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OBJECTIVES: We propose fast and accurate molecular detection of the Y132F ERG11p substitution directly on pure-cultured Candida parapsilosis isolates. We also assessed a discriminative genotyping scheme to track circulating genotypes. METHODS: A total of 223 C. parapsilosis isolates (one patient each) from 20 hospitals, located in Spain and Italy were selected. Isolates were fluconazole-resistant (n = 94; harbouring the Y132F ERG11p substitution [n = 85], the G458S substitution [n = 6], the R398I substitution [n = 2], or the wild-type ERG11 gene sequence) or fluconazole-susceptible (n = 129). Two targeted-A395T-mutation PCR formats (conventional and real-time) were engineered and optimized on fluconazole-susceptible and fluconazole-resistant pure-cultured isolates, thus skipping DNA extraction. Two genotyping schemes were compared: Scheme 1 (CP1, CP4a, CP6, and B markers), and Scheme 2 (6A, 6B, 6C, CP1, CP4a, and CP6 markers). RESULTS: The screening performed using both PCR formats showed 100% specificity (fluconazole-susceptible isolates; n = 129/129) and sensitivity (Y132F isolates; n = 85/85) values; however, results were available in 3 and 1.5 hours with the conventional and real-time PCR formats, respectively. Overall, Scheme 1 showed higher genetic diversity than Scheme 2, as shown by the number of alleles detected (n = 98; mean 23, range 13-38), the significantly higher observed and expected heterozygosity, and the probability of identity index (2.5 × 10-6). Scheme 2 markers did not provide further genotypic discrimination of Y132F fluconazole-resistant genotypes. CONCLUSION: Both proposed PCR formats allow us to speed up the accurate detection of substitution Y132F ERG11p in C. parapsilosis isolates with 100% specificity and sensitivity. In addition, we recommend CP1, CP4a, CP6, and B microsatellite markers for genotyping fluconazole-resistant isolates.
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BACKGROUND: Fluconazole-resistant Candida parapsilosis is a matter of concern. OBJECTIVES: To describe fluconazole-resistant C. parapsilosis genotypes circulating across hospitals in Spain and Rome and to study their azole-resistance profile associated with ERG11p substitutions. PATIENTS/METHODS: We selected fluconazole-resistant C. parapsilosis isolates (n = 528 from 2019 to 2023; MIC ≥8 mg/L according to EUCAST) from patients admitted to 13 hospitals located in five Spanish cities and Rome. Additionally, we tested voriconazole, posaconazole, isavuconazole, amphotericin B, micafungin, anidulafungin and ibrexafungerp susceptibility. RESULTS: Of the 53 genotypes found, 49 harboured the Y132F substitution, five of which were dominating city-specific genotypes involving almost half the isolates. Another genotype involved isolates harbouring the G458S substitution. Finally, we found two genotypes with the wild-type ERG11 gene sequence and one with the R398I substitution. All isolates were fully susceptible/wild-type to amphotericin B, anidulafungin, micafungin and ibrexafungerp. The azole-resistance patterns found were: voriconazole-resistant (74.1%) or voriconazole-intermediate (25.2%), posaconazole-resistant (10%) and isavuconazole non-wild-type (47.5%). Fluconazole-resistant and voriconazole non-wild-type isolates were likely to harbour substitution Y132F if posaconazole was wild type; however, if posaconazole was non-wild type, substitution G458S was indicated if isavuconazole MIC was >0.125 mg/L or substitution Y132F if isavuconazole MIC was ≤0.125 mg/L. CONCLUSIONS: We detected a recent clonal spread of fluconazole-resistant C. parapsilosis across some cities in Spain, mostly driven by dominating city-specific genotypes, which involved a large number of isolates harbouring the Y132F ERG11p substitution. Isolates harbouring substitution Y132F can be suspected because they are non-susceptible to voriconazole and rarely posaconazole-resistant.
