Bacteriuria and phenotypic antimicrobial susceptibility testing in 45 min by point-of-care Sysmex PA-100 System: first clinical evaluation.

PURPOSE: This study compared the results of the new Sysmex PA-100 AST System, a point-of-care analyser, with routine microbiology for the detection of urinary tract infections (UTI) and performance of antimicrobial susceptibility tests (AST) directly from urine. METHODS: Native urine samples from 278 female patients with suspected uncomplicated UTI were tested in the Sysmex PA-100 and with reference methods of routine microbiology: urine culture for bacteriuria and disc diffusion for AST. RESULTS: The analyser delivered bacteriuria results in 15 min and AST results within 45 min. Sensitivity and specificity for detection of microbiologically confirmed bacteriuria were 84.0% (89/106; 95% CI: 75.6-90.4%) and 99.4% (155/156; 95% CI: 96.5-100%), respectively, for bacterial species within the analyser specifications. These are Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Enterococcus faecalis and Staphylococcus saprophyticus, which are common species causing uncomplicated UTI. Overall categorical agreement (OCA) for AST results for the five antimicrobials tested in the Sysmex PA-100 (amoxicillin/clavulanic acid, ciprofloxacin, fosfomycin, nitrofurantoin and trimethoprim) ranged from 85.4% (70/82; 95%CI: 75.9-92.2%) for ciprofloxacin to 96.4% (81/84; 95% CI: 89.9-99.3%) for trimethoprim. The Sysmex PA-100 provided an optimal treatment recommendation in 218/278 cases (78.4%), against 162/278 (58.3%) of clinical decisions. CONCLUSION: This first clinical evaluation of the Sysmex PA-100 in a near-patient setting demonstrated that the analyser delivers phenotypic AST results within 45 min, which could enable rapid initiation of the correct targeted treatment with no further adjustment needed. The Sysmex PA-100 has the potential to significantly reduce ineffective or unnecessary antibiotic prescription in patients with UTI symptoms.

Improving the diagnosis of urinary tract infections by the use of enriched media and a 48-hour incubation period.

Introduction. The absence of a gold-standard methodology for the microbiological diagnosis of urinary tract infections (UTI) has led to insufficient standardization of criteria for the interpretation of results and processing methods, particularly incubation time and culture media.Hypothesis. 48-hour incubation time period and use of blood agar enhances the sensitivity of microorganisms isolated significantly.Aim. To determine the sensitivity of blood agar and Brilliance UTI chromogenic agar, incubating for different periods (24-48 hours), for the detection of positive urine cultures.Methodoloy. Comparisons were made between all possible combinations of media and incubation times. As the gold-standard reference, we used the routine methodology of our laboratory, which involves prior screening with available clinical data, flow cytometry, sediment analysis and/or Gram staining. Screened samples were then cultured on blood agar and chromogenic agar and incubated for 48 hours. Also, based on the results of Gram staining, additional media were added in selected cases.Results. The most significant difference was found between chromogenic agar incubated for 24 hours and blood agar incubated for 48 hours, with the latter method allowing the recovery of 10.14 % more microorganisms (P < 0.0001). Furthermore, the value of performing Gram staining to guide processing was demonstrated, as it avoided the loss of at least 5.14 % of isolates.Conclusions. At least in urological and nephrological patients it is essential to include enriched culture media (blood agar) or to extend the incubation times due to the improvement of the diagnostic sensitivity of urine cultures. Gram staining also can help detect the presence of fastidious microorganisms or mixed infections, indicating whether rich and/or selective media should be included to enhance the diagnostic sensitivity of cultures. If this methodology is not followed, it should be noted that besides fastidious species, fastidious strains of Escherichia coli, Proteus mirabilis, Pseudomonas aerugniosa and Stenotrophomonas maltophilia will also be missed.

[Neisseria meningitidis and the increase of oral sex. A case report]. / Neisseria meningitidis y el aumento del sexo oral. A propósito de un caso.

