scholarly journals Screening of Bacteria in Urine Using Luciferin-Luciferase Assay of Microbial ATP: A Comparative Study

1982 ◽  
Vol 19 (6) ◽  
pp. 416-420 ◽  
Author(s):  
Aimo Ruokonen ◽  
Matti Koskinen ◽  
Maija Leinonen ◽  
Anja Tiilikainen ◽  
Reijo Vihko
Keyword(s):  
Firefly Luciferase ◽  
Rapid Screening ◽  
Luciferase Assay ◽  
High Power Field ◽  
Urine Sediment ◽  
Culture Techniques ◽  
Colony Forming Units ◽  
Luminescence Assay ◽  
Tract Infections ◽  
Urine Specimens

A total of 3227 urine specimens were analysed to investigate the applicability of a firefly luciferase assay of microbial adenosine triphosphate (ATP) for the rapid screening of bacteriuria in clinical specimens. Urine sediment, dipslide culture, and three plate cultures were also investigated in the majority of the urine specimens, the plate cultures serving as the reference. Of the specimens with positive plate culture (≥105 colony forming units (CFU)), leucocyte content of the spun urine sediment was negative (≤4 cells per high-power field) in 34% and bacterial content was negative in 14% (no bacteria seen microscopically); the dipslide test was negative (<105 CFU) in 16%, and the luminescence assay of ATP in 7% (≥ 500 relative light units). Of the urine specimens forming less than 105 CFU on plate cultures, leucocytes were positive in 15%, spun sediment bacteria in 21%, the dipslide test in 0 · 5%, and the luminescence assay of ATP in 11%. When used as a screening test for further studies by complete culture techniques, the luminescence assay of microbial ATP can improve the rapid diagnosis of urinary tract infections.

Review of Enterococci Isolated from Canine and Feline Urine Specimens from 2006 to 2011

2015 ◽  
Vol 51 (3) ◽  
pp. 148-154 ◽  
Author(s):  
Kate S. KuKanich ◽  
Brian V. Lubbers
Keyword(s):  
Medical Records ◽  
Urine Culture ◽  
Bacterial Species ◽  
Clinical Signs ◽  
Multidrug Resistant ◽  
Urine Sediment ◽  
Colony Forming Units ◽  
Enterococcus Spp ◽  
Tract Infections ◽  
Urine Specimens

Canine and feline urine culture reports and medical records were reviewed at a veterinary teaching hospital from 2006 to 2011 for enterococcal growth, coinfections, antimicrobial resistance, urine sediment findings, clinical signs, and concurrent conditions. Of all of the urine specimens with significantly defined colony-forming units/mL, Enterococcus (E.) faecalis was the only enterococci isolated from cats and predominated (77.4%) in dogs followed by E. faecium (12.9%), E. durans (3.2%), and other Enterococcus spp. (6.5%). The majority of specimens with significant enterococcal growth resulted in complicated urinary tract infections in 83.9% of dogs and 81.8% of cats. Specimens with only enterococcal growth were more common than those mixed with other bacterial species. Cocci were observed in urine sediments of 8 out of 8 cats and 21 out of 25 dogs with available concurrent urinalyses. Pyuria was noted in 5 out of 8 feline and 15 out of 25 canine urine sediments, and pyuria in dogs was associated with growth of only enterococci on aerobic urine culture. Multidrug resistance was identified in 6 out of 11 cats and 7 out of 31 dogs, and E. faecium isolates from dogs were 4.5× more likely to be multidrug resistant than E. faecalis.

scholarly journals Urine Sediment Findings and the Immune Response to Pathologies in Fungal Urinary Tract Infections Caused by Candida spp.

2020 ◽  
Vol 6 (4) ◽  
pp. 245
Author(s):  
José Antonio Tesser Poloni ◽  
Liane Nanci Rotta
Keyword(s):  
Immune Response ◽  
Urinary Tract ◽  
Urine Samples ◽  
Candida Spp ◽  
Urine Sediment ◽  
Culture Techniques ◽  
Detection And Identification ◽  
Identification Of Fungi ◽  
Tract Infections ◽  
Urine Markers

Fungi are pathogenic agents that can also cause disseminated infections involving the kidneys. Besides Candida, other agents like Cryptococcus spp. can cause urinary tract infection (UTI), as well as other non-yeast fungi, especially among immunocompromised patients. The detection and identification of fungi in urine samples (by microscopy and culture) plays an essential role in the diagnosis of fungal UTI. However, variable cutoff definitions and unreliable culture techniques may skew analysis of the incidence and outcome of candiduria. The sediment analysis plays a key role in the identification of fungal UTI because both yeasts and pseudohyphae are easily identified and can be used as a clinical sign of fungal UTI but should not be overinterpreted. Indeed, urine markers of the immune response (leukocytes), urine barriers of tissue protection (epithelial cells), and urine markers of kidney disease (urinary casts) can be found in urine samples. This work explores the manifestations associated with the fungal UTI from the urinalysis perspective, namely the urinary findings and clinical picture of patients with fungal UTI caused by Candida spp., aspects associated with the immune response, and the future perspectives of urinalysis in the diagnosis of this clinical condition.

