Throat Samples Research Articles

One-year experience with modified BD GeneOhm™ MRSA assay for detection of methicillin-resistant Staphylococcus aureus from pooled nasal, skin, and throat samples

We report our 1-year experience with modified GeneOhm™ MRSA assay (formerly IDI-MRSA) for pooled surveillance specimens in low methicillin-resistant Staphylococcus aureus (MRSA) prevalence clinical setting. We have successfully modified the GeneOhm™ MRSA assay protocol during the specimen preparation step by adding an extra washing step followed by pooling of up to 3 samples per patient (nose, skin, with or without throat) at the lysis step. The sensitivity of the modified assay compared with conventional cultivation was 94.3%, specificity 99.2%, negative predictive value 99.2%, and positive predictive value 94.3%. The modified test is reliable and performed well compared with conventional culture methods in our clinical setting with low-level prevalence of MRSA colonization. Our findings support the use of pooling of the patients samples as a cost-effective way of screening for MRSA colonization.

Sexually transmitted infections among brothel-based sex workers in Tel-Aviv area, Israel: high prevalence of pharyngeal gonorrhoea

Sex workers play a major role in spreading sexually transmitted infections (STIs). We studied the prevalence rates and risk factors for STIs among 300 brothel-based sex workers in Tel-Aviv. Throat swabs were cultured for Neisseria gonorrhoeae, urine samples were tested by polymerase chain reaction (PCR) for Chlamydia trachomatis and N. gonorrhoeae, and sera were tested for syphilis, human immunodeficiency virus (HIV) and type 2 herpes simplex virus (HSV) antibodies. N. gonorrhoeae was cultured from throat samples of 9.0% of participants; PCR testing of urine was positive for C. trachomatis in 6.3% and for N. gonorrhoeae in 5.0%. Syphilis serology was positive (Venereal Disease Research Laboratory [VDRL] titres > 1:8) in 1.3% of women, HSV-2-specific immunoglobulin G was detected in 60% and HIV serology was positive in a single case (0.3%). Having STI was significantly associated with age, number of years in Israel, number of clients a week and condom use for vaginal sex. In a multivariate analysis, having STI was significantly associated with number of clients per week and condom use for vaginal sex. The high prevalence of pharyngeal gonorrhoea reflects most probably the expanding demand of clients for oral sex and the insufficient condom use in this form of sex.

Comparative analysis of emmtype pattern of Group A Streptococcus throat and skin isolates from India and their association with closely related SIC, a streptococcal virulence factor

BackgroundGroup A streptococcus (GAS) causes a wide variety of life threatening diseases in humans and the incidence of such infections is high in developing countries like India. Although distribution of emm types of GAS in India has been described, there is a lack of data describing either the comparative distribution of emm types in throat versus skin isolates, or the distribution of certain virulence factors amongst these isolates. Therefore in the present study we have monitored the emm type pattern of Group A streptococcus throat and skin isolates from India. Additionally, the association of these isolates with closely related sic (crs), a multifunctional compliment binding virulence factor, was also explored.ResultsOf the 94 (46 throat and 48 skin) isolates analyzed, 37 emm types were identified. The most frequently observed emm types were emm49 (8.5%) and emm112 (7.5%) followed by 6.5% each of emm1-2, emm75, emm77, and emm81. Out of 37 emm types, 27 have been previously reported and rest were isolated for the first time in the Indian Community. The predominant emm types of throat (emm49 and emm75) samples were different from those of skin (emm44, emm81 and emm112) samples. After screening all the 94 isolates, the crs gene was found in six emm1-2 (crs1-2) isolates, which was confirmed by DNA sequencing and expression analysis. Despite the polymorphic nature of crs, no intravariation was observed within crs1-2. However, insertions and deletions of highly variable sizes were noticed in comparison to CRS isolated from other emm types (emm1.0, emm57). CRS1-2 showed maximum homology with CRS57, but the genomic location of crs1-2 was found to be the same as that of sic1.0. Further, among crs positive isolates, speA was only present in skin samples thus suggesting possible role of speA in tissue tropism.ConclusionDespite the diversity in emm type pattern of throat and skin isolates, no significant association between emm type and source of isolation was observed. The finding that the crs gene is highly conserved even in two different variants of emm1-2 GAS (speA +ve and -ve) suggests a single allele of crs may be prevalent in the highly diverse throat and skin isolates of GAS in India.

