Volume 53, Issue s2 - Special Supplemental Issue: Urinary sediment: still an important diagnostic tool / Guest Editor: Giovanni B. Fogazzi

The naked-eye appearance of the urine must have been studied by shamans and healers since the Stone Age, and an elaborate interpretation of so-called Uroscopy began around 600 AD as a form of divination. A 1000 years later, the first primitive monocular and compound microscopes appeared in the Netherlands, and along with many other objects and liquids, urine was studied from around 1680 onwards as the enlightenment evolved. However, the crude early instruments did not permit fine study because of chromatic and linear/spherical blurring. Only after complex multi-glass lenses which avoided these problems had been made and used in the 1820s in London by Lister, and in Paris by Chevalier and Amici, could urinary microscopy become a practical, clinically useful tool in the 1830s. Clinical urinary microscopy was pioneered by Rayer and his pupils in Paris (especially Vigla), in the late 1830s, and spread to UK and Germany in the 1840s, with detailed descriptions and interpretations of cells and formed elements of the urinary sediment by Nasse, Henle, Robinson and Golding Bird. Classes were held, most notably by Donné in Paris. After another 50 years, optical microscopy had reached its apogee, with magnifications of over 1000 times obtainable free of aberration, using immersion techniques. Atlases of the urinary sediment were published in all major European countries and in the US. Polarised light and phase contrast was used also after 1900 to study urine, and by the early 20th century, photomicroscopy (pioneered by Donné and Daguerre 50 years previously, but then ignored) became usual for teaching and recording. In the 1940s electron microscopy began, followed by detection of specific proteins and cells using immunofluorescent antibodies. All this had been using handheld methodology. Around 1980, machine-assisted observations began, and have dominated progress since.

Examination of the urine under the microscope using polarised light is invaluable for detecting and identifying lipid particles. Attention to the shape of these Maltese cross bearing bodies can distinguish conventional fat particles from Fabry bodies with great sensitivity and specificity across a wide phenotypic spectrum. This could be a cheap and rapid tool for screening subjects suspected of having Fabry disease for renal involvement. It remains to be seen whether there is value in integrating polarised light into automated urine microscopy machines, but potentially this could greatly help the pathologist or nephrologist in identifying unusual urinary particles, and broaden the capacity for larger scale screening.

The morphological analysis of urine sediment is an essential part of urinalysis and casts are important elements of urinary sediment. Their shape is typically cylindrical, with extremities often rounded. Casts form within the renal tubules and are made of Tamm-Horsfall glycoprotein (THG). Under some physiological or pathological conditions THG fibrils aggregate giving rise to casts, whose formation is favoured by a number of factors including high urine osmolality and/or low pH. Casts can be found in normal subjects, in non-renal conditions, such as fever, dehydration, and acute heart failure as well as in virtually all renal diseases. Casts can be classified on the basis of their morphology as hyaline, granular, waxy, fatty, cellular (leukocytic, erythrocytic, epithelial), containing crystals or microorganisms, pigmented and mixed. As the correct identification of casts is crucial for an accurate and timely diagnosis of renal disorders, laboratory professionals should be trained to identify and classify casts properly.

Crystalluria is a marker of urine supersaturation with substances deriving from metabolic disorders, inherited diseases or drugs. The investigation of crystalluria must be done according to a protocol which includes the delivery to the laboratory of a proper urine sample, the use of a microscope equipped with polarized light, the accurate knowledge of urine pH, and a comprehensive examination of the crystals, which is based on their identification, quantification and size measurement. For unusual crystals, infrared spectroscopy may also be needed. The main urinary crystalline categories include: calcium oxalates, calcium phosphates, uric acids and urates, struvite, aminoacids (cystine), purines (2,8-dihydroxyadenine and xanthine) and drugs (e.g. sulfamethoxazole, amoxycillin, ceftriaxone, atazanavir). The investigation of crystalluria is a cheap and valuable tool for the detection and the monitoring of inherited and acquired diseases associated with urinary stone formation or renal function impairment – either acute or chronic – due to intrarenal crystal precipitation.

