Emerging non-13-valent pneumococcal conjugate vaccine (PCV13) serotypes causing adult invasive pneumococcal disease in the late-PCV13 period in Spain.

OBJECTIVE: An early reduction of adult invasive pneumococcal disease (IPD) was observed after the 13-valent pneumococcal conjugate vaccine (PCV13) introduction for children in Spain. We analysed the epidemiology of adult IPD in the late-PCV13 period. METHODS: This was a prospective multicentre study of adult IPD involving six hospitals. Strains were serotyped, genotyped and studied for antimicrobial susceptibility. The late-PCV13 period was compared with the pre- and early-PCV13 periods. RESULTS: A total of 2197 episodes were collected-949 in 2008-2009, 609 in 2012-2013 and 639 in 2015-2016. The initial decrease of IPD observed (from 12.3/100 000 to 8.1/100 000; 2008-2009 versus 2012-2013) plateaued in 2015-2016 (8.3/100 000). IPD due to PCV13 serotypes decreased (from 7.7 to 3.5 to 2.3/100 000; p < 0.05), whereas IPD caused by non-PCV13 serotypes increased (from 4.5 to 4.6 to 6.0/100 000; p < 0.05). The most frequent serotypes in the late-PCV13 period were: 8 (15.1%), 3 (10.5%), 12F (7.9%) and 9N (5.4%). These serotypes were related to major genotypes: CC53 (59.8%) and CC404 (30.4%) for serotype 8, CC180 (64.1%) and CC260 (28.1%) for serotype 3, CC989 (91.7%) for serotype 12F and CC67 (84.8%) for serotype 9N. Penicillin-non-susceptibility (21.2%) was associated with serotypes 11A (CC156), 14 (CC156) and 19A (CC320), and macrolide-resistance was related to serotypes 24F and 19A. Rates of pneumococcal meningitis remained stable throughout the periods (ranges 0.9, 0.8 and 1.0/100 000). CONCLUSIONS: The initial decrease of adult IPD observed after PCV13 introduction for children has been balanced by the rise of non-PCV13 serotypes. The spread of antibiotic-resistant lineages related to non-PCV13 serotypes (11A and 24F) could be a threat for the treatment of serious pneumococcal diseases.

Evolution over a 15-year period of the clinical characteristics and outcomes of critically ill patients with severe community-acquired pneumonia / Evolución durante un período de 15 años de las características clínicas y resultados de los pacientes críticamente enfermos con neumonía comunitaria grave

OBJECTIVES: To study the characteristics and outcomes of patients in the ICU with severe community-acquired pneumonia (SCAP) over a 15-year surveillance period. METHODS: We conducted a retrospective cohort study of episodes of SCAP, and assessed the epidemiology, etiology, treatment and outcomes of patients admitted to the ICU, comparing three periods (1999-2003, 2004-2008 and 2009-2013). RESULTS: A total of 458 patients were diagnosed with SCAP. The overall cumulative incidence was 37.4 episodes/1000 admissions, with a progressive increase over the three periods (P < 0.001). Patients fulfilling the two major IDSA/ATS criteria at admission increased from 64.2% in the first period to 82.5% in the last period (P = 0.005). Streptococcus pneumoniae was the prevalent pathogen. The incidence of bacteremia was 23.1%, and a progressive significant reduction in overall incidence was observed over the three periods (P = 0.02). Globally, 91% of the patients received appropriate empiric antibiotic treatment, increasing from 78.3% in the first period to 97.7% in the last period (P < 0.001). Combination antibiotic therapy (betalactam+macrolide or fluoroquinolone) increased significantly from the first period (61%) to the last period (81.3%) (P < 0.001). Global ICU mortality was 25.1%, and decreased over the three periods (P = 0.001). CONCLUSIONS: Despite a progressively higher incidence and severity of SCAP in our ICU, crude ICU mortality decreased by 18%. The increased use of combined antibiotic therapy and the decreasing rates of bacteremia were associated to improved patient prognosis

