Streptococcus pneumoniae outbreak in a rural Regina community.
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Adolescent↗
Biomedical subjects
P N Levett
Publications and source records attributed to P N Levett.
Evaluation of a commercial latex agglutination assay for serological diagnosis of leptospirosis.
Leptospirosis is a febrile zoonosis of worldwide distribution. A latex agglutination assay was evaluated in two studies, the first using a panel of well-characterized sera from patients with leptospirosis and from patients with other disease states and the second, a prospective hospital-based study, evaluating sera from 186 consecutive patients admitted to hospital with acute febrile illness. The confirmed leptospirosis serum panel included paired acute- and convalescent-phase specimens from 40 cases, of which 34 gave positive latex tests (case sensitivity, 85%; 95% confidence interval [95% CI], 70 to 94%). The other diseases represented in the panel of 112 specimens from nonleptospirosis patients included autoimmune diseases, brucellosis, dengue, melioidosis, malaria, syphilis, toxoplasmosis, viral hepatitis, and a number of other viral infections. The specificity of latex agglutination using this panel was 81% (95% CI, 73 to 87%). Among the patients with acute febrile illness, there were 25 cases of leptospirosis and 161 patients with other diagnoses. The sensitivity and specificity of latex agglutination in this group were 88% (95% CI, 72 to 97%) and 98% (95% CI, 95 to 100%), respectively. In this evaluation, the two distinct groups of specimens gave similar results for sensitivity, but specificity was different in each study. The sensitivity and specificity observed for the hospital study were similar to those obtained in evaluations of other rapid tests in the same population. The results of this study suggest that multiple evaluations of new diagnostic assays should be performed, because performance characteristics may vary in different populations.
Antibodies, Bacterial↗
Seroprevalence of HSV-1 and HSV-2 in Barbados.
Type-specific enzyme-linked immunosorbent assays, based upon recombinant glycoprotein G (gG), were used to detect antibodies to HSV-1 and HSV-2, in a small Caribbean island population. A blinded serosurvey was performed on samples from 184 blood donors, 122 pregnant women, and 120 HIV-positive patients. The seroprevalence of HSV-1 and HSV-2 was 81% and 34%, respectively, in blood donors, 84% and 40% in the antenatal population and 89% and 77% in the HIV-positive group. As expected the majority of adults were seropositive against HSV-1. However, the HSV-2 seroprevalence was significantly higher in HIV-infected adults than in the other groups. These findings support the need for prospective epidemiological studies in this population.
Antibodies, Viral↗
Reclassification of Leptospira parva Hovind-Hougen et al. 1982 as Turneriella parva gen. nov., comb. nov.
Analysis of the G+C content, DNA-DNA relatedness to other leptospires and 16S rRNA gene sequence of Leptospira parva showed that this species was not related to other Leptospira species. On the basis of these data, it is proposed that Leptospira parva should be transferred to the genus Turneriella as Turneriella parva gen. nov., comb. nov., with strain H(T) (=NCTC 11395(T)=ATCC BAA-1111(T)) as the type strain.
Bacterial Typing Techniques↗
Use of immunoglobulin G avidity assays for differentiation of primary from previous infections with West Nile virus.