Comparative Analysis of Human Gut Microbiota by Barcoded Pyrosequencing

Humans host complex microbial communities believed to contribute to health maintenance and, when in imbalance, to the development of diseases. Determining the microbial composition in patients and healthy controls may thus provide novel therapeutic targets. For this purpose, high-throughput, cost-effective methods for microbiota characterization are needed. We have employed 454-pyrosequencing of a hyper-variable region of the 16S rRNA gene in combination with sample-specific barcode sequences which enables parallel in-depth analysis of hundreds of samples with limited sample processing. In silico modeling demonstrated that the method correctly describes microbial communities down to phylotypes below the genus level. Here we applied the technique to analyze microbial communities in throat, stomach and fecal samples. Our results demonstrate the applicability of barcoded pyrosequencing as a high-throughput method for comparative microbial ecology.

Clinical validation of a new real-time PCR assay for detection of enteroviruses and parechoviruses, and implications for diagnostic procedures

BackgroundEnteroviruses (EV) and parechoviruses (HPeV) are the most common causes of aseptic meningitis, encephalitis and sepsis-like syndrome in neonates. Detection by nucleic acid amplification methods improves patient management. ObjectiveDevelopment of a real-time PCR assay on a LightCycler for simultaneous detection of EV, HPeV and an internal control to monitor inhibition. Study designWe investigated the value of the new assay, prospectively, in a variety of samples from patients suspected of having viral meningitis or sepsis-like syndrome. ResultsThe assay detected 64 EV serotypes and HPeV types 1–4. Of 186 patients, 63 (33.9%) were EV positive and 18 (9.7%) HPeV positive in one or more samples. In 43 of 159 feces and 6 of 57 throat samples viral culture and PCR were positive. With real-time PCR 27 extra EV and 19 HPeV positives were found.Blood and CSF were present from 33 patients. In 19 patients blood and CSF were positive, one was only positive in CSF, two were only positive in blood, 11 were negative.From 96 patients CSF and/or blood samples were tested and compared to results in throat and/or feces samples. Forty patients were EV-PCR and 14 HPeV-PCR positive in blood and/or CSF. All of these were confirmed by a positive PCR for the respective virus in feces and/or throat. ConclusionsSimultaneous detection of EV and HPeV with this two-step real-time PCR is specific, faster and more sensitive than viral culture. All systemic infections (blood or CSF positive) were confirmed in feces. Culture is no longer necessary for clinical diagnosis and should only be performed on PCR-positive samples to obtain isolates for typing purposes. Application of this assay is an important improvement for patient management since the outcome of the analysis is available within the time frame of clinical decision-making.

Laboratory Approaches to the Diagnosis of Adenovirus Infection Depending on Clinical Manifestations

While commercial enzyme immunoassays (EIA) intended for the detection of adenovirus in fecal specimens are widely used, there are no rapid, convenient, and sensitive commercial tests available for the detection of adenoviruses in respiratory and conjunctival specimens. The applicability of EIA for the detection of adenovirus in stool and throat samples was investigated. One-day rapid culture assay (RCA) for the detection of adenovirus in respiratory and conjunctival specimens was developed and evaluated. Stool samples from patients with gastroenteritis were tested by adenovirus EIA and by cell culture using human embryonic lung cells (HEL) and Graham 293 cells. Blood and stool samples from two BMT patients were also tested for adenovirus by PCR for at least 6 months. Throat specimens from patients with respiratory infections and conjunctival specimens were used for the evaluation of 1-day RCA compared with conventional adenovirus isolation in Graham 293 cells. A total of 3,860 stool samples were tested by EIA Ridascreen, 8,169 by Novitec, and 2,218 by ProSpectT yielding 135 (3.5%), 308 (3.7%), and 77 (3.5%) positive results, respectively. From 305 Ridascreen- and 340 Novitec-negative stool samples, adenoviruses were isolated in three (0.9%) and eight (2.4%) cases, respectively, including two patients undergoing BMT. Multiple sequential stool samples from one BMT patient were repeatedly negative by EIA, but positive by PCR and cell culture. Graham 293 cells were better suited for isolation of adenovirus than HEL. EIA proved unreliable for detecting adenovirus in throat swabs. The sensitivity and specificity of RCA in throat swabs were 90% (37/41) and 100% (64/64), respectively, and 76% (16/21) and 100% (132/132) in conjunctival specimens, respectively. Generally, EIA is sufficiently sensitive for the diagnosis of adenovirus-associated diarrhea. However, it may not be sensitive enough to detect adenovirus in immunocompromised patients undergoing BMT and shedding very few viral particles in stools. Thus, in such cases, a more sensitive assay, such as PCR, is recommended. Furthermore, EIA is not sufficiently sensitive for the reliable detection of adenoviruses in throat swabs. One-day RCA may be useful for the detection of adenoviruses in respiratory and conjunctival specimens.

