Comparison of PCR-based methods for the simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in clinical samples

Abstract Background Several in-house PCR-based assays have been described for the detection of bacterial meningitis caused by Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae from clinical samples. PCR-based methods targeting different bacterial genes are frequently used by different laboratories worldwide, but no standard method has ever been established. The aim of our study was to compare different in-house and a commercial PCR-based tests for the detection of bacterial pathogens causing meningitis and invasive disease in humans. Methods A total of 110 isolates and 134 clinical samples (99 cerebrospinal fluid and 35 blood samples) collected from suspected cases of invasive disease were analyzed. Specific sets of primers frequently used for PCR-diagnosis of the three pathogens were used and compared with the results achieved using the multiplex approach described here. Several different gene targets were used for each microorganism, namely ctrA, crgA and nspA for N. meningitidis, ply for S. pneumoniae, P6 and bexA for H. influenzae. Results All used methods were fast, specific and sensitive, while some of the targets used for the in-house PCR assay detected lower concentrations of genomic DNA than the commercial method. An additional PCR reaction is described for the differentiation of capsulated and non-capsulated H. influenzae strains, the while commercial method only detects capsulated strains. Conclusions The in-house PCR methods here compared showed to be rapid, sensitive, highly specific, and cheaper than commercial methods. The in-house PCR methods could be easily adopted by public laboratories of developing countries for diagnostic purposes. The best results were achieved using primers targeting the genes nspA, ply, and P6 which were able to detect the lowest DNA concentrations for each specific target.

Emergencia de la cepa W135 causante de enfermedad meningocócica invasora en Chile 2012 / Emergence of W135 meningococcal serogroup in Chile during 2012

The epidemiologic behavior of the Invasive Meningococcal Disease (IMD) in Chile has changed. At the end of 2011, the W135 serogroup belonging to the hypervirulent clone ST-11 emerged. It affected diverse countries of the world, after the Mecca pilgrimage in 2000. In Chile, there have been 133 IMD cases during 2012. These figures represent an incidence of 0.7 per 100,000 inhabitants, which is 30% higher than expected. Eighty eight percent of cases were confirmed by the National Reference Laboratory at the Chilean Public Health Institute. The serogroup was determined in 103 strains and 58% belonged to the W135 serogroup, surpassing for the first time the B serogroup (37%). The Metropolitan Region concentrated 80% of these cases, and the remaining 20% affected other seven regions of the country. Forty seven percent of cases corresponded to children less than 5 years of age. The predominant clinical presentation of the W135 serogroup was a sepsis in 67% of cases. The fatality ratio of IDM during 2012 was 27%, the highest in the past 20 years. With this information, the Chilean Ministry of Health decreed a sanitary alert and implemented an integrated approach to control and prevent W-135 IDM, denominated "W-135 Action Plan".

Fatal meningococcal meningitis in a HIV-infected patient caused by serogroup C Neisseria meningitidis belonging to the non-hypervirulent clonal complex ST-60 (cc60)

Meningococcal strains belonging to clonal complex cc60 are not associated with hypervirulent lineages and were never reported as causing disease in Latin American countries. This is the first report of a fatal meningitis case caused by a cc60 clonal complex meningococcus in Brazil. Despite the immune-compromised state of the patient, the fatal outcome here described shows the potential pathogenic behavior of strains belonging to this clonal complex and how compromised hosts can be susceptible to meningococcal infections even if the strain is not particularly invasive.

Microbiological epidemiological history of meningococcal disease in Rio de Janeiro, Brazil

The main objectives of the present study were to investigate the clinical and laboratory features of meningococcal disease in the city of Rio de Janeiro, Brazil, during the overlap of 2 epidemics in the 1990s. We conducted a study of a series of cases of meningococcal disease admitted in a Meningitis Reference Hospital. All clinical isolates available were analyzed by means of microbiological epidemiological markers. In 1990, Neisseria meningitidis serogroup B:4,7:P1.19,15, 1.7,1 sulfadiazine-resistant of the ET-5 complex emerged causing epidemic disease. Despite mass vaccination campaign (VaMengoc B+C®), the ET-5 clone remained hyperendemic after the epidemic peaked. In 1993 to 1995, an epidemic of serogroup C belonged to the cluster A4 overlapped, with a significant shift in the age distribution toward older age groups and an increase of sepsis. Serogroup C epidemics are a recurrent problem in Rio de Janeiro, which can be hindered with the introduction of a conjugate vaccine. We hope the data presented here brings useful information to discuss vaccines strategies and early management of suspected cases.