OBJETIVOS: Estudiar las características y el pronóstico de los pacientes con neumonía grave adquirida en la comunidad ingresados en una unidad de cuidados intensivos (UCI) durante un período de 15 años. MATERIAL Y MÉTODOS: Estudio retrospectivo de una cohorte de pacientes con neumonía grave comunitaria en los que se analizó la evolución en la epidemiología, etiología, tratamiento y pronóstico durante un período de 15 años, comparando 3 períodos (1999-2003, 2004-2008 y 2009-2013). RESULTADOS: Un total de 458 pacientes fueron diagnosticados de neumonía. La incidencia media global durante el período estudiado fue de 37,4 episodios/1.000 ingresos, encontrándose un incremento progresivo durante los 3 períodos estudiados (p < 0,001). Los pacientes que cumplían con los 2 criterios mayores de neumonía grave de la ATS/IDSA aumentaron de un 64,2% en el primer período a un 82,5% en el último período (p = 0,005). Streptococcus pneumoniae fue el microorganismo más frecuente aislado. La incidencia de bacteriemia fue del 23,1%, encontrándose una reducción significativa y progresiva en la incidencia a lo largo de los 3 períodos (p = 0,02). El 91% de los pacientes recibió tratamiento antibiótico empírico apropiado, encontrándose un incremento entre el primer y el último período del 78,3% al 97,7% (p < 0,001). El tratamiento combinado (betalactámico+macrólido o quinolona) aumentó de un 61% en el primer período a un 81,3% en el último (p < 0,001). La mortalidad en la unidad de cuidados intensivos durante todo el período fue del 25,1%, encontrándose una disminución progresiva durante los 3 períodos (p=0,001). CONCLUSIONES: A pesar de un incremento progresivo en la incidencia y gravedad de las neumonías ingresadas en unidad de cuidados intensivos, la mortalidad se redujo en un 18%. El incremento en la utilización de tratamiento combinado y la disminución en la incidencia de bacteriemia se asociaron a una mejoría en el pronóstico

Evolution over a 15-year period of the clinical characteristics and outcomes of critically ill patients with severe community-acquired pneumonia.

OBJECTIVES: To study the characteristics and outcomes of patients in the ICU with severe community-acquired pneumonia (SCAP) over a 15-year surveillance period. METHODS: We conducted a retrospective cohort study of episodes of SCAP, and assessed the epidemiology, etiology, treatment and outcomes of patients admitted to the ICU, comparing three periods (1999-2003, 2004-2008 and 2009-2013). RESULTS: A total of 458 patients were diagnosed with SCAP. The overall cumulative incidence was 37.4 episodes/1000 admissions, with a progressive increase over the three periods (P<0.001). Patients fulfilling the two major IDSA/ATS criteria at admission increased from 64.2% in the first period to 82.5% in the last period (P=0.005). Streptococcus pneumoniae was the prevalent pathogen. The incidence of bacteremia was 23.1%, and a progressive significant reduction in overall incidence was observed over the three periods (P=0.02). Globally, 91% of the patients received appropriate empiric antibiotic treatment, increasing from 78.3% in the first period to 97.7% in the last period (P<0.001). Combination antibiotic therapy (betalactam+macrolide or fluoroquinolone) increased significantly from the first period (61%) to the last period (81.3%) (P<0.001). Global ICU mortality was 25.1%, and decreased over the three periods (P=0.001). CONCLUSIONS: Despite a progressively higher incidence and severity of SCAP in our ICU, crude ICU mortality decreased by 18%. The increased use of combined antibiotic therapy and the decreasing rates of bacteremia were associated to improved patient prognosis.