Vigilância de oito vírus respiratórios em amostras clínicas de pacientes pediátricos no sudeste do Brasil

OBJETIVO: Detecção de oito vírus respiratórios mais comuns: vírus respiratório sincicial humano (VRSH), vírus influenza tipo A e B (IA e IB), vírus da parainfluenza 1, 2 e 3 (VPIH1, 2 e 3), adenovírus (Ad) e metapneumovírus humano (MPVH), a fim de estabelecer a etiologia das infecções respiratórias agudas (IRA) e a epidemiologia desses vírus em crianças pequenas atendidas no Hospital Universitário da Universidade de São Paulo, em São Paulo, Brasil, durante o ano de 2003. MÉTODOS: A vigilância epidemiológica foi realizada em todas as crianças menores de 5 anos hospitalizadas por causa de doenças do trato respiratório inferior (DTRI) entre 1º de janeiro de 2003 e 20 de dezembro de 2003, no hospital universitário. Amostras coletadas de nasofaringe foram analisadas quanto à presença de vírus respiratórios através da reação em cadeia da polimerase e detectadas pelo programa GeneScan. RESULTADOS: Das 336 amostras coletadas, 187 (55,6%) foram positivas para pelo menos um dos vírus respiratórios estudados. De todas as crianças, o VRSH foi identificado em 24,1%, o MPVH em 17,8%, o VPIH3 em 8,3%, o Ad em 6,8%, o IA em 5%, o VPIH1 em 0,6%, sendo que nenhum vírus foi detectado em 44,1%. Infecções virais duplas foram detectadas em 7,1% de todas as amostras (12,8% das amostras positivas). O VPIH2 e o IB não foram detectados no presente estudo. CONCLUSÕES: Este estudo confirma que as crianças menores de 5 anos, e especialmente aquelas menores de 1 ano, apresentam uma alta taxa de hospitalização devido aos seguintes vírus: VRSH, MPVH, VPIH, influenza e adenovírus. Foi possível determinar a etiologia e epidemiologia da maioria das IRAs e traçar o perfil de sazonalidade dos vírus respiratórios mais comuns entre as crianças pequenas.

Surveillance of eight respiratory viruses in clinical samples of pediatric patients in Southeast Brazil

Detection of the eight most common respiratory viruses: human respiratory syncytial virus (HRSV), influenza virus A and B (IA and IB), parainfluenza viruses 1, 2 and 3 (HPIV1, 2 and 3), adenovirus (Ad) and human metapneumovirus (HMPV), in order to establish the etiology of acute respiratory infections (ARIs) and the epidemiology of these viruses in young children seen at Hospital Universitário, Universidade de São Paulo, in São Paulo, Brazil, during 2003. The epidemiological surveillance was conducted in all children younger than 5 years hospitalized at the Hospital for lower respiratory tract infections (LRTI) from January 1, 2003 to December 30, 2003. Nasal and throat samples were scanned for respiratory viruses by polymerase chain reaction and detected by the GeneScan assay. Of 336 samples collected from 336 patients, 187 (55.6%) were positive for at least one of the respiratory viruses studied. Of all the children, HRSV was identified in 24.1%, HMPV in 17.8%, HPIV3 in 8.3%, Ad in 6.8%, IA in 5%, HPIV1 in 0.6%, but no virus could be detected in 44.1%. Dual virus infections were detected in 7.1% of all samples (12.8% of positive samples). HPIV2 and IB were not detected in the present study. This study confirms that children younger than 5 years and particularly younger than 1 year have a high hospitalization rate due to HRSV, HMPV, HPIV, influenza and adenovirus. We were able to determine the etiology and epidemiology of most ARIs and trace the seasonal profile of the commonest respiratory viruses among young children.

