones; timed collections, such as 24-hour samples for quantitative analysis of analytes like creatinine or electrolytes; and clean-catch midstream samples to reduce external contamination.[1][13][6] The clean-catch midstream technique is widely recommended for routine urinalysis to obtain a representative sample. For individuals with a vagina, the procedure begins with washing hands, sitting on the toilet with legs spread, spreading the labia, and cleaning the urethral area front to back using provided sterile wipes; a small amount of urine is then voided into the toilet, the flow stopped, and the midstream urine collected in a sterile container until halfway full, followed by completing urination in the toilet. For those with a penis, hands are washed, the penis head is cleaned with a sterile wipe (retracting the foreskin if uncircumcised), initial urine is discarded into the toilet, and midstream urine is collected similarly. Samples should be collected after the bladder has held urine for at least 2-3 hours to ensure adequacy.[14][5][1] For 24-hour collections, patients discard the first morning void and collect all subsequent urine over the next 24 hours, including the following morning's first void, into a provided container, often refrigerated during the process to maintain stability.[13] Tools for collection typically include sterile, leak-proof plastic cups or containers supplied by the laboratory, which are wide-mouthed and labeled for identification. In cases where voluntary voiding is not possible, such as in infants or immobilized patients, non-invasive options for infants include applying a sterile adhesive collection bag (U-bag) to the perineal area after cleaning the skin to capture urine during spontaneous voiding, which is preferred to reduce infection risks. If bags are unsuccessful or contamination is a concern, a catheter—a thin, flexible tube—is inserted into the urethra by a healthcare provider to drain urine directly into a sterile receptacle. Suprapubic aspiration, involving needle puncture of the bladder, is rarely used but may be employed for the most sterile samples in neonates or when contamination persists.[6][5][15] Patient preparation is essential to avoid artifacts in the sample. Adequate hydration is advised to facilitate voiding, but excessive fluid intake should be avoided as it may dilute the urine; providers often recommend normal fluid consumption unless specified otherwise. Timing relative to meals, medications, vitamins, or supplements is critical, as certain substances can alter results—patients should inform providers of all intakes, and specific tests may require avoiding interfering agents like caffeine or certain drugs for 8-12 hours prior. For clean-catch methods, patients should avoid menstruation or vaginal discharge to prevent contamination.[5][1][13]Handling and Preservation
Proper handling and preservation of urine specimens are essential to prevent degradation, bacterial proliferation, and alterations in analyte concentrations that could compromise test accuracy. Immediately after collection, urine samples intended for urinalysis should be analyzed within two hours if maintained at room temperature, as delays can lead to increased pH, cell lysis, and bacterial overgrowth.[6] If analysis is delayed, refrigeration at 2°C to 8°C is recommended to slow bacterial growth and preserve cellular elements, with samples remaining viable for up to 24 hours under these conditions.[1][16] For extended storage or specific tests, preservatives are employed to stabilize the sample. Refrigeration alone suffices for routine urinalysis, but chemical additives such as boric acid (typically 10 g per 24-hour collection) inhibit bacterial growth without interfering with most chemical analyses.[17] For tests sensitive to pH changes, such as those measuring catecholamines or metanephrines, acidification with 6N hydrochloric acid (HCl) to achieve a pH below 3 is required to prevent oxidation and degradation of these analytes.[16][17] Sodium azide may be used in commercial collection tubes for certain microbiological or chemical assays to suppress microbial activity.[18] Transport guidelines emphasize maintaining the cold chain and adhering to time constraints to ensure sample integrity. Without preservatives, specimens should be transported and analyzed within 24 hours, using insulated containers with frozen gel packs if ambient temperatures exceed 25°C.