Differential time to positivity of blood cultures: A valid method for diagnosing catheter-related bloodstream infections in the intensive care unit

Purpose: The validation in critical patients with short-term catheters of a method for diagnosing catheter-related bloodstream infection (CR-BSI), based on the differential time to positivity (DTP) of blood cultures. Methods: Patients suspected of having CR-BSI were included. Two peripheral vein blood cultures and a catheter hub blood culture were simultaneously carried out. The responsible catheter was removed and tip cultured. Times to positivity of all blood cultures were automatically registered. CR-BSI was diagnosed when all the cultures were positive for the same microorganism and DTP>120min. This diagnosis was compared with the one obtained using the standard method. Results: 226 cases suspected of CR-BSI were analyzed during a 20-month period. A total of 19 removed catheters were associated with CR-BSI. Seven cases of polymicrobial cultures (4 with CR-BSI) were discarded from the final analysis due to the impossibility of determining the time to positivity for each individual microorganism. Using the DTP method, 12 out of 15 CR-BSI cases were diagnosed (sensitivity 80%, specificity 99%, PPV 92%, NPV 98%). In a ROC curve, we found a cut-off value of 17.7h in positivity of hub blood cultures that may be useful for diagnosing CR-BSI. Conclusion: DTP can be a valid method for CR-BSI diagnosis in critically ill patients, avoiding unnecessary catheter withdrawal (AU)

Objetivos: La validación, en pacientes críticos con catéteres de corta duración, de un método diagnostico de la bacteriemia asociada a catéter (BAC) basado en la diferencia en el tiempo de positivización (DTP) de hemocultivos. Material y Métodos: Se incluyeron pacientes con sospecha de BAC en los que se realizaron 2 hemocultivos de sangre periférica y un hemocultivo a través de la luz distal del catéter sospechoso, antes de la retirada y cultivo de la punta del mismo. Se registraron automáticamente los tiempos de positivización de todos los hemocultivos. Diagnosticamos BAC cuando todos los hemocultivos fueron positivos para el mismo microorganismo y el DTP >120 minutos. La exactitud de este método diagnóstico fue comparada con la obtenida mediante el método estandar. Resultados: Se analizaron 226 casos de sospecha de BAC durante 20 meses. En 19 de ellos se diagnosticó BAC mediante el método estandar. En 7 casos los hemocultivos fueron polimicrobianos (4 de ellos asociados a BAC) por lo que tuvieron que ser descartados para el analisis final dada la imposibilidad de determiner el tiempo de positivización de cada microorganismo por separado. Siguiendo el método basado en el DTP, 12 de los 15 casos de BAC fueron diagnosticados correctamente (sensibilidad 80%, especificidad 99%, VPP 92%, VPN 98%). En una curva ROC, encontramos un punto de corte de 17.7 horas en el tiempo de positivización del hemocultivo a través de catéter que puede ser útil para el diagnóstico de BAC. Conclusion: La DTP puede ser un método valido para el diagnostico de BAC monobacteriana en pacientes críticos con catéteres de corta duración, evitando la retirada innecesaria de catéteres (AU)

Differential time to positivity of blood cultures: a valid method for diagnosing catheter-related bloodstream infections in the intensive care unit.

PURPOSE: The validation in critical patients with short-term catheters of a method for diagnosing catheter-related bloodstream infection (CR-BSI), based on the differential time to positivity (DTP) of blood cultures. METHODS: Patients suspected of having CR-BSI were included. Two peripheral vein blood cultures and a catheter hub blood culture were simultaneously carried out. The responsible catheter was removed and tip cultured. Times to positivity of all blood cultures were automatically registered. CR-BSI was diagnosed when all the cultures were positive for the same microorganism and DTP≥120 min. This diagnosis was compared with the one obtained using the standard method. RESULTS: 226 cases suspected of CR-BSI were analyzed during a 20-month period. A total of 19 removed catheters were associated with CR-BSI. Seven cases of polymicrobial cultures (4 with CR-BSI) were discarded from the final analysis due to the impossibility of determining the time to positivity for each individual microorganism. Using the DTP method, 12 out of 15 CR-BSI cases were diagnosed (sensitivity 80%, specificity 99%, PPV 92%, NPV 98%). In a ROC curve, we found a cut-off value of 17.7 h in positivity of hub blood cultures that may be useful for diagnosing CR-BSI. CONCLUSION: DTP can be a valid method for CR-BSI diagnosis in critically ill patients, avoiding unnecessary catheter withdrawal.