Molecular and phylogenetic analysis of human influenza virus among Iranian patients in Shiraz, Iran

Influenza is a viral respiratory pathogen responsible for frequent seasonal epidemics. There are currently three major human influenza viruses in global circulation namely A/H1N1, A/H3N2, and B. The objective of this study was to determine the human influenza virus genotypes in Shiraz, the capital of the Fars province of Iran. Three hundred patients suspected with human influenza virus infection were enrolled in this survey (2004-2005). The throat samples were cultured and titrated by hemagglutination (HA) assay. Typing and subtyping were performed by an in-house developed multiplex RT-PCR. Moreover, the phylogenetic analysis was carried out for HA gene. A total of 24 samples were found to be positive for human influenza virus infection, 17 H1N1 and 7 H3N2. These results were in agreement with the HI assay. The phylogenetic analysis results revealed that the Iranian H1N1 isolated were close to the A/New Caledonia/20/99 vaccine strain genetically and the Iranian H3N2 isolates were also related closely to the Fujian/411/021 and California/7/2004 vaccine strains. However, a slight genetic drift was found in these isolates. In conclusion, it was demonstrated that both influenza A subtypes A/H1N1 and A/H3N2 were dominant among Iranian patients in Shiraz during the 2004/5 winter season.

Significance of oral Candida infections in children with cancer

Candidiasis is common in children with cancer, particularly during periods of severe immunosuppression and neutropenia. Our aim was to study the microbiological changes in the oral cavity of children with newly diagnosed cancer. The study group consisted of 30 consecutive children and adolescents, 16 with acute lymphoblastic leukemia and 14 with solid tumors. Oral cultures to detect fungi and bacteria were conducted for all patients before treatment, during and after neutropenic episodes. In 23 patients developing fever simultaneous throat, urine and blood sampling was carried out. No pathogens were found in the cultures taken before the outset (30 cultures) or after recovery from (30 cultures) the neutropenic episodes. In the 45 oral cultures taken during the neutropenic episodes 38 (84.4%) proved positive. Fungi were the most frequently isolated oral pathogens: 33/38 yeast and 6/38 bacterial infections were identified. There was no association between the underlying malignancy and the occurrence of the positive cultures. Of the 30 patients, all 23 (76.7%) who have developed moderate-to-severe neutropenia, developed oral fungal colonization or clinically obvious fungal infection at least on one occasion during the study. In addition to oral samples, fungi were identified in 9/23 pharyngeal swabs, 6/23 urine and 1/23 blood cultures. The initial fungal pathogen was exclusively (33/33) Candida albicans. In extended severe neutropenic states, C. albicans was replaced by non-albicans species (C. kefyr, C. lusitaniae, C. sake, C. tropicalis) in 5 patients between 4 to 6 days of the neutropenic episodes. Four of the nonalbicans Candida strains were resistant to azole-type antifungal agents. Neutropenic episodes of children with cancer are associated with an increased risk of developing oral and even systemic infections with C. albicans that can be replaced by azole-resistant nonalbicans strains in prolonged neutropenia contributing to morbidity of these patients.

Prevalence and mechanisms of macrolide resistance inStreptococcus pyogenesin Ankara, Turkey

From 2002 to 2003, a total of 381 consecutive S. pyogenes isolates were obtained from throat swabs (n = 337) and samples of pus (n = 31), sputum (n=10) and blood (n = 3) at Hacettepe University Hospital. The susceptibility of the isolates to erythromycin was tested by the agar dilution method. Erythromycin resistant strains were then tested for their MICs to azithromycin, clindamycin, and penicillin, their phenotype of resistance to macrolides-lincosamides-streptogramin B (MLSB) and for the presence of macrolide resistance genes. The rate of resistance to erythromycin was 6.8%. Constitutive (cMLSB), inducible (iMLSB), and M phenotypes of resistance were detected in 7.7, 30.8, and 57.7% of resistant strains, respectively. One strain had both cMLSB and iMLSB phenotypes. All M phenotypes carried the mefA gene, all iMLSB phenotype carried the ermTR gene, 1 isolate with cMLSB phenotype harboured the ermB gene, and 1 isolate with cMLSB phenotype carried both the ermB and mefA genes. One strain which showed cMLSB and iMLSB phenotypes harboured the ermB gene.