Comparison of dot-ELISA and standard ELISA for detection of Neisseria meningitidis outer membrane complex-specific antibodies

Dot-ELISA using the outer membrane complex antigens of Neisseria meningitidis as a target was standardized for rapid detection of meningococcal-specific antibodies in human serum. We investigated the level of meningococcal-specific IgG, IgA, and IgM in serum using dot-ELISA with outer membrane antigens prepared from Neisseria meningitidis serotype B:4.19:P1.15,3,7,9 (a strain isolated from a Brazilian epidemic). The dot-ELISA is based on the same principles as the standard ELISA and is useful for detection of anti-N. meningitidis B antibodies in serum of patients with meningococcal infections. For the assay, outer membrane complexes (OMCs) were absorbed by nitrocellulose membrane and blocked with a 5 percent skim milk solution. Serum samples were drawn upon hospital admission and during convalescence from patients with meningococcal septicemia, and single samples were drawn from uninfected controls. We retrospectively examined a total of 57 serum samples: 35 from patients infected with N. meningitidis B, 12 from patients infected with Haemophilus influenzae b, and 10 from health individuals. When performed at room temperature, dot-ELISA took approximately four hours to perform, and the optimum antigen concentration was 0.42 µg per dot. The specificity of IgG, IgM, and IgA demonstrates that dot-ELISA using OMCs from N. meningitidis B as a target is suitable for serologic verification of clinically suspected meningococcal disease in patients and for titer determination of antibodies produced during different phases of natural infection. Furthermore, the sensitivity of dot-ELISA was comparable to that of standard ELISA. Overall, dot-ELISA is simple to perform, rapid, and low cost. Further validation of the test as a screening tool is required.

Emergence of neisseria meningitis W135 in Sudan

The Federal Ministry of Health, Sudan has reported 55 suspected cases of meningococcal meningitis including 5 related deaths [CFR: 9%] from South Kordofan State between 31 January to 3 April 2010. The predominant causative strain was identified to be Neisseria meningitides W 135. So far, 5 laboratory samples [CSF] have been tested positive for NMW135 at the National Public Health Laboratory

Prevalence of Neisseria meningitidis carriers in children under five years of age and teenagers in certain populations of Mexico City / Prevalencia de portadores de Neisseria meningitidis en niños menores de cinco años de edad y adolescentes en determinadas poblaciones de la Ciudad de México

OBJECTIVE: To carry out a pilot study to discover the frequency of colonization in healthy children under five years old and teenagers, as well as the distribution of the different N. meningitidis serogroups isolated from nasopharyngeal samples collected from the population under study. MATERIAL AND METHODS: The population included youth between 15 and 19 years old living in social rehabilitation centers (SRC) and university teenagers (UT) as well as children under the age of five attending day care centers (DCC) in Mexico City. Nasopharyngeal exudates were processed using standard microbiological techniques in order to identify Nm, the serogroup, type and subtype of these isolates. RESULTS: A total of 2 310 samples (774 SRC, 800 UT and 736 DCC) were taken. Total prevalence of Nm was 1.6 per cent. In teenagers living in SRC the prevalence was 2.9 percent. The most frequent serogroups were Y (29.7 percent), C (24.3 percent) and B (10.8 percent). CONCLUSIONS: The prevalence of carriers in the study environment was significant (1.6 percent) and similar to countries in which the invasive illness is much more frequent.