External quality assessment (EQA) schemes for urinalysis have been provided by Labquality Ltd, the publicly owned EQA service provider in Finland, since the 1980s. In 2014, the scheme on urine particle identification had 329 participating laboratories, out of which 60% from 19 countries were outside Finland. Each of the four annual web-based rounds were distributed with four Sternheimer-stained images from a single patient sample, as viewed both by bright-field and phase-contrast optics. Participants reported classified categories either at the basic or at the advanced level. Participating laboratories received assessment of their analytical performance as compared to their peers, including reflections from clinical data and preanalytical detail of the specimen. In general, reporting of basic urine particles succeeded in the eight schemes during the years 2013–2014 as follows: red blood cells 82%–92%, white blood cells 82%–97%, squamous epithelial cells 92%–98%, casts 84%–94%, and small epithelial cells 73%–83% (minimum and maximum of expected or accepted reports). This basic level of differentiation is used in routine laboratory reports, or as verification of results produced by automated instruments. Considerable effort is needed to standardise national procedures and reporting formats, in order to improve the shown figures internationally. Future technologies may help to alleviate limitations created by single digital images. Despite improvements, degenerating cells and casts always exhibit intermediate forms creating disputable classifications. That is why assessment of performance should encompass justified acceptable categories into the assessed outcomes. Preanalytical and clinical detail provide essential added value to morphological findings.

Background: Manual microscopy still represents the gold standard for urinary sediment (US) examination. We report the results obtained in the period 2012–2015 by the EQA Italian program on US, which today involves about 260 laboratories. Methods: The program includes four surveys per year. In two surveys, participants are asked to supply identification and clinical association of US particles. In two other surveys, they are asked to supply the diagnosis of clinical cases, presented with images, some key laboratory findings and a short clinical history. Sixty-six images of US particles (21 cells, 2 lipids, 21 casts, 10 crystals, 3 microorganisms, 15 contaminants) and seven clinical cases were presented. Results: The correct identification rate for each category of particles, in decreasing order, was: micro-organisms (mean±SD: 92.4%±4.5%), lipids (92.0%±1.8%), casts (82.8%±8.8%), crystals (79.4%±29.8%), cells (77.3%±13.5%), and contaminants (70.9%±22.2%). For 13 particles, a correct clinical association was indicated by 91.5%±11.7% of participants, while it was 52.7% for particles associated with urinary tract infection. For clinical cases, due to a high rate of particles misidentification, only 44.3%±10.1% of participants achieved access to clinical diagnosis, which was then correctly indicated by 92.5%±5.3% of them. Conclusions: The results of the EQA program confirm that, while some US particles are well known in terms of identification, clinical association and clinical meaning, others particles still are not, and this represents an important reason to encourage EQA programs on US.

Background: Urinary tract infections (UTI) are among the most common bacterial infections and urine samples represent a large proportion of the specimens processed in clinical microbiology laboratories, up to 80% of which, however, yield negative results. Automated microscopy is widely used for urine sediment analysis and has recently been evaluated in a few studies for bacteriological screening of urine samples, achieving high levels of performance. Methods: We present a study in which urine samples from both inpatients and outpatients, with either clean-catch or indwelling catheter urine samples, were screened for UTI by urine culture, as the reference method, and the automated urine analyser sediMAX, for the detection of bacteria, leukocytes and yeasts. Results: In total, 3443 urine samples were evaluated. When a single algorithm was adopted for sediMAX to screen the total patient population, 96.4% sensitivity, 75.4% specificity, 57.8% positive predictive value, and 98.4% negative predictive value were found. However, for male outpatients and all patients with indwelling catheter other algorithms were necessary to improve performances. Altogether, with sediMAX false negative rate was 2.4% and false positive rate was 27.6%. In addition, 54% of the investigated samples could have avoided urine culture. Conclusions: After the identification of specific algorithms for different patient subgroups, the automated urine analyser sediMAX can be reliably employed in the screening of UTI.

Automated urinalysis devices are reproducible, accurate and faster than the standard manual microscopy. Economic analysis has shown that decreases in turn-around-time and labour cost savings offered by these devices make them more economic than manual microscopy.