No evidence of Legionella infection in general practice patients presenting with acute respiratory infections in The Netherlands

The role of Legionella spp. in the aetiology of acute respiratory infections (ARIs) is largely unknown. In this case‐control study, conducted in a general practitioner setting during 2000 and 2001, nose and throat samples from patients presenting with ARIs (n = 230) and controls (n = 200) were analysed for the presence of Legionella spp. by real‐time PCR. Legionella DNA was not detected in any of the cases or controls. Thus, Legionella spp. do not seem to play a role in patients presenting with ARIs, nor were they present in patients who visited their general practitioner for complaints other than ARIs.

Effect of Mupirocin Treatment on Nasal, Pharyngeal, and Perineal Carriage of Staphylococcus aureus in Healthy Adults

Nasal carriage of Staphylococcus aureus is an important risk factor for S. aureus infections. Mupirocin nasal ointment is presently the treatment of choice for decolonizing the anterior nares. However, recent clinical trials show limited benefit from mupirocin prophylaxis in preventing nosocomial S. aureus infections, probably due to (re)colonization from extranasal carriage sites. Therefore, we studied the effectiveness of mupirocin nasal ointment treatment on the dynamics of S. aureus nasal and extranasal carriage. Twenty noncarriers, 26 intermittent carriers, and 16 persistent carriers had nasal, throat, and perineum samples taken 1 day before and 5 weeks after mupirocin treatment (twice daily for 5 days) and assessed for growth of S. aureus. The identities of cultured strains were assessed by restriction fragment length polymorphisms of the coagulase and protein A genes. The overall carriage rate (either nasal, pharyngeal, or perineal carrier or a combination) was significantly reduced after mupirocin treatment from 30 to 17 carriers (P = 0.003). Of the 17 carriers, 10 (60%) were still colonized with their old strain, 6 (35%) were colonized with an exogenous strain, and 1 (5%) was colonized with both. Two noncarriers became carriers after treatment. The acquisition of exogenous strains after mupirocin treatment is a common phenomenon. The finding warrants the use of mupirocin only in proven carriers for decolonization purposes. Mupirocin is effective overall in decolonizing nasal carriers but less effective in decolonizing extranasal sites.

An outbreak of food-borne group A Streptococcus (GAS) tonsillopharyngitis among residents of a dormitory

Epidemics of food-borne pharyngitis due to group A Streptococcus are rarely reported. Here we present an outbreak of food-borne tonsillopharyngitis in female dormitories in the Islamic Republic of Iran. Throat swabs and cultures were performed on a number of patients, and of specimens from the nasopharynx and hands of staff who were involved in food processing. We planned a case-control study for assessing the source of epidemics. 11 out of 17 throat swabs of students were positive for Streptococcus group A and also 2 throat samples from asymptomatic cooks were positive. A DNA fingerprinting study showed that Streptococcus group A strains of 11 students and 1 cook had the same T agglutination pattern and M protein factor (M3/T13). It is suggested that group A streptococci as well as group C and G streptococci can cause epidemic food-borne pharyngitis. Regular health surveillance of food handlers and food preparation processes are important for prevention of such outbreaks.

Population-based surveillance for hospitalizations associated with respiratory syncytial virus, influenza virus, and parainfluenza viruses among young children.