Since its introduction in 1999, West Nile virus (WNV) infections have spread rapidly across the North American continent. Diagnosis of acute WNV infection by detection of WNV-specific immunoglobulin M (IgM) is complicated by the persistence of detectable IgM for more than 1 year in some patients. IgG antibody avidity testing was assessed as a supplemental assay in the diagnosis of current infections. Three groups of serum samples were assayed in parallel by two different IgG avidity test systems (indirect immunofluorescence test [IIFT] and prototype enzyme-linked immunosorbent assay [ELISA]; EUROIMMUN, Luebeck, Germany). Group I (40 sera taken between 2 and 9 days after the onset of influenza-like symptoms) and group II (40 sera taken between 10 and 43 days after onset) were acute and convalescent specimens from patients with a positive anti-WNV IgM test (ELISA; Focus Diagnostics, Cypress, CA). Group III consisted of 43 patient sera collected between 6 and 12 months after infection. IgG antibodies specific for WNV were detected in 38% (ELISA) and 50% (IIFT) of group I sera, in 90% (ELISA and IIFT) of group II sera, and in 100% (ELISA and IIFT) of group III sera. Low-avidity IgG antibodies were demonstrated in 86% (ELISA) and 95% (IIFT) of IgG-positive patient samples taken between 2 and 43 days after the onset of symptoms (groups I and II). High-avidity IgG antibodies were detected in 100% of group III sera obtained 6 months or more after the onset of symptoms (ELISA and IIFT). IgG avidity tests for WNV infections are rapid and simple to perform. The determination of IgG avidity provides additional diagnostic certainty in differentiating between recently acquired and previous infections with WNV.
Antibodies, Viral↗
Failure to detect Angiostrongylus cantonensis in rats in Barbados.
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Angiostrongylus cantonensis↗
Human external ophthalmomyiasis occurring in Barbados.
Human infection with the sheep nasal botfly Oestrus ovis occurs sporadically. In most cases, there is a history of a strike in the eye by the adult fly. Human O. ovis has been reported rarely from the Americas. We report the first case of O. ovis infection in the Caribbean region, which occurred in an urban area of Barbados. The patient responded to removal of the larvae from the conjunctiva and symptomatic treatment.
Animals↗
Serological evidence of hantavirus in humans and rodents in Barbados.
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Adolescent↗
Leptospiral carriage by mice and mongooses on the island of Barbados.
Leptospirosis is a zoonotic disease, maintained by chronic infection of the kidneys of reservoir animals, usually small mammals. Infection in humans is acquired from direct or indirect exposure to the urine of infected animals. Leptospirosis has a high incidence in tropical regions, and has been studied extensively in several Caribbean countries. We studied the carriage of Leptospira serovars by two small mammals which are potential maintenance hosts of the disease in Barbados. A total of 136 mongooses (Herpestes auropunctatus) and 97 mice (Mus musculus) were caught in live traps. Leptospiral antibodies were detected by microscopic agglutination test (MAT) using antigens representing 12 serogroups, and kidney tissues were inoculated into polysorbate medium for isolation of leptospires. The seroprevalence (at a titre of > or = 100) in mice was 28.2% (24/85, 95% CI 19.0, 39.1) and in mongooses 40.7% (48/118, 95% CI 31.7, 50.1). In mice, antibodies were detected predominantly against serogroups Ballum and Autumnalis, while in mongooses the predominant serogroup was Autumnalis. Leptospires were isolated from 28 mice (28.9%, 95% CI 20.1, 39.0) and from 4 mongooses (2.9%, 95% CI 0.8, 7.4). Mouse isolates were identified as serovars arborea (17) and bim (7). As in other parts of the world, common house mice (Mus musculus) represent a significant reservoir of leptospirosis. Although carriage of the Ballum serovar, arborea, was not unexpected, this represents the first time that an animal reservoir of serovar bim has been identified. This is significant because bim causes about 63% of human leptospirosis in Barbados, and control efforts and education for prevention can now be targeted at a specific reservoir.
Animals↗
Testing for gene-gene interaction controlling total IgE in families from Barbados: evidence of sensitivity regarding linkage heterogeneity among families.
