Search

We evaluated the DxU 850m Iris Urine Microscopy analyzer as a screening tool for excluding negative urine samples (n = 1337). At a cutoff of 10 3 colony counts·mL -1 , sensitivity was 55.1 %, specificity 68.6 %. The DxU 850m Iris does not offer acceptable prediction of culture-negative urine samples at the tested cutoff., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ulrik S Justesen reports equipment, drugs, or supplies was provided by Beckman Coulter Inc. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: The EFLM Task and Finish Group Urinalysis has updated the ECLM European Urinalysis Guidelines (2000) on urinalysis and urine bacterial culture, to improve accuracy of these examinations in European clinical laboratories, and to support diagnostic industry to develop new technologies., Recommendations: Graded recommendations were built in the following areas., Medical Needs and Test Requisition: Strategies of urine testing are described to patients with complicated or uncomplicated urinary tract infection (UTI), and high or low-risk to kidney disease., Specimen Collection: Patient preparation, and urine collection are supported with two quality indicators: contamination rate (cultures), and density of urine (chemistry, particles)., Chemistry: Measurements of both urine albumin and α1-microglobulin are recommended for sensitive detection of kidney disease in high-risk patients. Performance specifications are given for urine protein measurements and quality control of multiproperty strip tests., Particles: Procedures for microscopy are reviewed for diagnostic urine particles, including urine bacteria. Technologies in automated particle counting and visual microscopy are updated with advice how to verify new instruments with the reference microscopy., Bacteriology: Chromogenic agar is recommended as primary medium in urine cultures. Limits of significant growth are reviewed, with an optimised workflow for routine specimens, using leukocyturia to reduce less important antimicrobial susceptibility testing. Automation in bacteriology is encouraged to shorten turn-around times. Matrix assisted laser desorption ionization time-of-flight mass spectrometry is applicable for rapid identification of uropathogens. Aerococcus urinae, A. sanguinicola and Actinotignum schaalii are taken into the list of uropathogens. A reference examination procedure was developed for urine bacterial cultures., (© 2024 Walter de Gruyter GmbH, Berlin/Boston.)

Background: Azole resistance complicates treatment of patients with invasive aspergillosis with an increased mortality. Azole resistance in Aspergillus fumigatus is a growing problem and associated with human and environmental azole use. Denmark has a considerable and highly efficient agricultural sector. Following reports on environmental azole resistance in A. fumigatus from Danish patients, the ministry of health requested a prospective national surveillance of azole-resistant A. fumigatus and particularly that of environmental origin., Objectives: To present the data from the first 2 years of the surveillance programme., Methods: Unique isolates regarded as clinically relevant and any A. fumigatus isolated on a preferred weekday (background samples) were included. EUCAST susceptibility testing was performed and azole-resistant isolates underwent cyp51A gene sequencing., Results: The azole resistance prevalence was 6.1% (66/1083) at patient level. The TR 34 /L98H prevalence was 3.6% (39/1083) and included the variants TR 34 /L98H, TR 34 3 /L98H and TR 34 /L98H/S297T/F495I. Resistance caused by other Cyp51A variants accounted for 1.3% (14/1083) and included G54R, P216S, F219L, G54W, M220I, M220K, M220R, G432S, G448S and Y121F alterations. Non-Cyp51A-mediated resistance accounted for 1.2% (13/1083). Proportionally, TR 34 /L98H, other Cyp51A variants and non-Cyp51A-mediated resistance accounted for 59.1% (39/66), 21.2% (14/66) and 19.7% (13/66), respectively, of all resistance. Azole resistance was detected in all five regions in Denmark, and TR 34 /L98H specifically, in four of five regions during the surveillance period., Conclusion: The azole resistance prevalence does not lead to a change in the initial treatment of aspergillosis at this point, but causes concern and leads to therapeutic challenges in the affected patients., (© 2022 The Authors. Mycoses published by Wiley-VCH GmbH.)