Respiratory syncytial virus (RSV), influenza virus, and parainfluenza viruses (PIV) cause significant morbidity in young children. Although only influenza virus infection and illness is currently vaccine-preventable, vaccines are under development for RSV and PIV. We established a prospective, active population-based surveillance network to provide precise estimates of hospitalization rates for viral acute respiratory illness (ARI) in young children and to measure the potential impact of enhanced vaccine usage on these rates. Prospective, active population-based surveillance was conducted in young children who were hospitalized for ARI from October 1, 2000, to September 30, 2001, in Monroe County, New York (Rochester area) and Davidson County, Tennessee (Nashville area). Eligible children younger than 5 years were those who resided in surveillance counties and were hospitalized for febrile or acute respiratory illness. Viral culture and polymerase chain reaction identified viruses from nasal and throat samples obtained from all surveillance children. We measured population-based rates of hospitalization for RSV, influenza virus, and PIV as well as demographic, clinical, and risk factor assessment for each virus. Of 812 eligible hospital admissions, 592 (73%) children were enrolled. Of the enrolled children, RSV was identified in 20%, influenza in 3%, PIV in 7%, other respiratory viruses in 36%, and no detectable virus in 39%. Population-based rates of ARI hospitalizations in children younger than 5 years were 18 per 1000. Virus-positive hospitalization rates per 1000 children were 3.5 for RSV, 1.2 for PIV, and 0.6 for influenza virus. Younger age (particularly <1 year), black and Hispanic race/ethnicity, male gender, and presence of chronic underlying illness were associated with higher hospitalization rates. This study confirms that children younger than 5 years and particularly children younger than 1 year have a high burden of hospitalization from RSV, influenza, and PIV. The enhanced use of influenza vaccine and the development of RSV and PIV vaccines have the potential to reduce markedly the pediatric morbidity from ARIs.

Detection of aerosolized varicella-zoster virus DNA in patients with localized herpes zoster.

We examined the excretion of varicella zoster virus (VZV) in hospitalized patients with herpes zoster localized to the thoracic region whose skin lesions were covered with either hydrocolloid dressing agents (hydrocolloid group) or conventional gauze bandages (gauze group). The presence of VZV DNA in swab samples from lesion coverings, the throat, and filters of air purifiers was examined by use of a sensitive polymerase chain reaction assay. For the hydrocolloid group, VZV was detected in none of the samples from lesion coverings or air purifier filters; for the gauze group, VZV DNA was detected in samples from gauze coverings and air purifier filters for all 6 patients. VZV DNA was detected less frequently in throat samples from patients in the hydrocolloid group than in those from patients in the gauze group. The results of the present study suggest that hydrocolloid dressing agents prevent excretion of aerosolized VZV DNA from skin lesions of patients with localized herpes zoster.

Oropharyngeal carriage of macrolide-resistant viridans group streptococci: a prevalence study among healthy adults in Belgium.

Viridans group streptococci (VGS) are gaining significance as reservoirs of resistance determinants for respiratory tract pathogens. Our aim was to investigate healthy adults for oropharyngeal carriage of VGS that are resistant to macrolides, as well as to other common antibiotics. Macrolide-resistant VGS were isolated from throat samples of 154 healthy Belgian adults, and phenotyped and genotyped for erm(A), erm(B) and mef(A). In vitro susceptibilities to 10 antimicrobials and the presence of tetracycline resistance genes were also determined. Carriage was detected in 71% of the population screened, from whom 157 unique, macrolide-resistant VGS were isolated. A constitutive (cMLS) phenotype was present in 105 isolates, of which 102 isolates carried either erm(B) or erm(B) + mef(A). The remaining three isolates did not present with any of the genes studied. All 45 isolates showing the M phenotype carried mef(A). The least abundant inducible (iMLS) isolates (n = 7) carried either erm(B) or erm(B) + mef(A). The most abundant macrolide-resistant VGS species was Streptococcus mitis (51%). Co-resistance to tetracycline was identified in 114 isolates, of which tet(M) was present in 105, tet(O) in two and both tet(M) and tet(O) in one, while the remaining six isolates did not present with either gene. tet(M) was also present in four tetracycline susceptible and two intermediately resistant isolates. Fluoroquinolone resistance (ciprofloxacin MIC > or = 4 mg/L) was detected in 16 isolates. Resistance to telithromycin, penicillin and chloramphenicol was appreciably low. This study highlights a high oropharyngeal carriage of macrolide-resistant VGS and its co-resistance to tetracycline and fluoroquinolones among healthy Belgian adults.

