Quantification of bacterial uropathogens in preclinical samples using real-time PCR assays.

Uropathogenic Escherichia coli (UPEC) and Staphylococcus saprophyticus (S. saprophyticus) are responsible for the majority of community-acquired urinary tract infections (UTI). Agar plating, a gold standard for detection of bacterial uropathogens, is labor intensive, limited for distinguishing between environmental contaminants and pathogens, and fails to effectively detect mixed infections. A reliable method for specific and sensitive quantitative assessment of infections would allow cost-effective evaluation of large numbers of experimental samples. A methodology such as quantitative PCR (qPCR) addresses the limitations of agar plating. We developed and validated highly specific and sensitive qPCR assays to assist researchers in the evaluation of potential vaccines and interventions in preclinical models of UPEC and S. saprophyticus UTI. The developed UPEC PCR targeted a highly conserved region of the UPEC hemolysin D (hlyD) gene that reproducibly detected type strains CFT073 and J96 over a 9 log range with high precision. To quantify S. saprophyticus genomes, a separate qPCR assay targeting the Trk transport gene was developed with an 8 log range. Neither assay detected bacterial species predicted to be sample contaminants. Using our optimized workflow that includes automated steps, up to 200 urine or tissue samples can be processed in as few as 3 h. Additionally, sequence comparisons of our primers and probe to other UTI bacterial strains indicated the broad applicability of these assays. These optimized qPCR assays provide a cost-effective and time-saving method for quantification of bacterial burdens in tissues and body fluids to assess the effectiveness of candidate vaccines or interventions.

Assessment of appropriate use of amylase and lipase testing in the diagnosis of acute pancreatitis at an academic teaching hospital.

OBJECTIVE: Despite evidence-based guidelines stating that lipase alone should be used in the diagnosis of suspected acute pancreatitis, health care providers continue to order amylase or amylase and lipase together. The purpose of this study was to assess the utilization of appropriate laboratory testing related to the diagnosis of acute pancreatitis. METHODS: The study used a retrospective cross-sectional design. The timeframe was from January 1, 2020, to December 31, 2020. A retrospective chart review was used to collect data for the following: patient-provider encounter notes, patient demographics, provider demographics, differential and final diagnosis, and laboratory test results. Data analysis include stratification of categorical variables and calculation of cost savings. RESULTS: For the 12-month period, this study found 2567 (9.3%) of all amylase and lipase tests to be unnecessary. Amylase tests (1881; 73.2%) made up the most unnecessary tests followed by lipase tests (686; 26.7%). An analysis of test-ordering behavior by providers revealed that 81.5% of all unnecessary tests were ordered by MDs. Finally, this study estimated a total cost savings of $128,350 if all unnecessary tests were eliminated. CONCLUSION: Our study demonstrated that amylase and lipase tests have been overutilized in the diagnosis of acute pancreatitis.

Blood usage and wastage at an academic teaching hospital before the initial wave of COVID-19 and during and after its quarantine periods.

BACKGROUND: Transfusion services aim to maintain sufficient blood inventory to support patients, even with challenges introduced by COVID-19. OBJECTIVES: To review blood usage and wastage before, during, and after COVID-19 surges, and to evaluate effects on inventory. METHODS: In a retrospective review, we evaluated the association between time periods corresponding to the initial wave of COVID-19 (pre-COVID-19, quarantine, and postquarantine) and blood usage/wastage. Data were stratified by period, and χ2 testing was used to examine the association between these time periods and blood usage/wastage. RESULTS: In the period before COVID-19, the transfusion service used more units, and in the period after quarantine, more units went to waste. Across all time periods, the most-used product was RBCs, and the most wasted product was plasma. A statistically significant association existed between usage (χ2 [6/3209 (0.2%)]) = 24.534; P ≤.001; Cramer V = 0.62), wastage (χ2 [6/775 (0.8%)]) = 21.673; P = .001; Cramer V = 0.118), and time period. The postquarantine period displayed the highest wastage costs ($51,032.35), compared with the pre-COVID-19 period ($29,734.45). CONCLUSION: Changes in blood inventory use and waste are significantly associated with the onset and continuation of COVID-19.

The Practicality of Preparing Skim Breast Milk at Home for Treatment of Infants Requiring Low Fat Diets.

Introduction: The benefits of breast milk (BM) for infants have long been established. However, for health-compromised infants with difficulty processing long-chain triglycerides, BM is often discontinued, and skimmed breast milk (SBM) is used as a dietary treatment. SBM is usually produced for inpatients in a hospital laboratory. The aim of this study was to determine the viability of skimming BM at home. Case Report: A female infant was diagnosed with congenital lipomatous asymmetric overgrowth, vascular malformations, epidermal nevi, and skeletal and spinal anomalies (CLOVES) syndrome, with symptoms of lymphatic malformation, chylothorax, and pleural effusion. The patient's family produced SBM at home after discharge; the SBM met the dietary treatment requirements and kept symptoms under control. Methods: A nonrefrigerated benchtop centrifuge was used to produce SBM at the patient's home. The optimal setting for the centrifuge was determined and then used to process BM samples from the infant's mother. The samples were randomly selected from each 10-day period over 6 months, and 18 samples were processed in total. The hospital laboratory processed the same samples of BM and analyzed the macronutrients with a comparison of the home-produced SBM to the hospital-produced SBM. Results: The home-produced SBM met the dietary treatment requirement of <1.0 g/dL of fat content. Fat was significantly lower, proteins were significantly higher, and carbohydrates and calories were not significantly different compared to hospital-produced SBM. Conclusions: It is viable to consistently produce SBM at home that meets the dietary treatment requirements of health-compromised infants.

Evaluation of Antinuclear Antibody and Subserology Reflex Testing for the Diagnosis of Systemic Autoimmune Rheumatic Disorders in an Academic Teaching Hospital.

OBJECTIVE: The aim of this study was to examine appropriate utilization of antinuclear antibody (ANA) screening tests with follow-up subserology tests (reflex testing) for diagnosing systemic autoimmune rheumatic disorder (SARD). METHODS: We conducted a retrospective chart review of 3003 SARD-test orders at an academic teaching hospital from January to December 2019. Testing patterns were categorized as American College of Rheumatology (ACR)-recommended reflex testing, panel testing, or single subserology testing. We described testing patterns, assessed their diagnostic accuracy, and explored factors associated with reflex testing. RESULTS: Reflex testing accounted for 79.7% of SARD test-ordering, whereas improper testing (panel or single subserology) accounted for the other 20.3%. Reflex testing was associated with significantly more SARD diagnoses than improper testing (P = .004). Testing patterns were significantly associated with race/ethnicity (P = .008), with reflex testing being less frequent than improper testing in Hispanics and Whites. CONCLUSION: In summary, one-fifth (20.3%) of testing patterns for suspected SARD did not follow the ACR-recommended guidelines for using reflex testing. Use of reflex testing was associated with an increased frequency of SARD diagnosis.