Genetic heterogeneity has been proposed as a hallmark feature of allergic disease. To test the hypothesis that total IgE levels are jointly influenced by a locus on chromosome 12q21.1-q21.31 and a locus on 17q11.2-q21.2, we conducted multipoint allele-sharing analyses using nonparametric linkage (NPL) methods on Afro-Caribbean families from Barbados to test for evidence of gene-gene interactions. Significant correlations were observed between NPL scores at D12S1052 and both D17S1293 and D17S1299 for a dichotomized phenotype of total IgE. An analysis of family-specific NPL scores revealed that evidence for interaction was being driven largely by one multiplex pedigree (NPL = 12.01, 12.23, and 12.16 at D12S1052, D17S1293, and D17S1299, respectively). Using the programs SIMWALK (v2.0) and GOLD, a different set of haplotypes in this influential family was observed around D12S1052 and the 17q loci compared to the other Barbados pedigrees. Our findings are a classic example of founder effect, provide evidence for sensitivity of this type of linkage analysis to unusual pedigrees, and highlight an element of genetic heterogeneity that has been given little attention in the study of complex traits.
Asthma↗
Persistence of anti-leptospiral IgM, IgG and agglutinating antibodies in patients presenting with acute febrile illness in Barbados 1979-1989.
The persistence of anti-leptospiral IgM and IgG antibodies and agglutinating antibodies was studied in serologically confirmed cases of severe leptospirosis during the acute illness and over periods of several years after recovery. The antibody response in non-leptospirosis patients presenting to hospital with similar symptoms over the same period of time was used to estimate the background antibody level to leptospirosis in the community. All patients enrolled in the study had blood samples collected twice in the acute stage of illness, once during convalescence and then annually from the time of initial hospitalisation until the end of the study period. Six hundred and thirty-eight patients presented to hospital with acute febrile illness, of whom 321 were diagnosed with leptospirosis. Patients who had severe leptospirosis commonly remained seropositive, with IgM, IgG and agglutinating antibodies detectable for several years after infection. A significant proportion of cases had high titres of agglutinating antibody detectable by the microscopic agglutination test (> or = 800). There were marked differences in the magnitude and duration of persistence of agglutinating antibodies directed against different serogroups. More than 20% of cases with evidence of infection with serogroup Autumnalis retained titres of >800, 4 years after the acute illness. In one case a titre of 800 was detected 11 years after infection. Persistence of agglutinating antibody titres can create problems in interpretation of serological results and make it impossible to estimate the time of infection, given a specific titre. This study demonstrates that in endemic areas where seroprevalence is high, use of a single elevated titre is not reliable to define a current infection.
Agglutination Tests↗
Two methods for rapid serological diagnosis of acute leptospirosis.
Leptospirosis is a common and underdiagnosed zoonosis. Two rapid assays for serological diagnosis of acute leptospirosis in diagnostic laboratories, the immunoglobulin M (IgM)-dipstick assay and the indirect hemagglutination assay (IHA), were evaluated and compared with standard assays. Sera were examined from 104 patients admitted to a hospital for investigation in a leptospirosis diagnostic protocol. Specimens for serology were taken on days 1 and 4 of the patients' hospital stay. Antibodies were detected using an IgM-enzyme-linked immunosorbent assay (ELISA), microscopic agglutination test (MAT), an IgM-dipstick assay, and an IHA. Fifty-one patients were found to have leptospirosis. The sensitivity of the IgM-dipstick assay was 98%, its specificity was 90.6%, its positive predictive value was 90.9%, and its negative predictive value was 98%. The sensitivity of the IHA was 92.2%, its specificity was 94.4%, its positive predictive value was 95.9%, and its negative predictive value was 92.7%. The standard IgM-ELISA and MAT, were positive in the first samples tested from 67 and 55% of the cases, respectively, and the rapid IgM-dipstick assay and IHA were positive in 71 and 49%, respectively, in the first sample tested. Both rapid assays are highly sensitive and specific. Neither requires specialized equipment, and both are suitable for use in diagnostic laboratories.
Acute Disease↗
Leptospirosis.