Evaluation of the VersaTREK system compared to the Bactec MGIT 960 system for first-line drug susceptibility testing of Mycobacterium tuberculosis.

The aim of this study was to evaluate the reliability of the VersaTREK system for Mycobacterium tuberculosis drug susceptibility testing compared with results obtained with the Bactec MGIT 960 system. A total of 67 strains were evaluated. Overall agreement was at 98.5%. Kappa indexes were 1.0 for isoniazid, rifampin, and ethambutol, 0.937 for pyrazinamide, and 0.907 for streptomycin. The VersaTREK system is validated for M. tuberculosis drug susceptibility testing.

Influence of pneumococcal serotype group on outcome in adults with bacteraemic pneumonia.

The influence of infecting serotype group on outcome in bacteraemic pneumococcal pneumonia remains unclear. We performed a prospective, 10-yr observational study in an 800-bed teaching hospital. 299 adults diagnosed with pneumonia whose blood cultures showed growth of Streptococcus pneumoniae were included in the study. High invasive disease potential (H) serotypes included serotypes 1, 5 and 7F, which served as a reference category, were compared with low invasive disease potential (L) serotypes (3, 6A, 6B, 8, 19F, and 23F) and other (O) serotypes (non-H, non-L). The influence on outcome was determined for each group of serotypes after adjusting for underlying conditions and severity of illness at admission. Overall, 30-day mortality was 11%. H serotypes (n = 93) infected primarily younger people and presented a higher risk of complicated parapneumonic effusion or empyema (17.2 versus 5.1%; p = 0.01), with lower mortality (3.2%). The isolation of L serotypes (n = 78) was an independent risk factor for 30-day mortality (OR 7.02, 95% CI 1.72-28.61), as were Charlson score (OR 1.30, 95% CI 1.08-1.58), alcohol abuse (OR 3.99, 95% CI 1.39-11.39) and severity of illness measured by American Thoracic Society (ATS)/Infectious Diseases Society of America (IDSA) criteria (OR 4.80, 95% CI 1.89-12.13). A vaccination strategy including serotypes 3, 6A, 6B, 8, 19F and 23F may improve survival in adults.

Prospective randomized trial of 3 antiseptic solutions for prevention of catheter colonization in an intensive care unit for adult patients.

OBJECTIVE: To compare the effectiveness for prevention of central venous and arterial catheter colonization of 3 skin antisepsis with 1 of 3 antiseptic solutions: 10% aqueous povidone iodine (aqueous PI), 2% aqueous chlorhexidine gluconate (aqueous CG), and 0.5% alcoholic chlorhexidine gluconate (alcoholic CG). DESIGN: Prospective, randomized controlled trial. SETTING: Intensive care unit in a teaching hospital. METHODS: Patients were randomly assigned to 1 of the 3 skin antisepsis groups. The distal tips of catheters were semiquantitatively cultured when the catheters were no longer necessary or if there was a suspicion of catheter-related infection. Rates of catheter colonization, catheter-related sepsis, and catheter-related bacteremia were compared among the 3 groups. RESULTS: A total of 631 catheters were included in the study (194 from the aqueous PI group, 211 from the aqueous CG group, and 226 from the alcoholic CG group). The incidence of catheter colonization was significantly lower in the alcoholic CG than in the aqueous PI group (14.2% vs 24.7%; relative risk, 0.5 [95% confidence interval, 0.3-0.8; P < .01]); it was also significantly lower in the aqueous CG group than in the aqueous PI group (16.1% vs 24.7%; relative risk, 0.6 [95% confidence interval, 0.4-0.9; P = .03]). There were no significant differences between the aqueous CG and the alcoholic CG groups. Incidences of catheter-related bacteremia were similar for all 3 groups. The aqueous and alcoholic CG solutions were superior to the aqueous PI solution in preventing catheter colonization due to gram-positive bacteria. CONCLUSIONS: The aqueous and alcoholic CG solutions for cutaneous antisepsis were similarly effective in preventing colonization of central venous catheters and arterial catheters. Both had significantly lower incidences of colonization than did the aqueous PI solution; this effect seems to be related to the CG solutions' more efficacious prevention of colonization with gram-positive bacteria.