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Azóis , Fluconazol , Glicosídeos , Nitrilas , Piridinas , Triazóis , Triterpenos , Humanos , Azóis/farmacologia , Fluconazol/farmacologia , Candida parapsilosis/genética , Cidades , Voriconazol/farmacologia , Anfotericina B , Anidulafungina , Micafungina , Itália , Hospitais , GenótipoRESUMO
More than one million sexually transmitted infections (STIs) occur every day, and Trichomonas vaginalis is responsible for more than 156 million cases each year worldwide. Nevertheless, epidemiological studies of this parasite in Europe are scarce. The aim of this study was to evaluate the impact that the COVID-19 pandemic may have had in the diagnosis and epidemiology of trichomoniasis. All available data from January 2018 to December 2021 for T. vaginalis isolation on gynecologic patients attending a Spanish Tertiary Hospital were analyzed. Pre-pandemic results (2018-2019) were compared to pandemic results (2020-2021). The pre-pandemic T. vaginalis prevalence in women was 1.15% (95% Confidence Interval, CI: 0.94-1.41), and significantly decreased in 2020-2021 (0.77%, 95% CI: 0.57-1.03; p = 0.025). Demographic nor clinical characteristics of women diagnosed with trichomoniasis did not statistically differ between the periods, although an increase in chlamydia co-infected patients was observed in the latest (from 8% in 2018-2019 to 19% in 2020-2021). This study has detected a decrease in the diagnosis of trichomoniasis; however, this is probably due to the increase in the healthcare pressure triggered by the pandemic. More than 75% of the cases diagnosed in 2021 occurred in the second half, which suggests that special attention should be given to the evolution in the coming years once normality has been restored in hospitals. Moreover, these results warn of the lack of routine diagnosis of trichomoniasis during pregnancy and the absence of specific protocols for possible co-infections, which could become a strategy to reduce the growing trend of STIs, including T. vaginalis detection, as an interesting marker of sexual risk behaviors.
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We previously conducted a multicenter surveillance study on Candida epidemiology and antifungal resistance in Madrid (CANDIMAD study; 2019-2021), detecting an increase in fluconazole-resistant Candida parapsilosis. We here present data on isolates collected in 2022. Furthermore, we report the epidemiology and antifungal resistance trends during the entire period, including an analysis per ward of admission. Candida spp. incident isolates from blood cultures and intra-abdominal samples from patients cared for at 16 hospitals in Madrid, Spain, were tested with the EUCAST E.Def 7.3.2 method against amphotericin B, azoles, micafungin, anidulafungin, and ibrexafungerp and were molecularly characterized. In 2022, we collected 766 Candida sp. isolates (686 patients; blood cultures, 48.8%). Candida albicans was the most common species found, and Candida auris was undetected. No resistance to amphotericin B was found. Overall, resistance to echinocandins was low (0.7%), whereas fluconazole resistance was 12.0%, being higher in blood cultures (16.0%) mainly due to fluconazole-resistant C. parapsilosis clones harboring the Y132F-R398I ERG11p substitutions. Ibrexafungerp showed in vitro activity against the isolates tested. Whereas C. albicans was the dominant species in most hospital wards, we observed increasing C. parapsilosis proportions in blood. During the entire period, echinocandin resistance rates remained steadily low, while fluconazole resistance increased in blood from 6.8% (2019) to 16% (2022), mainly due to fluconazole-resistant C. parapsilosis (2.6% in 2019 to 36.6% in 2022). Up to 7 out of 16 hospitals were affected by fluconazole-resistant C. parapsilosis. In conclusion, rampant clonal spreading of C. parapsilosis fluconazole-resistant genotypes is taking place in Madrid.