Leptospirosis is a worldwide zoonotic infection with a much greater incidence in tropical regions and has now been identified as one of the emerging infectious diseases. The epidemiology of leptospirosis has been modified by changes in animal husbandry, climate, and human behavior. Resurgent interest in leptospirosis has resulted from large outbreaks that have received significant publicity. The development of simpler, rapid assays for diagnosis has been based largely on the recognition that early initiation of antibiotic therapy is important in acute disease but also on the need for assays which can be used more widely. In this review, the complex taxonomy of leptospires, previously based on serology and recently modified by a genotypic classification, is discussed, and the clinical and epidemiological value of molecular diagnosis and typing is also evaluated.
Animals↗
Leptospiral proteins recognized during the humoral immune response to leptospirosis in humans.
Leptospirosis is an emerging zoonosis caused by pathogenic spirochetes belonging to the genus Leptospira. An understanding of leptospiral protein expression regulation is needed to develop new immunoprotective and serodiagnostic strategies. We used the humoral immune response during human leptospirosis as a reporter of protein antigens expressed during infection. Qualitative and quantitative immunoblot analysis was performed using sera from 105 patients from Brazil and Barbados. Sera from patients with other diseases and healthy individuals were evaluated as controls. Seven proteins, p76, p62, p48, p45, p41, p37, and p32, were identified as targets of the humoral response during natural infection. In both acute and convalescent phases of illness, antibodies to lipopolysaccharide were predominantly immunoglobulin M (IgM) while antibodies to proteins were exclusively IgG. Anti-p32 reactivity had the greatest sensitivity and specificity: positive reactions were observed in 37 and 84% of acute- and convalescent-phase sera, respectively, while only 5% of community control individuals demonstrated positive reactions. Six immunodominant antigens were expressed by all pathogenic leptospiral strains tested; only p37 was inconsistently expressed. Two-dimensional immunoblots identified four of the seven infection-associated antigens as being previously characterized proteins: LipL32 (the major outer membrane lipoprotein), LipL41 (a surface-exposed outer membrane lipoprotein), and heat shock proteins GroEL and DnaK. Fractionation studies demonstrated LipL32 and LipL41 reactivity in the outer membrane fraction and GroEL and DnaK in the cytoplasmic fraction, while p37 appeared to be a soluble periplasmic protein. Most of the other immunodominant proteins, including p48 and p45, were localized to the inner membrane. These findings indicate that leptospiral proteins recognized during natural infection are potentially useful for serodiagnosis and may serve as targets for vaccine design.
Animals↗
Antibiotic susceptibility of Clostridium difficile isolates from adult patients at two Jamaican hospitals. Clinical and epidemiological implications.
The susceptibility of 39 toxin producing Clostridium difficile isolates from stools of hospitalized patients was determined, by disc diffusion, to six antibiotics. All but one isolate (toxin A negative) produced toxin A and toxin B. A wide variation in susceptibility to clindamycin, tetracycline and chloramphenicol was noted. Erythromycin and cotrimoxazole showed a clear-cut discrimination in resistance and susceptibility, while all isolates were sensitive to vancomycin. Erythromycin sensitive isolates demonstrated a significant association with diarrhoea (60.9%, 14/23, p < 0.001). These strains were predominantly found at the University Hospital of the West Indies (UHWI, 94.1%, 16/17). Strains resistant to erythromycin and clindamycin together were commonly found at the National Chest Hospital (NCH, 68.2%, 15/22). All erythromycin sensitive strains found at the NCH were from patients transferred to that hospital. These findings suggest that there is a common strain of C difficile (erythromycin resistant) at the NCH different from that found at the UHWI; the resistant pattern seen with isolates from the NCH was typical of toxigenic serogroup C strain and could be typed by the the disc diffusion method. Patients at the NCH who were colonized with either of the two strains of C difficile were likely to get diarrhoea, once there was suppression of the normal microflora by antibiotics and colonic overgrowth with C difficile.
Adolescent↗
Antimicrobial sensitivity of Haemophilus influenzae isolates from bacterial conjunctivitis.