Bloodstream infections in adults: importance of healthcare-associated infections.

OBJECTIVE: The objective of this study was to evaluate the characteristics of bloodstream infections occurring among outpatients having recent contact with the health care system compared to hospital and community-acquired infections. METHODS: Prospective observational cohort study of adult patients with bloodstream infections at three teaching hospitals. Bloodstream infection was defined as hospital-acquired if the first positive blood culture was performed more than 48 h after admission. Other bloodstream infections were classified as healthcare-associated or community-acquired. RESULTS: A total of 1157 episodes of bloodstream infections were studied; 581 (50.2%) were community-acquired, 295 (25.5%) were hospital-acquired, and 281 (24.3%) were health care-associated. Of the 281 health care-associated bloodstream infections, 68 (24%) occurred in patients residing in a nursing home, 104 (37%) in patients receiving intravenous therapy, health care at home, chemotherapy or attending dialysis, and 169 (60%) in patients hospitalized during the 90 days before their bloodstream infection (some patients belonged to more than one risk category). The highest prevalence rate of MRSA infections occurred in healthcare-associated infections (5%) (p<0.001). A significantly higher mortality rate was seen in the group with healthcare-associated infections (27.5%) than in community-acquired infections (10.4%) (p<0.001). CONCLUSIONS: Our results confirm that healthcare-associated bloodstream infections show important differences from community-acquired bloodstream infections and suggest that empirical antibiotic therapy should be similar to hospital-acquired bloodstream infections, taking into account the epidemiologic characteristics of each region.

[An outbreak of gram-negative bacteremia (GNB), especially enterobacter cloacae, in patients with long-term tunnelled haemodialysis catheters]. / Brote epidémico de bacteriemias por bacilos gram-negativos (BGN), especialmente enterobacter cloacae, en una unidad de hemodiálisis, en pacientes portadores de catéteres permanentes tunelizados.

Vascular access through a venous catheter for haemodialysis is associated with increased risk of thrombosis, central venous stenosis, short access survival and inadequate dialysis. The most important catheter-related complications, which determine method survival, are infection and dysfunction. In particular, infectious episodes are in some studies the leading cause for untimely catheter removal and for catheter-related morbidity but also for morbidity in dialysis patients. Double-lumen central venous catheters used for haemodialysis, are common causes of septicaemia. Most cases are caused by staphylococci. Episodes of gram-negative bacteriemia have been traced to bacterial contamination of water and/or dialysate, errors in dialyzer reprocessing, and improper setup procedures. In this paper, we describe and outbreak of gram-negative bacteremia, firstly E. cloacae, in an outpatients haemodialysis unit, in the patients with long-term tunnelled haemodialysis catheters. We describe the epidemic investigation that we achieved to identify the source of contaminating bacteria and the route by which bacteria gained access to the bloodstream. We prove the contamination by gram-negative bacterium of the water-distribution lines and haemodialysis machines. Moreover, E. cloacae strains isolated from the lines and machines are genotypically identical to the isolated from the patients. Also, we prove that the hands of health care personnel are unintentional carriers. The outbreak was finished when decontamination of dialysis machines was enhanced and dialyzer-priming fluid was modified.