Rapid screening for bacteriuria with a laser nephelometer

1984 ◽  
Vol 30 (7) ◽  
pp. 927-929
Author(s):  
Paul Bayardelle ◽  
Hervé Richet
Keyword(s):  
Standard Method ◽  
False Positive ◽  
False Positive Rate ◽  
Rapid Screening ◽  
Loop Method ◽  
Colony Forming Units ◽  
Positive Rate ◽  
Significant Bacteriuria ◽  
Urine Specimens

Detection of significant bateriuria with a laser nephelometer was evaluated in this study and compared with the results obtained by the quantitative loop method. We screened 1002 urine specimens and 220 (21.95%) were found to be positive at ≥ 105 colony-forming units (CFU)/mL of urine by the standard method. Of the 220 positive specimens, 210 (95.4%) were detected in 6 h or less and 177 (80.4%) were detected within 3 h. The false-positive rate was 2.3% at 3 h and 19.7% at 6 h. These findings suggest that a 6-h procedure is necessary to detect 95% or more of significant bacteriuria. Laser nephelometer is versatile and can be used for rapid screening of bacteriuria.

scholarly journals Further evaluation of the test for detection of antibody-coated bacteria in urine sediment

1977 ◽  
Vol 5 (5) ◽  
pp. 510-513
Author(s):  
S R Jones ◽  
J Johnson
Keyword(s):  
Immune Response ◽  
Urinary Tract ◽  
Urinary Tract Infections ◽  
Diagnostic Laboratory ◽  
Urine Sediment ◽  
Tract Infections ◽  
Antibody Coating ◽  
Urine Specimens

To assess the reproducibility of the test for detection of antibody-coated bacteria in urine sediment as it might be used in a diagnostic laboratory for classification of urinary tract infections, multiple urine specimens from 83 patients were tested. The results were reproducible and consistent, or if inconsistent potentially explainable, in all but four patients. The explanation for inconsistencies include the immune response to the infecting bacteria, nonspecificity of the antibody coating the bacteria, antibody in prostatic secretions, and antibody-coated bacteria contaminating the urine specimens.

scholarly journals Exploring the possibilities of infrared spectroscopy for urine sediment examination and detection of pathogenic bacteria in urinary tract infections

2020 ◽  
Vol 58 (10) ◽  
pp. 1759-1767
Author(s):  
Mieke Steenbeke ◽  
Sander De Bruyne ◽  
Jerina Boelens ◽  
Matthijs Oyaert ◽  
Griet Glorieux ◽  
...  
Keyword(s):  
Infrared Spectroscopy ◽  
Urinary Tract ◽  
Urinary Tract Infections ◽  
Pathogenic Bacteria ◽  
Agar Plate ◽  
Principal Component ◽  
Blood Agar Plate ◽  
Urine Sediment ◽  
Flight Mass Spectrometry ◽  
Tract Infections

AbstractObjectivesIn this study, the possibilities of Fourier-transformed infrared spectroscopy (FTIR) for analysis of urine sediments and for detection of bacteria causing urinary tract infections (UTIs) were investigated.MethodsDried urine specimens of control subjects and patients presenting with various nephrological and urological conditions were analysed using mid-infrared spectroscopy (4,000–400 cm−1). Urine samples from patients with a UTI were inoculated on a blood agar plate. After drying of the pure bacterial colonies, FTIR was applied and compared with the results obtained by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Chemometric data analysis was used to classify the different species.ResultsDue to the typical molecular assignments of lipids, proteins, nucleic acids and carbohydrates, FTIR was able to identify bacteria and showed promising results in the detection of proteins, lipids, white and red blood cells, as well as in the identification of crystals. Principal component analysis (PCA) allowed to differentiate between Gram-negative and Gram-positive species and soft independent modelling of class analogy (SIMCA) revealed promising classification ratios between the different pathogens.ConclusionsFTIR can be considered as a supplementary method for urine sediment examination and for detection of pathogenic bacteria in UTI.

scholarly journals Affordable Luciferase Reporter Assay for Cell-Based High-Throughput Screening

