ROUTINE URINALYSIS

Routine Urinalysis

The routine urinalysis is a quick and relatively inexpensive test which can be readily performed in a modest clinical laboratory. The results are useful in a variety of situations not limited to those directly involving the urinary tract. Routine urinalysis is an essential part of the diagnostic evaluation of sick patients and the results should be interpreted along with the results of a chemistry panel. Ideally urine should always be collected at the same time as blood for hematology and clinical chemistry AND before any treatment (including intravenous fluids) is administered. Complete interpretation of results of chemistry panels cannot be performed without concurrent knowledge of the urinalysis, particularly if there are abnormalities in renal (e.g. urea nitrogen and creatinine) or acid-base parameters on the chemistry panel. Similarly, interpretation of some abnormalities in urine (e.g. glucosuria, ketonuria) is facilitated by concurrent knowledge of chemistry results.

Required equipment and materials include: microscope, clinical centrifuge, refractometer, and multiple reagent dipsticks. Urine should be examined as soon as possible after collection, because artifacts will occur in the urine over time (cells lyse, crystals form in vitro). If a delay is anticipated before analysis, the urine should be refrigerated. Refrigerated urine should always be brought to room temperature before testing. For more information on the effects of timing of sample collection and analysis, click here.

Routine urinalysis consists of the following steps:

• Visual urine attributes: Volume, color and turbidity.
  It is ideal to standardize the volume of urine from which the urine sediment is prepared. In human medicine, 10 ml of urine is used as the standard volume. This is difficult to accomplish in many animals, particularly small patients, hence we try and standardize the urine volume from which the urinalysis is performed (regardless of the volume received) to 3 ml in the laboratory at Cornell University. However, we often do not receive even this much urine for analysis. Urine volume affects the results of the urine sediment examination, because the semi-quantitative results of the sediment are derived from the standard urine volume and will differ between urine collections of different volumes. Observations of color and turbidity are made on the well-mixed urine specimen.
• Specific gravity: This is a measure of the urine concentrating ability of the animal. The specific gravity should be read on the refractometer using the urine supernatant.
• Dipstick analysis: Dipsticks consist of various pads containing chemical ingredients which provide a color change when a particular analyte is present in urine. This color change is converted to a semi-quantitative result for the analyte in question. In animals, the dipstick is used to give results for pH, protein (mostly albumin), glucose, ketones (primarily acetoacetic acid), bilirubin (primarily the conjugated form) and proteins containing a heme group (a porphyrin ring with iron in its center). There are also dipstick pads for urine specific gravity, nitrate, leukocytes and urobilinogen on commercially available dipsticks. These are either not accurate in animals or do not provide much additional information in animals and are seldom, if ever, reported. The dipstick analysis is usually performed on uncentrifuged urine, unless there is marked hematuria (which may affect interpretation of the color changes on the dipstick). In urines with marked hematuria, the interfering erythrocytes can be sedimented by centrifugation and the dipstick analysis can be performed on the supernatant. When using, follow manufacturer's directions on storage and use of the reagents. The package insert also contains useful information about test limitations and interfering substances.
  In clinical practice, we visually (and subjectively) assess the degree of color change on the dipstick (e.g. 1+, 2+). At Cornell University, we no longer visually examine the multistix to determine the dipstick results. We actually use an instrument, called the Criterion, to "read" the color changes on a urinary dipstick. The machine corrects for the urine color and provides semi-quantitative results (except for heme protein) for the urine parameters. Clickhere for information on how the results provided from this machine correlate to the visual estimates reading the dipstick manually.
• Sediment examination: For this examination, the standard volume of urine is centrifuged in a low speed centrifuge. The supernatant is decanted and the urine is gently resuspended in a standard volume (0.5 ml) of urine supernatant. A drop of the resuspended urine is placed on a slide, coverslipped and examined under a light microscope using the 10x and 40x objectives (this is frequently called a "wet prep"). Subdued lighting is necessary to increase refractility of the unstained urine elements (lower condenser and/or close down the substage iris diaphragm). At Cornell University, we do not use stains (e.g. Sedi-stain) to highlight urinary constituents, we only examine unstained urine sediments. To examine a urine sediment, we do the following:
• Low magnification: Examine the entire coverslip using the 10x objective. At this magnification, casts, large crystals, debris, parasitic ova are semi-quantified.
• High magnification: Specific structures identified at low magnification (e.g. casts) and several random fields are examined using the 40x (high dry) objective. At this higher magnification, leukocytes, erythrocytes, epithelial cells, fat droplets, small crystals, sperm, debris and bacteria are semi-quantified.

Source : © Cornell University