As part of a national surveillance programme initiated in 2004, fungal blood isolates from 2016-2018 underwent species identification and EUCAST susceptibility testing. The epidemiology was described and compared to data from previous years. In 2016-2018, 1454 unique isolates were included. The fungaemia rate was 8.13/100,000 inhabitants compared to 8.64, 9.03, and 8.38 in 2004-2007, 2008-2011, and 2012-2015, respectively. Half of the cases (52.8%) involved patients 60-79 years old and the incidence was highest in males ≥70 years old. Candida albicans accounted for 42.1% of all isolates and Candida glabrata for 32.1%. C. albicans was more frequent in males ( p = 0.03) and C. glabrata in females ( p = 0.03). During the four periods, the proportion of C. albicans decreased ( p < 0.001), and C. glabrata increased ( p < 0.001). Consequently, fluconazole susceptibility gradually decreased from 68.5% to 59.0% ( p < 0.001). Acquired fluconazole resistance was found in 4.6% Candida isolates in 2016-2018. Acquired echinocandin resistance increased during the four periods 0.0%, 0.6%, 1.7% to 1.5% ( p < 0.0001). Sixteen echinocandin-resistant isolates from 2016-2018 harboured well-known FKS resistance-mutations and one echinocandin-resistant C. albicans had an FKS mutation outside the hotspot (P1354P/S) of unknown importance. In C. glabrata specifically, echinocandin resistance was detected in 12/460 (2.6%) in 2016-2018 whereas multidrug-class resistance was rare (1/460 isolates (0.2%)). Since the increase in incidence during 2004-2011, the incidence has stabilised. In contrast, the species distribution has changed gradually over the 15 years, with increased C. glabrata at the expense of C. albicans . The consequent decreased fluconazole susceptibility and the emergence of acquired echinocandin resistance complicates the management of fungaemia and calls for antifungal drug development.

Mucormycosis is a life threatening infection in patients with haematological disease. We introduced a Mucorales-PCR and an aggressive, multidisciplinary management approach for mucormycosis during 2016-2017 and evaluated patient outcomes in 13 patients diagnosed and treated in 2012-2019. Management principle: repeated surgical debridement until biopsies from the resection margins were clean as defined by negative Blankophor microscopy, Mucorales-PCR (both reported within 24 h), and cultures. Cultured isolates underwent EUCAST E.Def 9.3.1 susceptibility testing. Antifungal therapy (AFT) (mono/combination) combined with topical AFT (when possible) was given according to the minimal inhibitory concentration (MIC), severity of the infection, and for azoles, specifically, it was guided by therapeutic drug monitoring. The outcome was evaluated by case record review. All patients underwent surgery guided by diagnostic biopsies from tissue and resection margins (195 samples in total). Comparing 2012-2015 and 2016-2019, the median number of patients of surgical debridements was 3 and 2.5 and of diagnostic samples: microscopy/culture/PCR was 3/3/6 and 10.5/10/10.5, respectively. The sensitivity of microscopy (76%) and Mucorales-PCR (70%) were similar and microscopy was superior to that of culture (53%; p = 0.039). Initial systemic AFT was liposomal amphotericin B ( n = 12) or posaconazole ( n = 1) given as monotherapy ( n = 4) or in combination with isavuconazole/posaconazole ( n = 3/6) and terbinafine ( n = 3). Nine patients received topical amphotericin B. All received isavuconazole or posaconazole consolidation therapy ( n = 13). Mucormycosis related six month mortality was 3/5 in 2012-2015 and 0/7 patients in 2016-2019 (one patient was lost for follow-up). Implementation of combination therapy (systemic+topical AFT/combination systemic AFT) and aggressive surgical debridement guided by optimised diagnostic tests may improve the outcome of mucormycosis in haematologic patients.