Etiologic Diagnosis of 204 Pericardial Effusions

The etiologic evaluation of pericardial effusion is frequently unsuccessful when noninvasive methods are used. To determine the cause of the current episode, all patients with echographically identified pericardial effusion from May 1998 to December 2002 underwent noninvasive diagnostic testing of blood, throat, and stool samples. Patients with postpericardiotomy syndrome were excluded. To analyze the value of our tests, we tested randomly selected blood donors as negative controls. Among 204 included patients, 107 (52.4%) had a final etiologic diagnosis: the etiology of 52 was highly suspected at first examination and later confirmed (thyroid deficiency, 5 cases; systemic lupus erythematous, 7; rheumatoid arthritis, 7; scleroderma, 3; cancer, 25; and renal insufficiency, 5). A definite etiologic diagnosis was made in 11 patients from pericardial fluid analysis (cancer, 5 cases; tuberculosis, 3; Streptococcus pneumoniae, Citrobacter freundii, and Actinomyces, 1 case each). Among 141 patients considered to have idiopathic pericarditis, 44 (32.1%) gained an etiologic diagnosis by our systematic testing strategy. This included serologic evaluation of serum (Coxiella burnetii, 10 cases; Bartonella quintana, 1; Legionella pneumophila, 1; Mycoplasma pneumoniae, 4; influenza virus, 1), viral culture of throat swabs (enterovirus, 8 cases; and adenovirus, 1), high-level antinuclear antibodies (>1/400, 3 cases), and thyroid-stimulating hormone (15 abnormal results). Antibodies to Toxoplasma and cytomegalovirus, enterovirus recovered from rectal swabs, and low-level antinuclear antibodies were seen with equal frequency in patients and controls. Using our evaluation strategy, the number of pericardial effusions classified as idiopathic was less than in other series. Systematic testing for Q fever, Mycoplasma pneumoniae, thyroid abnormalities, and antinuclear antibodies, accompanied by viral throat cultures, frequently enabled us to diagnose diseases not initially suspected in patients with pericardial effusion.

Inhalation efficacy of RFI-641 in an African green monkey model of RSV infection.

Human respiratory syncytial virus (RSV) is a major cause of acute upper and lower respiratory tract infections. RFI-641 is a novel RSV fusion inhibitor with potent in vitro activity. In vivo efficacy of RFI was determined in an African green monkey model of RSV infection involving prophylactic and therapeutic administration by inhalation exposure. Inhalation was with an RFI-641 nebulizer reservoir concentration of 15 mg/ml for 15 minutes (short exposure) or 2 hours (long exposure). Efficacy and RFI-641 exposure was determined by collection of throat swabs, nasal washes and bronchial alveolar lavage (BAL) on selected days. The short-exposure group (15 minutes) exhibited no effect on the nasal, throat or BAL samples. The throat and nasal samples for the long-exposure group failed to show a consistent reduction in viral titers. RFI-641 2 hours exposure-treated monkeys showed a statistically significantly log reduction for BAL samples of 0.73-1.34 PFU/ml (P-value 0.003) over all the sampling days. Analysis indicates that the long-exposure group titer was lower than the control titer on day 7 and when averaged across days. The results of this study demonstrate the ability of RFI-641 to reduce the viral load of RSV after inhalation exposure in the primate model of respiratory infection.

Three ECHOvirus serotypes responsible for outbreak of aseptic meningitis in Rhône-Alpes region, France.

During the year 2000 in the Rhône-Alpes region of France, 559 cases of aseptic meningitis due to enterovirus infection were recorded (approximate incidence, 10 cases per 100,000 population compared with a mean of 0.3 cases during endemic years in this region; P<0.001). Cerebrospinal fluid samples were collected from all 559 patients and processed for enterovirus RNA detection using a reverse transcriptase polymerase chain reaction assay only. In addition to the cerebrospinal fluid samples, 40 stool and 76 throat samples were collected from 116 patients (20.7% of cases); the following three enterovirus serotypes were isolated from the stool and throat samples only: ECHOvirus 13 (48/116; 41.3%), ECHOvirus 30 (44/116;37.9%) and ECHOvirus 6 (24/116, 20.7%). This is the first report that demonstrates the involvement of three different ECHOvirus serotypes, particularly ECHOvirus 13, in an outbreak of aseptic meningitis, in the French Rhône-Alpes region.