2012 ◽  
Vol 18 (4) ◽  
pp. 453-461 ◽  
Author(s):  
Ellen Siebring-van Olst ◽  
Christie Vermeulen ◽  
Renee X. de Menezes ◽  
Michael Howell ◽  
Egbert F. Smit ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Firefly Luciferase ◽  
Luciferase Assay ◽  
Luciferase Reporter ◽  
Chemical Components ◽  
Simple Liquid ◽  
Luciferase Reporter Construct ◽  
Reagent Cost ◽  
Firefly Luciferase Gene

The firefly luciferase gene is commonly used in cell-based reporter assays. Convenient luciferase assay reagents for use in high-throughput screening (HTS) are commercially available. However, the high cost of these reagents is not within the means of some academic laboratories. Therefore, we set out to develop an affordable luciferase assay reagent applicable in an HTS format using simple liquid-handling steps. The reagent was homemade from individual chemical components and optimized for luminescence intensity and stability. We determined the minimal concentrations of the most expensive components, dithiothreitol (DTT) and D-luciferin, resulting in a total assay reagent cost of less than 1 cent per sample. Signal stability was maximized by omission of coenzyme A and reduction of DTT concentration. The assay was validated in a high-throughput setting using two cancer cell lines carrying a p53-dependent luciferase reporter construct and siRNAs modulating p53 transcriptional activity. Induction of p53 activity by silencing PPM1D or SYVN1 and reduction of p53 activity by silencing p53 remained constant over a 2-h measurement period, with good assay quality (Z′ factors mostly above 0.5). Hence, the luciferase assay described herein can be used for affordable reporter readout in cell-based HTS.

Urinary Tract Infections in Febrile Infants Younger Than 8 Weeks of Age

PEDIATRICS ◽  
1990 ◽  
Vol 86 (3) ◽  
pp. 363-367 ◽  
Author(s):  
Ellen F. Crain ◽  
Jeffrey C. Gershel
Keyword(s):  
Emergency Department ◽  
Urinary Tract ◽  
Urinary Tract Infections ◽  
Urine Culture ◽  
White Blood Cells ◽  
Laboratory Data ◽  
Pediatric Emergency ◽  
Urine Specimen ◽  
High Power Field ◽  
Tract Infections

In this prospective study of 442 infants younger than 8 weeks of age who attended a pediatric emergency department with temperature ≥100.6°F (38.1° C), urinary tract infections (UTIs) were found in 33 patients (7.5%), 2 of whom were bacteremic. Clinical and laboratory data were not helpful for identifying UTIs. Of the 33 patients with UTIs, 32 had urinalyses recorded; 16 were suggestive of a UTI (more than five white blood cells per high-power field or any bacteria present). Of the 16 infants with apparently normal urinalysis results, three had an emergency department diagnosis suggesting an alternative bacterial focus of infection. If the physician had decided on the basis of apparently normal urinalysis results to forgo obtaining a urine culture, more than half of the UTIs would have been missed. Bag-collected specimens were significantly more likely to yield indeterminate urine culture results than either catheter or suprapublic specimens. In addition, uncircumcised males were significantly more likely to have a UTI than circumcised boys. These results suggest that a suprapubic or catheter-obtained urine specimen for culture is a necessary part of the evaluation of all febrile infants younger than 8 weeks of age, regardless of the urinalysis findings or another focus of presumed bacterial infection.

Febrile Infants at Low Risk for Serious Bacterial Infection—An Appraisal of the Rochester Criteria and Implications for Management

PEDIATRICS ◽  
1994 ◽  
Vol 94 (3) ◽  
pp. 390-396 ◽  
Author(s):  
Julie A. Jaskiewicz ◽  
Carol A. McCarthy ◽  
Amy C. Richardson ◽  
Kathleen C. White ◽  
Donna J. Fisher ◽  
...  
Keyword(s):  
Confidence Interval ◽  
High Power ◽  
Microscopic Examination ◽  
Bacterial Infections ◽  
Antimicrobial Agents ◽  
Low Risk ◽  
High Power Field ◽  
Urine Sediment ◽  
Predictive Values ◽  
Risk Criteria