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Candida , Fluconazol , Humanos , Fluconazol/farmacologia , Antifúngicos/farmacologia , Anfotericina B/farmacologia , Candida parapsilosis/genética , Tração , Equinocandinas , Candida albicans/genética , Farmacorresistência Fúngica/genética , Testes de Sensibilidade MicrobianaRESUMO
OBJECTIVES: Antifungal susceptibility testing is mostly conducted on blood-cultured Candida spp isolates. Because the intra-abdominal cavity has been highlighted as a hidden echinocandin-resistant C. glabrata reservoir, we assessed whether testing sequential isolates from a given patient might increase the chances of detecting antifungal resistance. METHODS: Intra-abdominal initial and sequential isolates from the same species from patients included in the CANDIdaemia in MADrid study (January 2019 to June 2022) were studied. We assessed antifungal susceptibility to amphotericin B, azoles, anidulafungin, micafungin, and ibrexafungerp using European Committee on Antimicrobial Susceptibility Testing (EUCAST) methodology and molecularly characterized resistant isolates. RESULTS: We collected 308 isolates (C. albicans [n = 179/308; 58.1%], C. glabrata [n = 101/308; 32.8%], C. tropicalis [n = 17/308; 5.5%], and C. parapsilosis [n = 11/308; 3.6%]) from 112 patients distributed as incident (n = 125/308) and sequential (n = 183/308). Per patient resistance rates of fluconazole (13.4% [15/112] vs. 8% [9/112]); 5.4% proportions difference (95% CI, -2.7% to 13.5%, p 0.09) and echinocandins (8.9% [10/112] vs. 1.8% [2/112]); 7.1% proportions difference (95% CI; 1.2-12.9%; p 0.01) were higher when considering all available isolates than only incident isolates. Resistance was detected in 18 of 112 patients and would have been overlooked in 11 of 18 (61.1%) patients if only incident isolates had been studied. Of the patients who harboured fluconazole or echinocandin-resistant isolates, 14 of 15 and 8 of 10 had received or were receiving fluconazole or echinocandins, respectively. DISCUSSION: Testing sequential Candida isolates from intra-abdominal samples is required to detect antifungal resistance, particularly to echinocandins, in patients whose incident isolates turned out to be susceptible. Furthermore, patients with echinocandin-resistant infections had frequently used echinocandins and had common secondary resistance acquisition.
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Antifúngicos , Candida , Humanos , Antifúngicos/farmacologia , Fluconazol , Equinocandinas/farmacologia , Anfotericina B , Candida albicans , Candida parapsilosis , Candida tropicalis , Candida glabrata , Testes de Sensibilidade Microbiana , Farmacorresistência FúngicaRESUMO
Background: Candida parapsilosis is a frequent cause of candidemia worldwide. Its incidence is associated with the use of medical implants, such as central venous catheters or parenteral nutrition. This species has reduced susceptibility to echinocandins, and it is susceptible to polyenes and azoles. Multiple outbreaks caused by fluconazole-nonsusceptible strains have been reported recently. A similar trend has been observed among the C. parapsilosis isolates received in the last 2 years at the Spanish Mycology Reference Laboratory. Methods: Yeast were identified by molecular biology, and antifungal susceptibility testing was performed using the European Committee on Antimicrobial Susceptibility Testing protocol. The ERG11 gene was sequenced to identify resistance mechanisms, and strain typing was carried out by microsatellite analysis. Results: We examined the susceptibility profile of 1315 C. parapsilosis isolates available at our reference laboratory between 2000 and 2021, noticing an increase in the number of isolates with acquired resistance to fluconazole, and voriconazole has increased in at least 8 different Spanish hospitals in 2020-2021. From 121 recorded clones, 3 were identified as the most prevalent in Spain (clone 10 in Catalonia and clone 96 in Castilla-Leon and Madrid, whereas clone 67 was found in 2 geographically unrelated regions, Cantabria and the Balearic Islands). Conclusions: Our data suggest that concurrently with the coronavirus disease 2019 pandemic, a selection of fluconazole-resistant C. parapsilosis isolates has occurred in Spain, and the expansion of specific clones has been noted across centers. Further research is needed to determine the factors that underlie the successful expansion of these clones and their potential genetic relatedness.