[16] Proper labeling, including patient identifiers, collection date and time, and test type, is mandatory, while forensic or drug testing samples require documented chain of custody to prevent tampering.[19] For 24-hour collections, the entire volume must be kept refrigerated or on ice throughout the process and delivered promptly to the laboratory.[20] Common errors in handling can significantly impact results, such as allowing samples to remain at room temperature, which promotes bacterial overgrowth and shifts in pH that falsely elevate glucose or protein levels.[21] Freezing urine is generally discouraged, as it causes cell rupture and artifactual changes in sediment morphology, rendering microscopic examination unreliable.[21] Inadequate mixing before aliquoting or using expired preservatives can also lead to inaccurate quantitative results, underscoring the need for adherence to standardized protocols.[16]Types of Analysis
Chemical Analysis
Chemical analysis of urine, also known as biochemical urinalysis, evaluates the chemical composition and physical characteristics of urine to detect abnormalities indicative of various health conditions. This process typically involves both qualitative and quantitative assessments, with dipstick urinalysis serving as the primary screening method due to its rapid, point-of-care applicability. Dipsticks are reagent-impregnated strips that change color in response to specific urine components, providing semi-quantitative results for multiple analytes simultaneously.[6] More precise quantitative methods, such as colorimetric assays and spectrometry, are employed when detailed measurements are required, particularly for assessing renal function.[6] Dipstick urinalysis commonly tests for several key parameters. The pH level, which measures urine acidity, normally ranges from 4.5 to 8.0, with values around 5.5 to 6.5 being typical; alkaline pH may suggest urinary tract infections or stale samples, while acidic pH can indicate dehydration or diabetic ketoacidosis.[6] Glucose is detected using the glucose oxidase reaction, where its presence (normally negative) signals possible diabetes mellitus, though ascorbic acid can cause false negatives.[6] Protein detection relies on dye-binding methods like tetrabromophenol blue, with normal levels at or below 150 mg per day; elevated protein often points to glomerular or tubular kidney damage, and alkaline urine may lead to false positives.[6] Ketones, absent in healthy urine, are identified through nitroprusside reactions and indicate conditions like diabetic ketoacidosis or starvation, with acidic urine potentially causing false positives.[6] Additional dipstick parameters include bilirubin, which is normally absent and its detection suggests liver dysfunction; urobilinogen, normally 0.1 to 1 mg/dL in random samples or up to 4 mg per day, with elevations linked to hemolysis; nitrites, normally negative and positive in infections from nitrate-reducing bacteria like Escherichia coli, though vitamin C can interfere; leukocytes via esterase activity, normally negative and indicative of inflammation or infection; and blood, normally ≤5 red blood cells per mL, detecting hematuria or hemoglobinuria, with menstrual contamination as a common false positive.[6] Specific gravity, reflecting urine concentration and kidney concentrating ability, typically ranges from 1.002 to 1.035; values outside this may indicate dehydration or renal impairment, and radiographic contrast media can falsely elevate readings.[6] Physical properties assessed during chemical analysis provide initial clues to urine composition. Urine color is normally pale yellow to amber due to urochrome; red or brown hues often result from hematuria or myoglobinuria, while dietary factors like beets can mimic this.[1] Odor is typically mild and "urinoid," but an ammonia-like smell suggests bacterial infection or urea breakdown.[6] Turbidity, or clarity, is normally clear; cloudiness may arise from phosphates, bacteria, or white blood cells, warranting further investigation.[1] Quantitative methods extend beyond dipsticks for accurate analyte measurement. Colorimetric assays and spectrometry quantify substances like proteins (e.g., albumin excretion ≤29 mg/g creatinine indicates normality) and are essential for confirming dipstick findings.