OBJETIVO: Realizar un estudio piloto que permita conocer la frecuencia de colonización en niños sanos menores de cinco años y adolescentes, así como la distribución de serogrupos de los aislamientos de Neisseria meningitidis (Nm) obtenidos de la nasofaringe de la población estudiada. MATERIAL Y MÉTODOS: Se incluyeron, jóvenes entre los 15 y 19 años de edad, de centros de readaptación social (CRS) y adolescentes universitarios (AU), así como niños menores de cinco años que asisten a estancias infantiles (EI) en la Ciudad de México. Se tomaron exudados nasofaríngeos y fueron procesados, usando las técnicas microbiológicas clásicas, para identificar Nm, serogrupo, tipo y subtipo de estos aislamientos. RESULTADOS: Se incluyeron un total de 2310 muestras (774 CRS, 800 AU y 736 EI). La prevalencia total de Nm fue 1.6 por ciento y en los adolescentes de CRS fue 2.9 por ciento. Los serogrupos más frecuentes son; Y (29.7 por ciento), C (24.3 por ciento) y B (10.8 por ciento). CONCLUSIÓN: La prevalencia de portadores en nuestro medio fue significativa (1.6 por ciento) y similar a los países en los que la enfermedad invasiva es mucho más frecuente.

Alerts of meningococcal meningitis from countries in EMR

Since the beginning of 2009, WHO received alerts of meningococcal meningitis from two countries in the Region. By end of international epidemiological week 10, Sudan reported 138 cases of meningococcal meningitis including nine related deaths [CFR 6.5%] from Kapoeta and Budi counties, Eastern Equatoria State of southern Sudan. Nisseria Meningitidis, Serogroup A was identified in 8 out of 11 samples tested by Pasteurex, and were isolated in 4 out of 6 samples. In northern Sudan a total of 248 cases including 10 deaths [CFR=4%] from 10 states. Response efforts included risk assessment, case management and targeted immunizations of people aged 2-30 years. Qatar reported 8 isolated cases of meningococcal meningitis between January and 12th February. Three of them were fatal. N. meningitidis sero-group B and W135 were isolated from 6 samples

Meningococcal disease situation in Darfur, Sudan

A total of 180 suspected cases of meningococcal disease including 10 deaths [CFR: 5.5%] were reported from three states of Darfur in Sudan from 01 January to 13 April 2009. Although the laboratory tests has confirmed large predominance of Neisseria meningitides serogroup A as the causative strain of this disease in most parts of Darfur, two suspected cases, reported from West Darfur, have been laboratory confirmed as Neisseria meningitides seerogroup W135. Mass vaccination campaign has been implemented by the Federal Ministry of Health in both south and west Darfur states with support from WHO and other agencies

Diseño y evaluación de una reacción de polimerasa en cadena (RPC) múltiple, para la identificación de bacterias causantes de meningitis aguda en líquido cefalorraquídeo de niños chilenos / Multiplex PCR assay in spinal fluid to identify simultaneously bacterial pathogens associated to acute bacterial meningitis in Chilean children

Background: Acute bacterial meningitis (ABM) is a serious disease that needs rapid diagnosis for an accurate treatment. The most important etiological agents are: Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae type b. Overall pathogen detection rate in patients with ABM in Chile is 83 percent. Aim: To evaluate a Polymerase Chain Reaction (PCR) protocol for simultaneous detection of several pathogens in patients with ABM. Material and methods: We designed and evaluated a multiplex PCR protocol for simultaneous specific genes identifications of S pneumoniae (¡ytA and ply genes), N meningitidis (ctrA, crgA) and H influenzae (bexA) in cerebrospinal fluid (CSF) samples from pediatric patients with suspected diagnosis of ABM. Sensitivity, specificity and minimum detection levels of DNA were determined. Amplifications ofrDNA 16S gene was done to confirm extraction of bacterial DNA. Results: Ninety nine CSF samples were studied, 90 from children with fever and negative CSF culture, and 9 from ABM and positive culture patients. The PCR protocol had a sensitivity of 89 percent, specificity of 100 percent, positive predictive value 100 percent and negative predictive value 99 percent. Conclusions: We observed a high concordance (89 percent) between bacteriological cultures and the PCR protocol results. This diagnostic tool could increase identification of agents in specific settings such as patients previously treated with antibiotics.