Urethritis is an entity characterized by dysuria and purulent urethral discharge, generally acquired sexually. Neisseria gonorrhoeae is one of the most frequently responsible microorganisms. Neisseria meningitidis is a gram-negative diplococcus usually isolated in the pharynx, that occasionally causes meningococcal meningitis, being unusual it's isolation in the anogenital area where it could be a genitourinary pathogen. We present the case of a 25-years-old heterosexual male who, after a heterosexual intercourse with an occasional non-professional partner, including oral and vaginal sex, presented with symptoms of urethritis, orienting to a sexually transmitted infection. The bacteriological culture for N. gonorrhoeae was negative and the PCR for Chlamydia trachomatis was positive. Subsequently, the lab reported a positive bacteriological culture for sero-group C N. meningitidis, sensitive to ceftriaxone and a negative PCR for N. gonorrhoeae. N. meningitidis is the main cause of bacterial meningitis, but genomic studies have suggested that alleles of nitrate reductase, factor-H biding protein and capsule are associated with N. meningitidis isolation in genitourinary infections. Transmission from the oropharynx to the urethra through orogenital contact in unprotected oral sex has been widely proven. N. meningitidis prevalence as the cause of the urethritis is low, and the asymptomatic carriers in the urethra are extremely rare. PCR is a method for the N. gonorrhoeae and C.trachomatis diagnoses, but it does not detect N. meningitidis. The gonorrhoea diagnosis is based on an increased number of polymorphonuclear cells, with intracellular gram-negative diplococci in Gram' stain of urethral discharge. In our case, the gram-negative diplococcus seen in the stain was a meningococcus. Urethritis due to N. meningitidis is indistinguishable from the secondary to N. gonorrhoeae, mimicking it even microscopically, only the epidemiology varies. The conventional bacteriological culture continues to be essential for a correct diagnosis.

How to Identify Invasive Candidemia in ICU-A Narrative Review.

The incidence of invasive fungal infection in ICUs has increased over time, and Candida spp. is the most common cause. Critical care patients are a particular set of patients with a higher risk of invasive fungal infections; this population is characterized by extensive use of medical devices such as central venous lines, arterial lines, bladder catheters, hemodialysis and mechanical intubation. Blood cultures are the gold standard diagnosis; still, they are not an early diagnostic technique. Mannan, anti-mannan antibody, 1,3-ß-D-glucan, Candida albicans germ tube antibody, Vitek 2, PNA-FISH, MALDI-TOF, PCR and T2Candida panel are diagnostic promising microbiological assays. Scoring systems are tools to distinguish patients with low and high risk of infection. They can be combined with diagnostic tests to select patients for pre-emptive treatment or antifungal discontinuation. Candidemia is the focus of this narrative review, an approach to contributing factors and diagnosis, with an emphasis on critical care patients.

Haemophilus urethritis in males: A series of 30 cases.

INTRODUCTION AND OBJECTIVES: Pathogens such as Haemophilus spp. have been associated with non-gonococcal urethritis, but their role is unproven. To describe the clinical characteristics and therapeutic outcomes in male patients diagnosed with Haemophilus spp. urethritis. METHODS: We carried out a retrospective study of all patients who presented to our hospital (in either the emergency department or the outpatient clinic) between July 2016 and April 2018 in whom Haemophilus spp. was isolated in the urethral samples. We enrolled 30 men with Haemophilus spp.-positive urethritis, including coinfections with Neisseria gonorrhoeae and Chlamydia trachomatis. Clinical, laboratory, demographic, and behavioral data were obtained by reviewing medical histories. RESULTS: The mean age of the patients was 36.6 years (range 21-87). Seventeen patients (63%) reported being exclusively heterosexual. Three patients (10%) were HIV infected, all of them with an undetectable viral load. The most common clinical presentation was mucopurulent urethral discharge, in 13 patients (43%). The antibiotic treatment achieved a complete clinical resolution in 73%. CONCLUSIONS: Haemophilus urethritis affected men regardless of their sexual orientation or HIV status. Unprotected oral sex may play a role in its transmission. The limitations of the study preclude verification of the pathogenic role of Haemophilus spp. in acute urethritis, but clinical response after antibiotic treatment suggests that Haemophilus spp. can play such a role.

[Evolution of patient colonization with methicillin-resistant Staphylococcus aureus in a skilled nursing facility]. / Evolución de la colonización por Staphylococcus aureus resistente a meticilina en un hospital de media y larga estancia.