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OBJECTIVES: We prospectively monitored the epidemiology and antifungal susceptibility of Candida spp. from blood cultures and intra-abdominal samples in patients admitted to hospitals in the Madrid area. METHODS: Between 2019 and 2021, we prospectively collected incident isolates [one per species, patient and compartment (blood cultures versus intra-abdominal samples)] from patients admitted to any of 16 hospitals located in Madrid. We studied the antifungal susceptibilities to amphotericin B, triazoles, micafungin, anidulafungin and ibrexafungerp following the EUCAST E.Def 7.3.2 procedure. RESULTS: A total of 2107 Candida spp. isolates (1895 patients) from blood cultures (51.7%) and intra-abdominal samples were collected. Candida albicans, the Candida glabrata complex, the Candida parapsilosis complex, Candida tropicalis and Candida krusei accounted for 96.9% of the isolates; in contrast, Candida auris was undetected. Fluconazole resistance in Candida spp. was higher in blood cultures than in intra-abdominal samples (9.1% versus 8.2%; Pâ>â0.05), especially for the C. parapsilosis complex (16.6% versus 3.6%, Pâ<â0.05), whereas echinocandin resistance tended to be lower in blood cultures (0.5% versus 1.0%; Pâ>â0.05). Resistance rates have risen, particularly for fluconazole in blood culture isolates, which increased sharply in 2021. Ibrexafungerp showed in vitro activity against most isolates. Species distributions and resistance rates varied among hospitals. CONCLUSIONS: Whereas no C. auris isolates were detected, fluconazole-resistant C. parapsilosis isolates have been spreading across the region and this has pulled up the rate of fluconazole resistance. In contrast, the rate of echinocandin resistance continues to be low.
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Candida parapsilosis , Equinocandinas , Humanos , Equinocandinas/farmacologia , Fluconazol , Candida , Antifúngicos/farmacologia , Candida auris , Testes de Sensibilidade Microbiana , Farmacorresistência FúngicaRESUMO
We have been monitoring the antifungal resistance in Candida parapsilosis isolates collected from inpatients at Madrid metropolitan area hospitals for the last 3 years. The study aimed to elucidate the presence of fluconazole-resistant C. parapsilosis genotypes in Madrid. From January 2019 to December 2021, a total of 354 C. parapsilosis isolates (n = 346 patients) from blood (76.6%) or intraabdominal samples were collected and genotyped using species-specific microsatellite markers. Antifungal susceptibilities to amphotericin B, the triazoles, micafungin, anidulafungin, and ibrexafungerp were performed according to EUCAST E.Def 7.3.2; the ERG11 gene was sequenced in fluconazole-resistant isolates. A total of 13.6% (n = 48/354) isolates (one per patient) were found to be resistant to fluconazole and non-wild-type to voriconazole but fully susceptible to ibrexafungerp. Resistant isolates were mostly sourced from blood (n = 45/48, 93.8%) and were detected in five hospitals. Two hospitals accounted for a high proportion of resistant isolates (n = 41/48). Resistant isolates harbored either the Y132F ERG11p amino acid substitution (n = 43) or the G458S substitution (n = 5). Isolates harboring the Y132F substitution clustered into a clonal complex involving three genotypes (one genotype accounted for n = 39/43 isolates) that were found in four hospitals. Isolates harboring the G458S substitution clustered into another genotype found in a fifth hospital. C. parapsilosis genotypes demonstrating resistance to fluconazole have been spreading across hospitals in Madrid, Spain. Over the last 3 years, the frequency of isolation of such isolates and the number of hospitals affected is on the rise.