Vigilancia de Neisseria meningitidis en Argentina, 1993-2005: distribución de serogrupos, serotipos y serosubtipos causantes de enfermedad invasiva / Surveillance of Neisseria meningitidis in Argentina, 1993-2005: Distribution of serogroups, serotypes and serosubtypes isolated from invasive disease

Neisseria meningitidis es agente causal de enfermedades severas como meningitis, bacteriemia y síndrome de shock séptico. Se presenta la distribución en serogrupos, serotipos y serosubtipos de 2244 aislamientos de N. meningitidis obtenidos de cuadros de meningitis y/o meningococcemia durante el período 1993-2005 y analizados en el Laboratorio Nacional de Referencia del INEI-ANLIS "Dr. Carlos G. Malbrán". Estos aislamientos eran provenientes de 33 hospitales de todo el país, conformados en una red nacional de laboratorios para el estudio de meningitis bacteriana. Durante el período 1993-1995 prevaleció el serogrupo B (66%), mientras que entre los años 1995 y 2001 prevaleció el serogrupo C (65%); a partir de esta fecha se restableció la prevalencia de B. En los últimos 5 años los serogrupos Y y W135 representaron en su conjunto el 15,6%, mientras que hasta el año 2000 no superaron el 4,7%. Se registró mayor diversidad en la distribución de serotipos y serosubtipos dentro del serogrupo B que dentro del serogrupo C. Los aislamientos no subtipables durante todo el período de estudio representaron el 52,8%; este elevado porcentaje evidencia la limitada capacidad de la serología para la determinación de subtipos de meningococo.

Neisseria meningitidis is an important cause of meningitis, bacteremia and septic shock syndrome. We herein present the distribution of serogroups, serotypes and serosubtypes of 2244 isolates of N. meningitidis from patients with meningitis or meningococcemia, received within the period 1993-2005, in the National Reference Laboratory, INEI-ANLIS "Dr. Carlos G. Malbrán", from 33 Argentine hospitals that are included in a National Network devoted to for the study of bacterial meningitis. Between 1993-1995, serogroup B was prevalent (66%) whereas in the period from 1995-2001, serogroup C prevailed (65%). However, following but after that period, the prevalence of serogroup B was recovered. In the last 5 years of the studied period, the serogroups Y and W135 represented as a whole a 15.6% as a whole whereas up to the year 2000 during the first 6 years they accounted for it was of 4.7%. Higher diversity in the distribution of serotypes and serosubtypes was observed within serogroup B. The nonsubtypable isolates throughout the period of study represented the 52.8%, this high percentage demonstrates the limited capacity of the serotyping for the determination of meningococcal/meningococcus subtypes. of meningococco.

Assessment of a two-step nucleic acid amplification assay for detection of Neisseria meningitidis followed by capsular genogrouping

Immediate prevention of meningococcal disease relies in part on the prompt treatment with antibiotics of household and other close contacts of cases; however intervention with effective vaccination relies on identification of serogroup-causing strains. Parenteral antibiotic for patient with suspected meningococcal disease before hospital admission is currently recommended. Laboratory standard methods are hindered by failure to detect bacteria by this medical approach to improve patient prognosis. We assessed two polymerase chain reaction (PCR) assays to detect (crgA) and define the serogroups (siaD, orf-2, and ctrA) of Neisseria meningitidis in 120 cerebrospinal fluid (CSF) samples from positive cases (culture or antigen detection or direct smear). The PCR sensitivity for the identification of N. meningitidis was 100 percent (95 percent confidence interval, CI, 96-100 percent) compared to a sensitivity of 46 percent for culture (95 percent CI 37-55 percent), 61 percent for latex agglutination test (95 percent CI 52-70 percent), and 68 percent for Gram stain (95 percent CI 59-76 percent); PCR specificity was 97 percent (95 percent CI 82-100 percent). PCR correctly identified the serogroups A, B, C, W135, Y, and X in CSF samples with a sensitivity of 88 percent (95 percent CI 80-93 percent); the primer sets were 100 percent specific. The introduction of PCR-based assays shall increase laboratory confirmed cases, consequently enhancing surveillance of meningococcal disease.