BACKGROUND AND OBJECTIVE: Methicillin-resistant Staphylococcus aureus (MRSA) asymptomatic colonization is common in long-term care facilities, but the burden of symptomatic infection appears to be low. It is not usually known whether a patient is colonized at the time of admission to the geriatric facility. Our purpose was to determine the prevalence, characteristics and factors associated with MRSA colonization on admission, and the cumulative incidence of colonization over the following 6 months. PATIENTS AND METHOD: Longitudinal and prospective study conducted over a 6-month period. All patients were screened at admission using nasal and ulcers swabs within the first 24h. Patients were screened also at the end of the study to assess carrier status. RESULTS: The prevalence of MRSA colonization was 7.6% at the entry (25 patients). In the multivariate analysis, advanced age, recent use of antibiotics, prior colonization by MRSA, and peripheral vascular disease were independent risk factors for colonization at admission. With standard precautions, the 6-month cumulative incidence of MRSA colonization was 4.2%. CONCLUSIONS: In our long-term care facility, MRSA colonization at the time of admission was frequent. Few patients were colonized during the study and no episodes of infection were reported. Probably, standard precautions, including hand washing and appropriate barrier procedures during the care of wounds, are the most useful control measures.

Molecular characterization of OXA-48 carbapenemase-producing Klebsiella pneumoniae strains after a carbapenem resistance increase in Catalonia.

INTRODUCTION: To characterize OXA-48 carbapenemase-producing Klebsiella pneumoniae strains isolated after an increase in carbapenem resistance in Catalonia. METHODOLOGY: K. pneumoniae identification, antimicrobial susceptibility studies, the Modified Hodge Test method, amplification of antimicrobial resistance genes (against ß-lactamases, quinolones and aminoglycosides), molecular typing (by PFGE and MLST), conjugation assays, plasmid characterization (PBRT-PCR and Southern blot), a description of mobile genetic elements and statistical analysis were done. RESULTS: OXA-48 was the only carbapenemase detected, with a prevalence of 1.9%. The blaOXA-48 gene was located in an IncL conjugative plasmid of 62kb and integrated into the transposons Tn1999.2 (91.7%) or Tn1999.1. Five PFGE profiles (A to E) were found, which exactly matched the MLST: ST101, ST17, ST1233, ST14 and ST405, respectively. ST1233 is described here for the first time. K. pneumoniae OXA-48-producing strains were also CTX-M-15 carriers, some producing OXA-1 and TEM-1 penicillinases. The acquired qnrB66 and qnrB1 and aac(3')-IIa, aac(6')-Ib genes were also identified. CONCLUSION: The K. pneumoniae ST405 clone has played an important role in the growing prevalence of OXA-48 in Catalonia. All clones described preserved the blaOXA-48 genetic environment and mobile genetic elements (Tn1999). Notably, the three strains with minor sequence types in this study are not multiresistant strains. These strains are expanding in elderly patients (average age of 76 years) with serious underlying diseases, mainly women (61.2%).

Characterisation of the CTX-M-15-encoding gene in Klebsiella pneumoniae strains from the Barcelona metropolitan area: plasmid diversity and chromosomal integration.

The localisation and genetic organisation of bla(CTX-M-15) were studied in 37 CTX-M-15-producing Klebsiella pneumoniae isolates collected from 2005 to 2008 within the Barcelona metropolitan area. Polymerase chain reaction (PCR)-based replicon typing and Southern hybridisations were used to identify the bla(CTX-M-15) location. The genetic environment was analysed by PCR mapping and sequencing, and transferability of bla(CTX-M-15) was evaluated by conjugation and transformation assays. The majority of the 37 isolates carried bla(CTX-M-15) in a plasmid location, frequently associated with the aac(6')-Ib-cr gene. Plasmids encoding bla(CTX-M-15) carried three distinct replicons, i.e. IncFII, IncR and IncFIIk, the latter two not having been described previously in association with bla(CTX-M-15). Several of these plasmids were not self-transferable. Furthermore, in all isolates belonging to sequence type ST-1, bla(CTX-M-15) was found integrated into the K. pneumoniae chromosome. In all the studied isolates, the mobile element ISEcp1 was found upstream of bla(CTX-M-15), whereas IS26 was found inserted within ISEcp1 in several isolates, in previously unreported positions. In conclusion, these findings indicate that among K. pneumoniae strains isolated in the Barcelona metropolitan area, bla(CTX-M-15) is associated with diverse genetic elements, including the IncR and IncFIIk replicons, as reported for the first time here, and the chromosome.

Prophylactic intracameral cefazolin after cataract surgery: endophthalmitis risk reduction and safety results in a 6-year study.