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Candida parapsilosis , Fluconazol , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Candida parapsilosis/genética , Farmacorresistência Fúngica/genética , Fluconazol/farmacologia , Genótipo , Hospitais , Humanos , Testes de Sensibilidade Microbiana , Espanha/epidemiologiaRESUMO
Severely ill COVID-19 patients are at high risk of nosocomial infections. The aim of the study was to describe the characteristics of candidemia during the pre-pandemic period (January 2019−February 2020) compared to the pandemic period (March 2020−September 2021). Antifungal susceptibilities were assessed using the EUCAST E.Def 7.3.2 broth dilution method. Fluconazole-resistant C. parapsilosis isolates (FRCP) were studied for sequencing of the ERG11 gene. The incidence of candidemia and C. parapsilosis bloodstream infection increased significantly in the pandemic period (p = 0.021). ICU admission, mechanical ventilation, parenteral nutrition and corticosteroids administration were more frequent in patients with candidemia who had been admitted due to COVID-19. Fifteen cases of FRCP fungemia were detected. The first case was recorded 10 months before the pandemic in a patient transferred from another hospital. The incidence of FRCP in patients admitted for COVID-19 was 1.34 and 0.16 in all other patients (p < 0.001). ICU admission, previous Candida spp. colonization, arterial catheter use, parenteral nutrition and renal function replacement therapy were more frequent in patients with candidemia due to FRCP. All FRCP isolates showed the Y132F mutation. In conclusion, the incidence of candidemia experienced an increase during the COVID-19 pandemic and FRCP fungemia was more frequent in patients admitted due to COVID-19.
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OBJECTIVES: Vaginal and endocervical infections are considered a global health problem, especially after recent evidence of their association with preterm delivery and other adverse obstetric outcomes. Still, there is no consensus on the efficacy of a screening strategy for these infections in the first trimester of pregnancy. This study evaluated their prevalence and whether screening and treatment resulted as effective in reducing pregnancy and perinatal complications. METHODS: A single-center prospective observational study was designed; a sample size of 400 first-trimester pregnant women was established and they were recruited between March 2016-October 2019 at the Puerta de Hierro University Hospital (Spain). They were screened for vaginal and endocervical infections and treated in case of abnormal flora. Pregnancy and delivery outcomes were compared between abnormal and normal flora groups by univariate analysis. RESULTS: 109 patients had an abnormal flora result (27.2%). The most frequently detected infection was Ureaplasma urealyticum (12.3%), followed by Candida spp. (11.8%), bacterial vaginosis (5%), Mycoplasma hominis (1.2%) and Trichomonas vaginalis (0.8%). Patients with abnormal flora had a 5-fold increased risk of preterm premature rupture of membranes (5.3% vs. 1.1% of patients with normal flora, Odds Ratio 5.11, 95% Confidence Interval 1.20-21.71, p = 0.028). No significant differences were observed regarding preterm delivery or neonatal morbidity. CONCLUSIONS: Considering the morbimortality related to prematurity and that the results of our study suggest that the early treatment of abnormal flora could improve perinatal outcomes, the implementation of a screening program during the first trimester should be considered.
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BACKGROUND: . Viral infections are recognized as the most common cause of acute gastroenteritis (AGE). Virus detection by immune analytical methods is recommended for diagnosis because of its simplicity and low cost. OBJECTIVES: . Two commercial immunochromatographic (ICG) techniques (Materlab) for rapid detection of rotavirus/adenovirus and norovirus respectively, were evaluated by comparison to the results obtained using PCR methods. In addition, clinical and epidemiologic characteristics of AGE infections have been described. STUDY DESIGN: . A total of 100 faecal samples collected from patients with AGE (84% children) admitted into a Spanish Hospital between February and July 2018, were studied for rotavirus-A, adenovirus and norovirus GI/GII by the ICG tests as well as by PCR and sequencing. Other enteric viruses (enterovirus and astrovirus) were investigated by PCR methods. Gastrointestinal bacteria and parasites were also tested. RESULTS: . Evaluated ICG tests yielded high specificity (>97%). Sensitivity values were high for rotavirus/adenovirus (>80%) but lower for norovirus (57%). Overall, and taking into account coinfections, viruses (32%), bacteria (14%) and parasites (1%) could be detected. Rotavirus-A were the most frequently identified viruses (16%), followed by enterovirus (12%), norovirus (4%), adenovirus 41 (4%) and astrovirus (1%). In five vaccinated children, a rotavirus was detected. CONCLUSIONS: . ICG technique is a useful tool for the routine diagnosis of AGE infections at hospital, but for surveillance and epidemiological studies, it is needed the use of amplification and sequencing methods, which also allow monitoring of new strains or variants emergence. In this study, an etiological pathogen was determined only in 44% of samples.