Haemophilus influenzae is one of the common bacterial pathogens which affect children. Resistance to frequently used antibiotics is becoming a significant problem in community isolates of common pathogens. A retrospective review was conducted of the serotypes and antimicrobial sensitivity of H influenzae isolates from bacterial conjunctivitis, over an 18-month period. Data on antimicrobial sensitivity (obtained by the National Committee for Clinical Laboratory Standards disk diffusion method) and beta-lactamase production, and typing results, were analysed. Ninety-nine isolates were recovered, of which 87 were typed. Most isolates were recovered from children under one year of age. Ninety-three percent were unencapsulated and biotypes I and IV were most common. H influenzae type b was recovered only twice. beta-lactamase was produced by 41% isolates while four isolates were ampicillin-resistant but did not produce beta-lactamase. All isolates were sensitive to chloramphenicol and 45% were co-trimoxazole sensitive. H influenzae is commonly isolated from bacterial conjunctivitis in Barbados and, as elsewhere, the majority of isolates are from small children and are non-encapsulated. However, there is a high prevalence of beta-lactamase production, which may serve as a reservoir for transfer to more invasive encapsulated strains of H influenzae within the oropharyngeal flora.
Adult↗
Atopic dermatitis is associated with a functional mutation in the promoter of the C-C chemokine RANTES.
Up-regulation of C-C chemokine expression characterizes allergic inflammation and atopic diseases. A functional mutation in the proximal promoter of the RANTES gene has been identified, which results in a new consensus binding site for the GATA transcription factor family. A higher frequency of this allele was observed in individuals of African descent compared with Caucasian subjects (p < 0.00001). The mutant allele was associated with atopic dermatitis in children of the German Multicenter Allergy Study (MAS-90; p < 0.037), but not with asthma. Transient transfections of the human mast cell line HMC-1 and the T cell line Jurkat with reporter vectors driven by either the mutant or wild-type RANTES promoter showed an up to 8-fold higher constitutive transcriptional activity of the mutant promoter. This is the first report to our knowledge of a functional mutation in a chemokine gene promoter. Our findings suggest that the mutation contributes to the development of atopic dermatitis. Its potential role in other inflammatory and infectious disorders, particularly among individuals of African ancestry, remains to be determined.
Alleles↗
The leptospiral major outer membrane protein LipL32 is a lipoprotein expressed during mammalian infection.
We report the cloning of the gene encoding the 32-kDa lipoprotein, designated LipL32, the most prominent protein in the leptospiral protein profile. We obtained the N-terminal amino acid sequence of a staphylococcal V8 proteolytic-digest fragment to design an oligonucleotide probe. A Lambda-Zap II library containing EcoRI fragments of Leptospira kirschneri DNA was screened, and a 5.0-kb DNA fragment which contained the entire structural lipL32 gene was identified. Several lines of evidence indicate that LipL32 is lipid modified in a manner similar to that of other procaryotic lipoproteins. The deduced amino acid sequence of LipL32 would encode a 272-amino-acid polypeptide with a 19-amino-acid signal peptide, followed by a lipoprotein signal peptidase cleavage site. LipL32 is intrinsically labeled during incubation of L. kirschneri in media containing [(3)H]palmitate. The linkage of palmitate and the amino-terminal cysteine of LipL32 is acid labile. LipL32 is completely solubilized by Triton X-114 extraction of L. kirschneri; phase separation results in partitioning of LipL32 exclusively into the hydrophobic, detergent phase, indicating that it is a component of the leptospiral outer membrane. CaCl(2) (20 mM) must be present during phase separation for recovery of LipL32. LipL32 is expressed not only during cultivation but also during mammalian infection. Immunohistochemistry demonstrated intense LipL32 reactivity with L. kirschneri infecting proximal tubules of hamster kidneys. LipL32 is also a prominent immunogen during human leptospirosis. The sequence and expression of LipL32 is highly conserved among pathogenic Leptospira species. These findings indicate that LipL32 may be important in the pathogenesis, diagnosis, and prevention of leptospirosis.
Acylation↗