[6] A key example is creatinine clearance, which estimates glomerular filtration rate using the formula C = \frac{U_{Cr} \times V}{P_{Cr}}, where U_{Cr} is urine creatinine concentration, V is urine volume per unit time, and P_{Cr} is plasma creatinine concentration; this 24-hour collection method helps diagnose chronic kidney disease.[22] Various artifacts can compromise chemical analysis results. Ascorbic acid (vitamin C) interferes with glucose, nitrite, and bilirubin tests by causing false negatives through its reducing properties.[6] Other interferences include highly pigmented urine affecting color-based readings, glycosuria masking leukocyte esterase, and improper sample storage leading to pH shifts or bacterial overgrowth. Microscopic examination may confirm chemical findings, such as correlating positive blood on dipstick with red blood cell presence.[6]Microscopic Examination
Microscopic examination of urine sediment provides critical insights into renal and urinary tract pathology by identifying and quantifying cellular and acellular components that may indicate underlying conditions.[6] This analysis is performed after concentrating the urine sample to enhance visibility of particulate matter, distinguishing it from chemical tests that detect dissolved substances.[4] Preparation begins with mixing the urine sample thoroughly to ensure homogeneity, followed by transferring 10-15 mL into a centrifuge tube and spinning at approximately 1,500-3,000 rpm for 3-5 minutes to sediment the solids.[4] The supernatant is decanted, and the sediment is resuspended in a small volume of the remaining urine before being placed on a glass slide for examination under a microscope at low (10x) and high (40x) power fields.[23] Staining, such as with Sternheimer-Malbin dye, may be applied to improve contrast for elements like casts and cells, facilitating differentiation of inflammatory from non-inflammatory leukocytes.[24] Key cellular findings include red blood cells (RBCs), where more than 3 RBCs per high-power field (HPF) signifies microscopic hematuria, often pointing to glomerular or lower urinary tract bleeding.[25] White blood cells (WBCs) are also assessed, with greater than 5 WBCs per HPF indicating pyuria, a hallmark of urinary tract infections (UTIs) or inflammation.[26] Epithelial cells, typically squamous or transitional, appear in low numbers as normal contaminants but increase in cases of contamination or urothelial injury.[4] Acellular elements like casts and crystals are vital for diagnosis. Casts form in renal tubules and include hyaline casts, which are normal in small quantities (0-2 per low-power field [LPF]) during dehydration or exercise but may elevate in renal disease.[6] Granular casts, derived from degenerated cellular casts, suggest tubular damage and are associated with acute kidney injury or chronic renal conditions.[27] Crystals vary by urine pH and composition; for instance, uric acid crystals (rhombic or rosette-shaped) are linked to hyperuricemia in gout and predispose to uric acid nephrolithiasis.[28] Quantification standardizes reporting: cells such as RBCs and WBCs are counted per HPF, while casts and larger crystals are reported per LPF to account for field size differences.[29] Clinically, the presence of specific casts correlates with tubular epithelial injury, as granular or cellular casts reflect proteinaceous debris from damaged nephrons.[30] Crystals like uric acid not only signal metabolic disorders such as gout but also indicate risk for calculi formation due to urine supersaturation.[31]Microbiological Culture
The microbiological culture of urine involves the inoculation and incubation of a urine specimen on selective and non-selective media to detect and quantify viable microorganisms, primarily to diagnose urinary tract infections (UTIs).[32] This method allows for the isolation of bacterial pathogens, determination of their quantity, and assessment of antimicrobial susceptibility, guiding targeted therapy.[33] The procedure begins with the quantitative inoculation of the urine sample, typically 1-10 μL, onto agar plates such as 5% sheep blood agar for general growth and MacConkey agar for selective isolation of gram-negative bacteria.[32] The plates are then incubated aerobically at 35-37°C for 24-48 hours to promote colony formation.[32] In some protocols, additional incubation in 5% CO2 may enhance recovery of fastidious organisms.