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Cromatografia de Afinidade/métodos , Fezes/virologia , Gastroenterite/diagnóstico , Gastroenterite/virologia , Viroses/diagnóstico , Adenoviridae/genética , Infecções por Adenovirus Humanos/diagnóstico , Adolescente , Infecções por Caliciviridae/diagnóstico , Criança , Pré-Escolar , Infecções por Enterovirus/diagnóstico , Hospitalização , Humanos , Lactente , Norovirus/genética , Kit de Reagentes para Diagnóstico , Rotavirus/genética , Sensibilidade e EspecificidadeRESUMO
Trichomoniasis is the most prevalent curable sexually transmitted infection (STI) worldwide and a risk factor for the acquisition of other STIs and adverse pregnancy outcomes. The objectives of this study were to determine the prevalence of T. vaginalis and related coinfections in women attending a third-level hospital of Madrid (Spain). A retrospective study of 24,173 vaginal exudates from women with suspected vaginitis was conducted between 2013 and 2017. Likewise, among T. vaginalis positive samples, co-occurrence with gonorrhea, chlamydia, syphilis, VIH, Mycoplasma hominis, and Ureaplasma urealyticum was checked. Moreover, seven T. vaginalis isolates from 2017 were randomly collected for endobionts, drug resistance, and microsatellite (MS) instability determinations. The prevalence of T. vaginalis was 0.8% between 2013 and 2017. Less than 20% of patients with trichomoniasis were submitted to a complete screening for other genital pathogens. From that, two patients were coinfected with chlamydia and three with syphilis. Surprisingly, 6.4% of positive samples were diagnosed among pregnant women, showing an alarming increase from 3.2% (2014) to 10% (2017). Among the isolates randomly analyzed, five carried T. vaginalis virus, five harbored mycoplasmas, and one was metronidazole-resistant. The molecular genotyping showed a high variability in the three MS evaluated. To our knowledge, this is the first study in Spain that evaluates the prevalence of trichomoniasis in general and pregnant population and includes biomolecular determinations. These results warn about the increasing prevalence and highlight the importance of including T. vaginalis detection in routine gynecological revisions with special emphasis on childbearing age women and patients with previous STIs.
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Metronidazol/farmacologia , Simbiose , Centros de Atenção Terciária , Tricomoníase/epidemiologia , Trichomonas vaginalis , Adulto , Coinfecção , Resistência a Medicamentos , Feminino , Gonorreia/complicações , Humanos , Pessoa de Meia-Idade , Mycoplasma hominis/isolamento & purificação , Gravidez , Complicações Parasitárias na Gravidez/epidemiologia , Taxa de Gravidez , Prevalência , Estudos Retrospectivos , Espanha/epidemiologia , Tricomoníase/complicações , Tricomoníase/tratamento farmacológicoRESUMO
Rapid diagnosis is one of the best ways to improve patient management and prognosis as well as to combat the development of bacterial resistance. The aim of this study was to study parameters that impact the achievement of reliable identification using a combination of flow cytometry and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-ToF-MS).The study was carried out in nine hospitals in Spain and included 1,050 urine samples with bacterial counts of 5x106 bacteria/ml. MALDI-ToF-MS-based identification was performed according to a previously described protocol. Valid identification by direct MALDI-ToF-MS was obtained in 72.8% of samples, in 80.3% of samples found to be positive by culture, 32.2% of contaminated samples, and 19.7% of negative samples. Among the positives samples with a valid identification the concordance at the species level was 97.2%. The parameters related to success of direct identification were: high bacterial count, the presence of Escherichia coli as a pathogen and rod-bacteria morphology provided by flow cytometry. The parameters related to failure were a high epithelial cell (EC) count, a high white blood cell (WBC) count and urine samples obtained from in-patients. In summary, this multicentre study confirms previously published data on the usefulness and accuracy of direct MALDI-ToF-MS-based identification of bacteria from urine samples. It seems important to evaluate not only the bacterial count, but also other parameters, such as EC and WBC counts, bacterial species and morphology, and the health care setting, to decide whether the sample is suitable for direct identification.