Azole-resistant (azole-R) Aspergillus is an increasing challenge worldwide. Patients with cystic fibrosis (CF) are at risk of Aspergillus colonization and disease due to a favorable lung environment for microorganisms. We performed a nationwide study in 2018 of azole-non-susceptible Aspergillus in CF patients and compared with data from two prior studies. All airway samples with mold isolates from patients monitored at the two CF centers in Denmark (RH, Jan-Sept and AUH, Jan-Jun) were included. Classical species identification (morphology and thermo-tolerance) was performed and MALDI-TOF/β-tubulin sequencing was performed if needed. Susceptibility was determined using EUCAST E.Def 10.1, and E.Def 9.3.2. cyp51A sequencing and STR Af genotyping were performed for azole-non-susceptible isolates and relevant sequential isolates. In total, 340 mold isolates from 159 CF patients were obtained. The most frequent species were Aspergillus fumigatus (266/340, 78.2%) and Aspergillus terreus (26/340, 7.6%). Azole-R A. fumigatus was cultured from 7.3% (10/137) of patients, including 9.5% (9/95) of patients at RH and 2.4% at AUH (1/42), respectively. In a 10-year perspective, azole-non-susceptibility increased numerically among patients at RH (10.5% in 2018 vs 4.5% in 2007-2009). Cyp51A resistance mechanisms were found in nine azole-R A. fumigatus from eight CF patients. Five were of environmental origin (TR 34 /L98H), three were human medicine-driven (two M220K and one M220R), and one was novel (TR 34 3 /L98H) and found in a patient who also harbored a TR 34 /L98H isolate. STR Af genotyping identified 27 unique genotypes among 45 isolates and ≥2 genotypes in 8 of 12 patients. This included one patient carrying two unique TR 34 /L98H isolates, a rare phenomenon. Genotyping of sequential TR 34 3 /L98H and TR 34 /L98H isolates from the same patient showed only minor differences in 1/9 markers. Finally, azole-R A. terreus was found in three patients including two with Cyp51A alterations (M217I and G51A, respectively). Azole-R A. fumigatus is increasing among CF patients in Denmark with the environmentally associated resistance TR 34 /L98H mechanism being dominant. Mixed infections (wildtype/non-wildtype and several non-wildtypes) and a case of potential additional tandem repeat acquisition in vivo were found. However, similar genotypes were identified from another patient (and outside this study), potentially suggesting a predominant TR 34 /L98H clone in DK. These findings suggest an increasing prevalence and complexity of azole resistance in A. fumigatus ., (Copyright © 2020 Risum, Hare, Gertsen, Kristensen, Johansen, Helweg-Larsen, Abou-Chakra, Pressler, Skov, Jensen-Fangel and Arendrup.)

Objective: We have recently introduced intrapartum PCR-testing for group B streptococcus (GBS) in women in labor with prolonged rupture of membranes or preterm delivery to offer intrapartum antibiotic prophylaxis only for GBS positive women.The goal of the present study is to report our experience and results from the first half year of GBS testing., Study Design: This is a retrospective study. Rectovaginal swabs from 321 women presenting in the labor ward with pre-labor rupture of membranes for >14 h/rupture of membranes during delivery for >14 h, or labor between gestational weeks 35 0/7 and 36 6/7 from February 7, 2017 to August 6, 2017, were tested.We performed a molecular GBS test (Xpert GBS®, Cepheid Ltd., Sunnyvale, USA).Data from patient files including mode of delivery, use of antibiotics, infection of mother and child are presented in more detail.Data on the PCR results from the first year of testing were also collected., Results: In the first half-year of testing a positive GBS test result was found in 58 (18.1%) and a negative test result in 263 women (81.9%). No invalid test result was achieved.The indication for performing PCR testing was PROM > 14 h or rupture of membranes during labor for >14 h in 266 women (82.9%) and labor in gestational weeks 35 0/7-36 6/7 in 44 women (13.7%). In the remaining 11 women, the PCR test was performed for other reasons.Of the 321 women tested 126 (39%) received antibiotics during labor.Ten women (3.4%) were treated after delivery on suspicion of infection. 25 newborns (7.8%) were treated with antibiotics. In 11 cases, the treatment was stopped after 2-4 days as there were no signs of infection., Conclusions: The introduction of the intrapartum GBS test in selected groups of women who gave birth in our department has been well accepted by the women, the midwifes and doctors. The result of the test is available within two hours, and as we only offer intrapartum antibiotic prophylaxis to GBS-positive women, we have reduced the use of antibiotics to approximately 40% in the groups tested, without an increase of infection in mother or child., (© 2019 The Authors.)