Serotyping of Neisseria meningitides in conscripts with meningitis admitted to five military hospital in Tehran between September 2004 and September 2006

Meningococcal meningitis is one of the most feared disorders with high rate morbidity and mortality. There is no exact information of meningococcal meningitis incidence in military personnel. Obligate vaccination of the military with meningococcal vaccine is a strategy with the potency to provide active protection of the conscripts before being dispatched to training centers. Although sporadic cases of meningitis in conscripts occurred The aim of this study was serotyping of the bacterium; Neisseria meningitides isolated from patients with meningitis which admited to five military Hospitals. In this cross- sectional study for determination of meningococcal meningitis serotypes in conscripts submit to five military hospitals the investigation was designed and carried out from September 2003 to September 2006. In this period, 12 cerebrospinal fluids of conscripts with clinical signs and symptoms of meningitis were collected, and the laboratory tests were done. In this study enriched Thayer Martin medium and standard bacteriological methods were used. The sample without bacterial cells or PMNs were centrifuged [10000 xg for 5 min], The precipitate was cultured and direct smear was performed.Isolated bacterial strains was recognized by biochemical tests, and Neisseria meningitides strains were serotyped by specific antiserum. During the study, 12 cases of meningitis in conscripts were seen. Neisseria meningitides was isolated only from 6 patients. Serotyping analysis revealed that 5 strains belong to Serotype C and 1 strain was to serotype B. Neisseria sica was isolated from one patient. In three patients there was no bacteriological evidence. In two patients, consumption of antibiotic before lumbar puncture results to no bacterial growth, but direct smear showed gram negative diplococci. In one patient recurrent meningococcal infection has occurred. Complement components analysis revealed deficiency in C3, C4 and CH50 [the rate was below 80 mg per deciliter]. The results of this study showed that, Neisseria meningitides was isolated from six patients which are all conscripts. It is necessary to find the reasons that why vaccinated conscripts infected by Neisseria meningitides serotype C. Further analysis showed that, five patients had complement deficiency and further research is necessary. However, 6 Neisseria meningitides strains were isolated from meningococcal vaccinated conscripts around the country [Tehran, Shiraz, Yazd and Oslavieh] in the period of study. Based on the findings of this study, effectiveness of the vaccine was desirable and the disease had been controlled in this population group. Because of complements deficiency in patients with meningococcal meningitis, other preventing methods for eradication of disease are necessary

Some epidemiological features of meningococcal meningitis in Saudi Arabia [1994-2004]

Meningococcal disease caused by Nisseria Meningitides repeatedly caused outbreaks worldwide although the majority of cases are sporadic. The aim of this study is to get information about the incidence and pattern of meningococcal meningitis and its serogroups in the period from 1994-2004. The present study is descriptive study conducted during the year 2006 at Saudi Arabia. The total number of cases included in the study were 1038, which represented all confirmed cases of meningococcal meningitis in the period from 1994-2004. Cases were obtained from Ministry of Health Infectious Diseases Department surveillance data base of reportable diseases. Data included: age, sex, nationality, year of the disease occurrence, Nisseria meningitides serogroups, residence of the patient at time of the disease, vaccination status against the disease and disease outcome. Through the period from 1994-2004, the incidence of meningococcal meningitis ranged from 0.04-1.65 per 100,000 population with the highest incidence recorded for the year 2000, while the lowest incidence was for the year 2004. Serogroup W135 was the most frequent [35.70%], followed by nongroupable [32.90] and serogroup A [17.50%] while the lowest frequency were serogroups X and Y [0.2 /100.000 and 0.06/100.000 population, respectively Makkah region recorded the highest frequency of cases [41%] and Western District [contain Holly places] recorded more than three fourths of cases [76.59%]. All common serogroups were recorded with variation from year to other. Saudi cases represented about one third of cases [30.54%] with the difference was significant compared to Non-Saudi. More than one third of cases were vaccinated among Saudi and Non-Saudi [37.54% and 38.14% respectively]. Males were more than half of cases among Saudi and Non-Saudi. Serogroup W135 was the most frequent among Saudi cases [45.74%], while nongroupable was the most frequent among Non-Saudi cases [45.77%] and the difference was significant. Children <5 years of age recorded higher frequency of cases [29.87%] with serogroup W135 the most frequent among them [52.26%]. Case fatality rate was 31.70% with highest rate among age group 15-29 [45.30%] and those >/= 60 years. the incidence of meningococcal meningitis in this study ranged from 0.04-1.65 per 100,000 populations with the highest incidence recorded for the year 2000, while the lowest incidence was for the year 2004. Serogroup W135 was the most frequent, while the lowest frequenty were serogroups X and Y. Makkah region recorded the highest frequency of cases and western district [contain Holly places] recorded more than three fourths of cases. Case fatality rate generally was 31.70%, with highest rate among age group 15-29 and those >/= 60 years