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Bacteriúria/diagnóstico , Bacteriúria/microbiologia , Infecções Urinárias/diagnóstico , Infecções Urinárias/microbiologia , Urina/microbiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Técnicas Bacteriológicas , Infecções por Escherichia coli/diagnóstico , Infecções por Escherichia coli/microbiologia , Feminino , Citometria de Fluxo , Humanos , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Espanha , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizAssuntos
Aerococcus/isolamento & purificação , Infecções por Bactérias Gram-Positivas/microbiologia , Nefrectomia , Complicações Pós-Operatórias/microbiologia , Infecções Urinárias/microbiologia , Idoso , Carcinoma de Células Renais/cirurgia , Diagnóstico Diferencial , Gastrectomia , Infecções por Bactérias Gram-Positivas/diagnóstico , Hepatite Viral Humana/complicações , Humanos , Hospedeiro Imunocomprometido , Neoplasias Renais/cirurgia , Masculino , Infecções Estreptocócicas/diagnóstico , Estreptococos ViridansRESUMO
OBJECTIVE: To evaluate the Sysmex UF-1000i system as a urine screening method for the diagnosis of urinary tract infection, and epithelial cells as a predictive value of contamination in woman of childbearing age. METHODS: A total of 1730 urine samples were processed using a urine culture as a reference. RESULTS: With 50 bacteria/µl as a cut-off point, the results were: sensitivity 91.3%, specificity 73.1%, negative predictive value 96.2%. For a specificity of 90% for epithelial cells, the results were: sensitivity 31.0%, positive predictive value 67.0%, negative predictive value 66.0%. CONCLUSION: The evaluated system is fast and effective. Epithelial cells could be used to predict contamination.
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Infecções Urinárias/diagnóstico , Infecções Urinárias/urina , Urina/citologia , Adolescente , Adulto , Automação Laboratorial/instrumentação , Bactérias/isolamento & purificação , Técnicas Bacteriológicas/instrumentação , Células Epiteliais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Urinálise , Urina/microbiologia , Adulto JovemRESUMO
We study the epidemiology, molecular basis, clinical risk factors, and outcome involved in the clonal dissemination of VIM-1-producing Klebsiella pneumoniae isolates in the hospital setting. All patients infected/colonized by carbapenem-nonsusceptible K. pneumoniae (CNSKP) in 2009 were included. Molecular epidemiology was studied by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Antibiotic resistance genes were analyzed by PCR and sequencing. Plasmids were studied by PFGE with S1 nuclease digestion and for incompatibility group by a PCR-based replicon typing scheme. Risk factors associated with CNSKP colonization/infection were assessed by an observational case-control study. All 55 patients studied were infected (n = 28) or colonized (n = 27) by VIM-1-producing K. pneumoniae. All but one acquired isolates of a single clone (PFGE cluster 1 [C1], sequence type 15 [ST15]), while another clone (PFGE C2, ST340) was detected in four patients. C1 isolates also produced the new extended-spectrum ß-lactamase SHV-134. bla(VIM-1) was carried in a class 1 integron and an untypeable plasmid of â¼50 bp. The number of days that the patient received mechanical ventilation, the use of parenteral nutrition, previous treatment with linezolid, and treatment with extended-spectrum cephalosporins for more than 7 days were detected to be independent risk factors for CNSKP acquisition. The VIM-1-producing K. pneumoniae ST15 clone has a high capacity to spread among intensive care unit patients with severe underlying conditions. A high rate of associated mortality and great difficulty in controlling the spread of this clone, without permanent behavioral changes in the personnel, were observed.