We report a fatal aspergillosis case in which STRAf typing and whole-genome sequencing substantiated in vivo emergence of an azole-resistant Aspergillus fumigatus with a 120-bp tandem repeat in the promoter region of cyp51A. This event, previously restricted to the environment, challenges current understanding of azole resistance development in A. fumigatus.

New data from the years 2012 to 2015 from the Danish National Fungemia Surveillance are reported, and epidemiological trends are investigated in a 12-year perspective (2004 to 2015). During 2012 to 2015, 1,900 of 1,939 (98%) fungal bloodstream isolates were included. The average incidence was 8.4/100,000 inhabitants, and this appears to represent a stabilizing trend after the increase to 10.1/100,000 in 2011. The incidence was higher in males than females (10.0 versus 6.8) and in patients above 50 years, and those changes were mainly driven by an increasing incidence among 80-to-89-year-old males (65.3/100,000 in 2014 to 2015). The proportion of Candida albicans isolates decreased from 2004 to 2015 (64.4% to 42.4%) in parallel with a doubling of the proportion of Candida glabrata isolates (16.5% to 34.6%, P < 0.0001). C. glabrata was more common among females (34.0% versus 30.4% in males). Following an increase in 2004 to 2011, the annual drug use stabilized during the last 2 to 3 years of that time period but remained higher than in other Nordic countries. This was particularly true for the fluconazole and itraconazole use in the primary health care sector, which exceeded the combined national levels of use of these compounds in each of the other Nordic countries. Fluconazole susceptibility decreased (68.5%, 65.2%, and 60.6% in 2004 to 2007, 2008 to 2011, and 2012 to 2015, respectively, P < 0.0001), and echinocandin resistance emerged in Candida (0%, 0.6%, and 1.7%, respectively, P < 0.001). Amphotericin B susceptibility remained high (98.7%). Among 16 (2.7%) echinocandin-resistant C. glabrata isolates (2012 to 2015), 13 harbored FKS mutations and 5 (31%) were multidrug resistant. The epidemiological changes and the increased incidence of intrinsic and acquired resistance emphasize the importance of continued surveillance and of strengthened focus on antifungal stewardship., (Copyright © 2018 American Society for Microbiology.)

Introduction: Many pregnant women are treated with antibiotics during labor to prevent transmission of group B streptococcus (GBS, Streptococcus agalactiae) to their baby during passage of the birth canal, and so reduce the risk of serious infection of the newborn. Methods for intrapartum testing for GBS have been introduced to select women to whom intrapartum antibiotic prophylaxis should be offered. For such an intrapartum test to be useful in clinical practice, it has to be specific as well as sensitive. The aim of the present study is to evaluate the accuracy of the polymerase chain reaction (PCR) assay compared with an optimized culture method for GBS., Material and Methods: In the period from 12 May 2015 to 18 December 2015 we collected rectovaginal swabs from 106 women in the labor ward presenting in labor between gestational week 35 +0 and 36 +6 or presenting with prelabor/preterm prelabor rupture of membranes (PROM/PPROM) for > 14 h after gestational week 34 +0 . We performed GBS culture (reference standard) and a molecular GBS test (Xpert GBS, Cepheid Ltd., Sunnyvale, CA, USA)., Results: Based on intrapartum culture, 23.6% (25/106) were colonized with GBS. Intrapartum PCR showed a colonization rate of 25.7% (27/105). The sensitivity of the test was 100% (86.28-100%). The specificity of the test was 97.5% (91.26-99.70%). The positive predictive value was 92.6%. In one case, we had no result with PCR testing, giving an invalid test rate of < 1%., Conclusion: The PCR test has sufficient accuracy to direct intrapartum antibiotic prophylaxis for GBS transmission during delivery., (© 2017 Nordic Federation of Societies of Obstetrics and Gynecology.)

We report two cases of cerebral abscesses with polymicrobial aetiology including Mycoplasma salivarium. In both cases, Mycoplasma was found incidentally, suggesting that a broader aetiological spectrum could be found in brain abscesses by use of molecular techniques targeting fastidious pathogens., (© 2011 The Authors. Clinical Microbiology and Infection © 2011 European Society of Clinical Microbiology and Infectious Diseases.)