Enfermedad por meningococo, Neisseria meningitidis: perspectiva epidemiológica, clínica y preventiva / Meningococcal disease caused by Neisseria meningitidis: epidemiological, clinical, and preventive perspectives

La meningitis bacteriana continúa siendo uno de los grandes problemas de la salud pública mundial. En particular, la infección por Neisseria meningitidis afecta tanto a países desarrollados como subdesarrollados, y se presenta en formas endémicas y epidémicas. La enfermedad meningocóccica se puede manifestar clínicamente no sólo como meningitis, sino con cuadros fulminantes de meningococcemia. La persistencia de N. meningitidis se debe al gran porcentaje de portadores y a la dinámica de transmisión de la bacteria. Aproximadamente 500 millones de personas en el mundo son portadoras de N. meningitidis en la nasofaringe. Los factores de transmisiblidad identificados han sido el tabaquismo activo o pasivo, la presencia de infecciones virales del tracto respiratorio superior, épocas de sequía, y el hacinamiento. Por lo anterior, se han descrito brotes de enfermedad meningocóccica en cuarteles militares, escuelas, cárceles y dormitorios. Algunos determinantes que permiten la invasión sistémica incluyen daños en la mucosa nasofaríngea de portadores, cepas virulentas con formación de cápsula, ausencia de anticuerpos bactericidas y deficiencias del sistema del complemento. El control de la enfermedad meningocóccica en circunstancias endémicas y epidémicas se logra por el tratamiento de casos con antibióticos adecuados (penicilina, ceftriaxona o cloranfenicol), la quimioprofilaxis de contactos cercanos (ciprofloxacina, rifampicina o ceftriaxona), y la vigilancia clínica de éstos. Sin embargo, es fundamental subrayar que la clave para el control efectivo de la enfermedad meningocóccica es la inmunoprofilaxis. Las vacunas disponibles incluyen las de polisacáridos monovalentes o bivalentes (serogrupos A y C), tetravalentes (A, C, Y, W-135), la conjugada (C) o la combinada de proteínas de membrana celular y polisacárido (B y C). Recientemente nos hemos visto forzados a establecer planes nacionales de respuesta que incluyen la disponibilidad de vacuna y de medicamentos para quimioprofilaxis, debido a que se ha documentado un incremento de casos de enfermedad por N. meningitidis, serogrupo C, en el país. Es por lo anterior que esta revisión está dirigida a proporcionar al personal de salud un recordatorio de los aspectos relevantes de la epidemiología, y de los clínicos y preventivos de la enfermedad meningocóccica.

Portadores de Neisseria meningitidis en niños de una escuela primaria

Se realizó, con la autorización de la Dirección Municipal de Educación, Dirección Municipal de Salud y el consentimiento informado de los padres, un estudio transversal descriptivo en 318 niños de la Escuela "Mártires del Corynthia"; con el propósito de conocer la prevalencia de portadores de meningococo en niños de edad escolar, determinar los marcadores epidemiológicos de las cepas aisladas y establecer la posible relación existente entre el portador y las variables como edad, sexo, antecedente de infección respiratoria aguda, hacinamiento, amigdalectomía, efecto inhibitorio de la flora acompañante y el estado secretor de antígenos ABH en la saliva. A todos, se les tomó exudado nasofaríngeo y una muestra de saliva. Además, los padres llenaron una encuesta donde se indagó sobre los factores de riesgo a investigar. Se detectó 6,9 % de portadores de meningococo y predominaron las cepas NA:NT:P1.NST:L3,7,9. Los factores de riesgo que dieron resultados estadísticamente significativos respecto a la condición de portador de Neisseria meningitidis fueron: edad, antecedente de infección respiratoria aguda y la presencia de Streptococcus pneumoniae y Neisseria lactamica de la flora bacteriana acompañante en la nasofaringe de los niños investigados.