Objective. Prospective studies were conducted to test the hypothesis that infants unlikely to have serious bacterial infections (SBI) can be accurately identified by low risk criteria. Methods. Febrile infants (rectal T ≥ 38°C) ≤60 days of age were considered at low risk for SBI if they met the following criteria: 1) appear well; 2) were previously healthy; 3) have no focal infection; 4) have WBC count 5.0-15.0 x 109 cells/L (5000-15 000/mm3), band form count≤ 1.5 x 109 cells/L (≤1500/mm3), ≤10 WBC per high power field on microscopic examination of spun urine sediment, and ≤5 WBC per high power field on microscopic examination of a stool smear (if diarrhea). The recommended evaluation included the culture of specimens of blood, cerebrospinal fluid, and urine for bacteria. Outcomes were determined. The negative predictive values of the low risk criteria for SBI and bacteremia were calculated. Results. Of 1057 eligible infants, 931 were well appearing, and, of these, 437 met the remaining low risk criteria. Five low risk infants had SBI including two infants with bacteremia. The negative predictive value of the low risk criteria was 98.9% (95% confidence interval, 97.2% to 99.6%) for SBI, and 99.5% (95% confidence interval, 98.2% to 99.9%) for bacteremia. Conclusions. These data confirm the ability of the low risk criteria to identify infants unlikely to have SBI. Infants who meet the low risk criteria can be carefully observed without administering antimicrobial agents.

A New Screening Approach For Fast And Accurate Prediction Of Positive And Negative Urine Cultures By SediMAX Compared With The Standard Urine Culture

2021 ◽  
Vol 1 (1) ◽  
pp. 46-55
Author(s):  
Massimo Pieri ◽  
Flaminia Tomassetti ◽  
Paola Cerini ◽  
Roberta Felicetti ◽  
Lucia Ceccaroni ◽  
...  
Keyword(s):  
Blood Cell ◽  
Bacterial Infections ◽  
Urine Culture ◽  
Screening Method ◽  
Significant Proportion ◽  
Bacterial Count ◽  
Urine Samples ◽  
Screening Methods ◽  
Urine Sediment ◽  
Tract Infections

Urinary tract infections (UTI) are the most frequent bacterial infections, and the detection of infection in urine samples is expensive and time-consuming. Also, in laboratories a significant proportion of samples processed yield negative results. For this, screening methods represent an important improvement towards the final UTI diagnosis. SediMAX is an automated microscopy, easier to use in laboratories due to its basic procedure and it is widely used for urine sediment analysis. In our study, we evaluated the performance of SediMAX, applying some screening parameters, compared with the gold standard methods, urine culture, to identify all the positive cases for UTI. We analysed 1185 urine samples from our daily laboratory routine. The basis of our screening model was to establish a cut-off for bacterial count (BACT), as 300 bacteria/µL in order to avoid missing positive cases. However, the sensitivity and the specificity achieved were not enough to identify all UTI infection in urine samples. So, in addition to BACT we have considered other parameters, such as White Blood Cell (WBC), Red Blood Cell (RBC), Yeasts (YEST), Age and Nitrates (NIT). The second screening method reached a sensitivity of 100%, that could be reliably employed in detect of UTIs.

scholarly journals Pattern of Esbls in Uro-Pathogens Obtained from a Nigerian Tertiary Hospital

2021 ◽  
Vol 16 (2) ◽  
pp. 139-147
Author(s):  
O. Olasehinde ◽  
A. Lamikanra
Keyword(s):  
Tertiary Hospital ◽  
Agarose Gel ◽  
Esbl Production ◽  
Gram Negative ◽  
Positive Urine ◽  
Key Factor ◽  
Agar Dilution ◽  
Tract Infections ◽  
Esbl Producers ◽  
Urine Specimens

Background: Multidrug resistance remains a challenge in the treatment of Urinary Tract Infections (UTI) in Nigeria, a key factor being the occurrence of ESBL producers. Earlier reports have emphasized the occurrence of major ESBLs, little is known about the minor subtypes' occurrence in this regard.Objective: This study sought to evaluate the occurrence of major and minor ESβL producers among a cohort of uropathogens collected from a Nigerian Teaching Hospital using molecular techniques.Material and Methods: Cultures from 1000 UTI positive urine specimens were collected from the hospital laboratory between May 2015 and December 2017. All samples were subjected to standard isolation culturing techniques and identified. They were further tested for susceptibility to 8 antibiotics. Of these, gram-negative isolates with presumptive ESBL production were evaluated for confirmatory ESBL production using Chromogenic ESBL agar (Oxoid) and Agar Dilution tests. One hundred and twenty-five of them were evaluated for the presence of six ESBL genes (TEM, SHV, CTX-M 15, PER, GES, VEB) using Multiplex PCR/Agarose Gel Electrophoresis.Results: The results revealed that 40 out of 97 (41%) detected ESBL genes were of the ESBL minor category (VEB, PER, and GES). These ESBL producers were also observed to be resistant to at least five of the 8 antibiotics tested.Conclusion: More attention should be paid to the emergence minor ESBL producers among uropathogens in this environment as they represent a potential underlying influence on the observed treatment failure in the treatment of UTI.   

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