Brote epidémico de bacteriemias por bacilos gram-negativos (BGN), especialmente enterobacter cloacae, en una unidad de hemodiálisis, en pacientes portadores de catéteres permanentes tunelizados / An outbreak of Gram-Negative Bacteremia (GNB), especially Enterobacter Cloacae, in patients with long-term tunnelled haemodialysis catheters

Se considera a los catéteres permanentes tunelizados un acceso vascular para hemodiálisis de segundo orden por la morbilidad más elevada que comportan respecto a otros accesos. Las complicaciones son más numerosas, y se relacionan especialmente con la trombosis parcial que origina disfunción en la diálisis, y con las infecciones. Estas últimas pueden originar bacteriemias, mayoritariamente debidas a cocos gram-positivos, estafilococos sobre todo. Las infecciones por bacilos gram-negativos (BGN) son más excepcionales y su origen, aún hoy en día, no está explicado. En las Unidades de Hemodiálisis, se han descrito brotes epidémicos relacionados con la contaminación accidental del agua tratada, del líquido de diálisis o del material accesorio de las máquinas. Describimos un brote epidémico de bacteriemias por BGN, especialmente Enterobacter cloacae, en la Unidad de Hemodiálisis de pacientes crónicos, en enfermos portadores de catéteres permanentes tunelizados. Detallamos minuciosamente los pasos seguidos para intentar descubrir el origen de las bacteriemias, consiguiendo finalmente demostrar la contaminación por BGN de los accesorios de los monitores de hemodiálisis, y el papel humano como vector transmisor involuntario. Establecemos la relación genética entre las cepas de E. cloacae aisladas en los hemocultivos de los pacientes y en los accesorios contaminados de los monitores de hemodiálisis. Tras el aumento de la desinfección de los accesorios de los monitores de diálisis y el cambio en el modo de cebado de los dializadores, se ha conseguido erradicar las bacteriemias por BGN en nuestra Unidad

Vascular access through a venous catheter for haemodialysis is associated with increased rik of thrombosis, central venous stenosis, short access survival and inadequate dialysis. The most important catheter-related complications, which determine method survival, are infection and dysfunction. In particular, infectious episodes are in some studies the leading cause for untimely catheter removal and for catheter-related morbidity but also for morbidity in dialysis patients. Double-lumen central venous catheters used for haemodialysis, are common causes of septicaemia. Most cases are caused by staphylococci. Episodes of gram-negative bacteriemia have been traced to bacterial contamination of water and/or dialysate, errors in dialyzer reprocessing, and improper setup procedures. In this paper, we describe and outbreak of gram-negative bacteremia, firstly E. cloacae, in an outpatients haemodialysis unit, in the patients with long-term tunnelled haemodialysis catheters. We describe the epidemic investigation that we achieved to identify the source of cantaminating bacteria and the route by which bacteria gained access to the bloodstream. We prove the contamination by gramnegative bacterium of the water-distribution lines and haemodialysis machines. Moreover, E. cloacae strains isolated from the lines and machines are genotypically identical to the isolated from the patients. Also, we prove that the hands of health care personnel are unintentional carriers. The outbreak was finished when decontamination of dialysis machines was enhanced and dialyser-priming fluid was modified

Epidemiology of invasive Streptococcus pneumoniae infections in children in an area of Barcelona, Spain.

AIM: Streptococcus pneumoniae is the most common cause of bacteraemia, pneumonia, sinusitis and acute otitis media. With the advent of conjugate vaccines, there is now the possibility of preventing disease caused by this organism. However, little is known about the epidemiology of invasive pneumococcal disease in children in Spain. The aim of this study was to determine the incidence and the clinical and microbiologic characteristics of invasive pneumococcal disease in Sabadell, an industrial area in the province of Barcelona, Spain. METHODS: From January 1990 to December 2000, the case records of children with pneumococcal invasive disease at Sabadell Hospital were retrospectively (1990-1996) reviewed and prospectively (1997-2000) collected. The hospital serves a population of 61,143 children under 15 y of age, 18,073 children under 4 y of age and 7300 children under 2 y of age. RESULTS: A total of 112 children (54% under 24 mo of age and 93% under 6 y of age) with invasive pneumococcal disease were diagnosed during a period of 11 y. The incidence of invasive pneumococcal disease was 76 per 100,000 for children aged 0 to 24 mo, 45 for children aged 0-48 mo and 16.6 for children aged 0-14 y. Occult bacteraemia was the most common manifestation of invasive pneumococcal disease (66 cases), pneumonia was the second form (34 cases) and meningitis (10 cases) and arthritis (2 cases) were the other clinical manifestations. Of the 105 strains tested, 8.6% were highly penicillin resistant, 37.1% were intermediately penicillin resistant. 16.2% were intermediately cefotaxime resistant and 32.4% were erythromycin resistant. Pneumococci of serogroups 6, 14, 18, 19, 1, 5, 4, 9, 23 and 33 were the most frequently isolated groups (92%) but only 6, 9, 14, 19 and 23 were resistant to penicillin, cefotaxime, or erythromycin. CONCLUSIONS: In this study the incidence of invasive pneumococcal disease was found to be greater than that reported elsewhere in Spain and Europe. Penicillin resistance levels are high but the trend towards increasing penicillin resistance may have ended over the past few years. The currently licensed seven-valent (7-V) pneumococcal conjugate vaccine would cover 78% of cases of invasive pneumococcal in children aged 0-14 y, 80% in children aged 0-24 mo and 100% of cases of penicillin- or cefotaxime-resistant invasive pneumococcal disease.