A cross-sectional and descriptive study was conducted among 318 children from the "Mártires del Corynthia" Primary School under the authorization of the Municipal Division of Education and the informed consent of their parents aimed at knowing the prevalence of meningoccoco carriers in school children, determining the epidemiological markers of the isolated strains and establishing the possible relation existing between the carrier and variables, such as age, sex, acute respiratory infection history, hacinamiento, amigdalectomy, inhibitory effect of of the accompanying flora and the secretory state of ABH antigens in saliva. All of them underwent nasopharyngeal exudate and a saliva sample was taken. In adition, the paents were surveyed about the risks factors to be investigated. 6.9 % of meningoccoco carriers were found and the NA:NT:P1:NST:L3,7,9 strains predominated. The risk factors with statistically significant results regarding the condition of carrier Neisseria meningitidis carrier were age, acute respiratory infection history, and the presence of Streptococcus pneumoniae and Neisseria lactamica of the accompanying bacterial flora in the nasopharynx of the children under study.

Detection of Neisseria meningitidis in cerebrospinal fluid samples from suspicious cases of meningococcal meningitis using polymerase chain reaction and counterimmunoelectrophoresis

Rapid diagnosis of meningococcal disease followed by an early treatment is essential. However, blood or cerebrospinal fluid cultures may not be successful because antibiotic treatment is often started before proper specimens are collected and because bacteria may die during transportation to the laboratory. Improvements in antibiotic therapy for specific microorganisms will require the use of more than one method for immunodiagnosis. In this study a collection of cerebrospinal fluid samples from Brazilian patients was analyzed. Gram stains, culture, counterimmunoelectrophoresis and clinical evaluations for meningococcal diseases were available. The sensitivity of nested PCR (nPCR) was 73 for cerebrospinal fluid of clinically suspected cases, whereas both sensitivity and specificity were 100 when subtypes of Brazilian epidemic strains (P1.7, P1.9 and P1.15) isolated from the samples were used.

Aspectos dos líquidos cefalorraquidianos nas meningites bacterianas / The cerebrospinal fluid in bacterial meningitis

Samples of 1815 cerebrospinal fluid (CSF) were studied in a meningitis outbreak during 1989 in S ao Paulo, Brazil. Neisseria meningitis 56 per cent with 44 per cent type B, Haemophilus influenzae 17 per cent , from which 72 per cent in children (days to 3-year-old) and Streptococcus pneumoniae 14 per cent from which 60 per cent in children (day to 1-year-old) of 443 (24 per cent ) of all strains. Cytochemistry study showed: purulent or turbidity aspects in 70 to 79 per cent positive bacterioscopy or culture of CSF; white cells count > 500/mm3; glucose < 45 mg/dl; protein > 90 mg/dl in 90 per cent of all patients. We concluded that: CSF prognostic factors: (aspect and cytochemistry) were correlated with bacterial meningitis. Bacterioscopy and positive cultures were correlated to NM, SP and HI isolation from these patients (Goodman Test)

Bacteriological study of meningococcal meningitis.

One hundred and thirty samples of cerebro spinal fluid were collected from patients admitted with suspected signs and symptoms of meningococcal meningitis (M. meningitis) during the period from January 1986 to April 1989 and were processed for gram's staining, cultivation and latex agglutination tests for detection of polysaccharide antigen in the CSF. Totally 41.5% of turbid and hazy spinal fluid were positive for N. meningitidis by smear examination. Only 24.6% were positive by culture but 61.5% of sample were positive by latex agglutination tests. All the strains were sensitive to all antibiotics except one strain which was resistant to penicillin but it was sensitive to rifampicin.

A bacteriological study of purulent meningitis in children.

There was an increase in incidence of meningitis in children within the last three years at the Bai Yamunabai Laxman Rao Nair Charitable Hospital in Bombay. Out of 270 purulent samples, 60 (22.22 percent) were culture positive. Neisseria meningitidis showed an increase from one case in 1985 to 11 cases by the end of 1987. Staphylococcus aureus was isolated from 6 percent cases, Streptococcus pneumoniae 15 percent, and Gram negative bacilli were predominant, isolation rate being 57 percent. Salmonella species were isolated from 6 (10 percent) cases. They were multidrug resistant type, some belonging to the rare zoonotic species.