[34] Interpretation relies on colony-forming units per milliliter (CFU/mL), where counts exceeding 10^5 CFU/mL of a single uropathogen in a clean-catch midstream sample are generally considered significant for diagnosing UTI in symptomatic patients.[32] Lower counts, such as 10^2-10^4 CFU/mL, may indicate infection if correlated with symptoms or if multiple pathogens are absent, but counts below 10^5 often suggest contamination rather than true infection.[32] Pathogen identification follows colony morphology observation, using biochemical tests (e.g., API strips or VITEK systems) for metabolic profiling or matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for rapid proteomic analysis, achieving species-level identification in minutes to hours.[35] The most common pathogen isolated is Escherichia coli, accounting for approximately 80% of uncomplicated UTIs, followed by other Enterobacteriaceae such as Klebsiella pneumoniae and Proteus mirabilis, as well as Enterococcus species like Enterococcus faecalis.[36] These gram-negative and gram-positive bacteria predominate due to their uropathogenic traits, including adherence to uroepithelial cells and biofilm formation.[36] Antibiotic susceptibility testing is routinely performed on significant isolates using disk diffusion, broth microdilution, or automated systems to determine minimum inhibitory concentrations, classifying organisms as susceptible, intermediate, or resistant per Clinical and Laboratory Standards Institute guidelines.[32] This step is crucial for managing antimicrobial resistance, particularly in E. coli strains producing extended-spectrum beta-lactamases.[33] Colony count thresholds are adjusted based on collection method to account for potential contamination or reduced bacterial load; for example, a threshold of 10^3 CFU/mL is used for catheterized or suprapubic aspirate specimens, reflecting their lower contamination risk compared to midstream samples.[37] Factors such as delayed processing or prior antibiotic exposure can suppress growth, necessitating prompt plating within two hours of collection or refrigeration if delayed.[32]Specialized Tests
Specialized urine tests encompass advanced assays that detect specific biomarkers, hormones, cellular abnormalities, or molecular signatures not covered by routine analyses, aiding in the diagnosis of conditions such as pregnancy, endocrine tumors, malignancies, infections, and protein disorders. These tests often require specialized laboratory techniques like immunoassays, chromatography, or nucleic acid amplification to achieve high sensitivity and specificity.[38] Hormonal tests in urine primarily target human chorionic gonadotropin (hCG) for pregnancy detection and catecholamines or their metabolites for evaluating adrenal disorders. The hCG urine immunoassay, a qualitative or semi-quantitative chromatographic test, identifies hCG levels exceeding 20 mIU/mL in urine, typically detectable 2-3 days before a missed menstrual period, confirming early pregnancy through monoclonal antibody binding to hCG antigens.[38][39] For pheochromocytoma, a catecholamine-secreting tumor, 24-hour urine collection measures fractionated metanephrines and catecholamines (epinephrine, norepinephrine, dopamine) using high-performance liquid chromatography (HPLC) or liquid chromatography-tandem mass spectrometry (LC-MS/MS), with elevated levels (e.g., metanephrines >2 mg/24 hours) indicating the condition due to their high diagnostic sensitivity of up to 90%.[40][41] These tests require acid preservation of urine to stabilize metabolites and patient adherence to collection protocols avoiding caffeine or certain medications.[42] Urine cytology serves as a non-invasive method for detecting urothelial malignancies, particularly bladder cancer, by examining shed cells for atypia. Specimens, often from voided urine or bladder washings, are centrifuged, smeared onto slides, and stained with the Papanicolaou method to highlight nuclear irregularities, cytoplasmic features, and chromatin patterns indicative of high-grade urothelial carcinoma, achieving sensitivity of 40-60% for detecting atypical or malignant cells.[43][44] This test is especially useful for monitoring recurrence in patients with known bladder cancer, where high-grade lesions show marked nuclear enlargement and hyperchromasia.