Association between asymptomatic carriage and sporadic (endemic) meningococcal disease in an open community.

We analysed a strain collection representative of the overall Neisseria meningitidis population circulating in an open community (46,000 inhabitants, Spain) during an endemic period (30 isolates from patients and 191 from throat cultures of healthy individuals) by both phenotypic and molecular techniques. Almost all patient isolates were assigned to three hyper-virulent lineages (ET-5 complex, ET-37 complex and cluster A4) by both multilocus enzyme electrophoresis (MEE) and pulsed-field gel electrophoresis (PFGE). In contrast, MEE and PFGE assigned 20% and 15% respectively of carrier isolates to the hyper-virulent clones (4% for both methods together). There was also a higher correlation between PFGE and phenotypes associated with virulent clones. These notable differences between the two molecular methods were further observed in more than half the carrier isolates, suggesting that the associations between these strains were distorted by recombination events. However, almost one-third of total endemic strains from symptom-free carriers and almost all patient strains belonged to clones defined by MEE and PFGE, with no known epidemiological connection. These data indicate low transmission and a weak clonal structure for N. meningitidis.

Eleven-year study of recurrent pneumococcal bacteremia.

In order to describe the epidemiology and the clinical and microbiological manifestations of recurrent pneumococcal bacteremia, a long-term study was conducted. Between January 1988 and December 1998, a total of 344 episodes of bacteremia caused by pneumococci was detected in 331 patients. Thirteen (3.9%) of these patients experienced recurrent episodes of pneumococcal bacteremia, and all of them had underlying diseases. In 12 of these patients the recurrence was considered to be a reinfection, and in one patient it was considered to be a relapse. Three patients were found to harbor identical strains in both bacteremic episodes, as determined by pulsed-field techniques. Only hematological neoplasia appeared to be a predisposing factor for recurrent pneumococcal bacteremia.

Prevalence of Haemophilus influenzae pharyngeal carriers in the school population of Catalonia. Working Group on invasive disease caused by Haemophilus influenzae.

The objective of this study was to determine the prevalence of healthy Haemophilus influenzae (Hi) pharyngeal carriers in a representative sample of the Catalonian school population, as well as the factors associated. A two-stage cluster sampling was carried out. Parents were given a questionnaire to collect information on sociodemographic and epidemiological variables. A pharyngeal swab was performed on children when informed consent was given by parents, and was cultured on chocolate agar with 260 microg/ml bacitracin. Of the 1212 children studied, 316 (26%) H. influenzae carriers were detected: 5 (0.4%) serotype b, 1 (0.08%) serotype c, 6 (0.5%) serotype e, 5 (0.4%) serotype f, and 299 (24.7%) non-typable. Age, gender and geographical location were the only variables associated with H. influenzae carrier status. The prevalence of non-typable H. influenzae carriers was similar to that of studies carried out in other countries, while that of serotype b carriers was similar to the remainder of H. influenzae capsulates, and lower than that described in previous studies. These data are in accordance with the low incidence of the disease observed in our context, although the possibility that the vaccine coverage may have affected the results of this study cannot be dismissed.