PURPOSE: To assess the use of intracameral cefazolin in preventing endophthalmitis in cataract surgery. SETTING: Ophthalmology Department, L'Hospitalet de Llobregat, Barcelona, Spain. METHODS: This study was of phacoemulsification procedures performed from January 2002 to December 2007. In January 2004, intracameral cefazolin given at the end of the surgery was added to the prophylaxis protocol of cataract surgery. The cumulative incidence of postoperative endophthalmitis before and after the addition of intracameral cefazolin was compared. RESULTS: During the study period, 18579 phacoemulsification procedures were performed. In the 2-year period before introduction of intracameral cefazolin prophylaxis, 25 cases of endophthalmitis were diagnosed in 5930 surgeries, leading to a cumulative incidence of 0.422% (95% confidence interval [CI], 0.279%-0.613%). After the introduction of cefazolin, 6 cases of endophthalmitis were diagnosed in 12649 surgeries, an incidence of 0.047% (95% CI, 0.019%-0.099%). When only microbiologically proven cases were considered, the cumulative endophthalmitis incidence was 0.388% (95% CI, 0.252%-0.572%) in the first study period and 0.032% (95% CI, 0.010%-0.076%) in the second study period (P<.0000001). The relative risk for presenting with endophthalmitis in the first study period compared with the second period was 8.89 (95% CI, 3.65-21.65). CONCLUSIONS: A 2.5 mg/0.1 mL intracameral bolus of cefazolin provided excellent prophylactic effectiveness, with a reduction in the incidence of endophthalmitis from 0.422% to 0.047%, corresponding to a relative risk reduction of 88.7% (95% CI, 72.6%-95.4%). Cefazolin fulfills international recommendations on antimicrobial prophylaxis for surgical site infections and is easier to obtain in developing countries.

Epidemiology, risk factors, and prognosis of Candida parapsilosis bloodstream infections: case-control population-based surveillance study of patients in Barcelona, Spain, from 2002 to 2003.

Candida parapsilosis has emerged as an important yeast species causing fungemia. We describe the incidence and epidemiology of C. parapsilosis fungemia. Data from active population-based surveillance in Barcelona, Spain, from January 2002 to December 2003 were analyzed. We focused on 78 episodes of C. parapsilosis fungemia, and we compared them with 175 Candida albicans controls. C. parapsilosis accounted for 23% of all fungemias. The annual incidences were 1 episode per 10(5) patients, 1.2 episodes per 10(4) discharges, and 1.7 episodes per 10(5) patient days. All isolates but one (99%) were fluconazole susceptible. Seventy-two isolates (92%) were inpatient candidemias. Forty-two episodes (51%) were considered catheter-related fungemia, 35 (45%) were considered primary fungemia, and 3 (4%) were considered secondary fungemia. Risk factors for candidemia were vascular catheterization (97%), prior antibiotic therapy (91%), parenteral nutrition (54%), prior surgery (46%), prior immunosuppressive therapy (38%), malignancy (27%), prior antifungal infection (26%), transplant recipient (16%), neutropenia (12%), and prior colonization (11%). Multivariate analysis of the differential characteristics showed that the factors that independently predicted the presence of C. parapsilosis fungemia were neonate patients (odds ratio [OR], 7.5; 95% confidence interval [CI], 2.1 to 26.8; P = 0.002), transplant recipients (OR, 9.2; 95% CI, 1.9 to 43.3; P = 0.005), patients with a history of prior antifungal therapy (OR, 5.4; 95% CI, 1.8 to 15.9; P = 0.002), and patients who received parenteral nutrition (OR, 2.2; 95% CI, 1.09 to 4.6; P = 0.028). The overall mortality rate was lower than that associated with C. albicans candidemia (23% versus 43%; P < 0.01). In summary, C. parapsilosis was responsible for 23% of all candidemias and was more frequent in neonates, in transplant recipients, and in patients who received parenteral nutrition or previous antifungal therapy, mainly fluconazole. The mortality rate was lower than that associated with C. albicans fungemia.

Levofloxacin treatment failure in Haemophilus influenzae pneumonia.

We describe the first case of failure of oral levofloxacin treatment of community-acquired pneumonia caused by Haemophilus influenzae. The strain showed cross-resistance to fluoroquinolones and carried four mutations in quinolone resistance-determining regions of DNA gyrase and topoisomerase IV genes.