[45] Molecular tests leverage polymerase chain reaction (PCR) or nucleic acid amplification techniques (NAAT) to identify pathogens or substances in urine with greater precision than culture methods. For sexually transmitted infections, urine-based PCR detects Chlamydia trachomatis and Neisseria gonorrhoeae DNA, offering sensitivity over 90% in first-void urine samples from symptomatic or high-risk individuals, facilitating rapid diagnosis without invasive swabs.[46][47] In drug screening, initial enzyme-linked immunosorbent assay (ELISA) or immunoassay identifies presumptive positives for substances like opioids or amphetamines, followed by gas chromatography-mass spectrometry (GC-MS) confirmation to quantify metabolites (e.g., morphine >2,000 ng/mL) and rule out cross-reactivity, ensuring forensic or workplace accuracy with specificity near 100%.[48][49] Additional specialized assays include urine protein electrophoresis for detecting Bence Jones proteins and nuclear matrix protein 22 (NMP22) for bladder cancer monitoring. Electrophoresis separates urine proteins under an electric field, identifying monoclonal light chains (Bence Jones proteins) in 50-80% of multiple myeloma cases, where their presence in 24-hour collections exceeding 1 g/day signals light-chain disease and requires immunofixation for confirmation.[50][51] The NMP22 assay, an enzyme immunoassay detecting nuclear mitotic apparatus protein fragments from apoptotic tumor cells, yields positive results in urine at levels >10 U/mL, providing 50-70% sensitivity for bladder cancer recurrence when used adjunctively with cystoscopy.[52][53] These tests enhance targeted diagnostics but must be interpreted alongside clinical context to avoid false positives from benign conditions.[54]Interpretation of Results
Normal Reference Ranges
Normal reference ranges for urinalysis provide baseline values for physical, chemical, and microscopic parameters, aiding in the identification of deviations that may warrant further investigation. These ranges can vary slightly between laboratories due to methodological differences, but standard values are widely established in clinical practice.[6]Physical Characteristics
Specific gravity, which measures urine concentration, normally falls between 1.005 and 1.030, indicating adequate kidney function in concentrating or diluting urine relative to plasma.[55] The color of healthy urine is pale yellow to amber, primarily due to the pigment urochrome, though it may appear clearer when dilute.[56] Clarity is normally clear or translucent, while odor is typically aromatic or "urinoid."[6]Chemical Parameters
Urine pH typically ranges from 4.5 to 8.0, reflecting the body's acid-base balance, with an average around 6.0.[6] Standard chemical tests yield negative results for glucose, ketones, blood (hemoglobin), and nitrites in healthy individuals, as these substances are not typically present in detectable amounts.[6] Protein is normally negative or trace on dipstick (typically <30 mg/dL), corresponding to a total daily excretion of less than 150 mg.[6]Microscopic Examination
Under microscopic analysis, red blood cells (RBCs) are present at 0 to 5 per high-power field (HPF), while white blood cells (WBCs) range from 0 to 5 per HPF, indicating no significant inflammation or bleeding.[6] Casts and crystals should be absent or minimal, with hyaline casts up to 5 per low-power field considered normal and no excess of other types like granular or cellular casts.[6]Variations in Reference Ranges
Reference ranges can be influenced by physiological factors such as diet, where high-protein intake acidifies urine (lowering pH), and vegetarian diets may raise pH toward alkaline levels.[6] Hydration status affects specific gravity, with dehydration increasing it above 1.030 and overhydration decreasing it below 1.005.[55] Age and sex generally do not alter ranges substantially in adults, though newborns may exhibit slightly higher pH and protein levels transiently.[6]| Parameter | Normal Range | Source |
|---|---|---|
| pH | 4.5–8.0 | NCBI StatPearls |
| Specific Gravity | 1.005–1.030 | MedlinePlus |
| Color | Pale yellow to amber | MedlinePlus |
| Glucose | Negative | NCBI StatPearls |
| Protein | Negative or trace (dipstick, <30 mg/dL) | NCBI StatPearls |
| Ketones | Negative | NCBI StatPearls |
| Blood | Negative | NCBI StatPearls |
| Nitrites | Negative | NCBI StatPearls |
| RBCs | 0–5/HPF | NCBI StatPearls |
| WBCs | 0–5/HPF | NCBI StatPearls |