Prevalence of Haemophilus influenzae carriers in the Catalan preschool population. Working Group on Invasive Disease Caused by Haemophilus influenzae.

This study was designed to determine the prevalence of healthy Haemophilus influenzae carriers in a random sample of the preschool population in Catalonia. Oropharyngeal swabs were collected and cultured on chocolate agar supplemented with 260 microg/ml of bacitracin. Four hundred two of the 734 (54.8%) children studied were detected as Haemophilus influenzae carriers: 7 (0.9%) carried serotype b, 14 (1.9%) serotype e, 6 (0.8%) serotype f, and 375 (51%) carried nontypable strains. The results show that, although the prevalence of Haemophilus influenzae carriers is similar to figures reported from other countries, the prevalence of Haemophillus influenzae serotype b carriers is lower and corresponds with the low incidence of invasive disease observed in the Catalan community.

Comparison of conventional ribotyping and PCR-RFLP ribotyping for the analysis of endemic strains of Neisseria meningitidis isolated in a local community over 7 years.

Conventional ribotyping was compared with the PCR amplification of the intergenic spacer region between 16S and 23S rRNA genes (PCR-RFLP ribotyping) when applied to the subtyping of sporadic Neisseria meningitidis strains. Thirty isolates out of a total of 36 meningococcal disease cases, reported as having occurred in a particular community over a 7-year endemic period, were analyzed by each of the methods. Only ribotyping with three restriction enzymes (EcoRI, ClaI and XhoI) gave acceptable discriminatory power for short-term epidemiological purposes. We conclude that conventional ribotyping is a suitable method for typing sporadic meningococcal strains and that it cannot be replaced by the more straightforward PCR-RFLP ribotyping method.

[Evaluation of the polymerase chain reaction technique for the diagnosis of meningitis caused by Neisseria meningitidis and Haemophilus influenzae]. / Valoración de la técnica de reacción en cadena de la polimerasa para el diagnóstico de las meningitis causadas por N. meningitidis y H. influenzae. Grupo de Trabajo de la Enfermedad Invasiva por H. influenzae.

BACKGROUND: Bacterial meningitis is a severe infection of the central nervous system (CNS), most frequently caused by Neisseria meningitidis in our setting. Microbiologic diagnosis of bacterial meningitis is not enough sensitive because its efficiency can be affected by the therapeutic regimen given to the patient. Polymerase chain reaction (PCR) can provide a more sensitive diagnosis and allow us to get an earlier result. OBJECTIVES: To assess the sensitivity and specificity of a PCR technique for the diagnosis of meningitis caused by N. meningitidis and Haemophilus influenzae. MATERIAL AND METHODS: Ninety-six patients who were attended because of suspected bacterial meningitis on the Hospital de Sant Joan de Déu, Corporació Sanitària Parc Taulí and Hospital de la Santa Creu i Sant Pau, and had negative results by conventional laboratory methods, were selected for the study. A total of 99 cerebrospinal fluid samples (CSF) were obtained and evaluated for PCR. DNA extracts of the CSF samples were amplified by universal primers. Amplification products were hybridized with specific probes for Haemophilus genus and N. meningitidis. Positive and negative controls were included to asses the reliability of PCR. RESULTS: Eight of the 99 CSF samples (8%) were positive by PCR and subsequent hybridization with the specific probe of N. meningitidis. None of the amplicons hybridized with the probe of Haemophilus genus. Thirteen percent of the patients (8/59) with clinical suspicious of non-neonatal sepsis or meningitis were diagnosed by PCR, amongst them, 36% of the cases (4/11) with initial diagnosis of meningococcal sepsis or meningitis. CONCLUSIONS: The sensitivity and the specificity of the PCR technique afford a complementary method to conventional ones, in special for the diagnosis of meningococcal meningitis in the group of pediatric patients.