Gregory W. Hammond

Antimicrobial Susceptibility of Haemophilus ducreyi

The susceptibility of 19 isolates of Haemophilus ducreyi from a recent chancroid outbreak and four reference strains was determined in vitro to 13 antimicrobial agents. The rabbit intradermal test for virulence was positive for all of the local isolates, but not for the reference strains. The “nonvirulent” reference strains were inhibited by lower minimum inhibitory concentrations (MICs) of most agents tested. For the virulent isolates, the range of MICs (in micrograms per milliliter) of the following were: of vancomycin, 8 to 128; of polymyxin, 32 to 128; of cloxacillin, 32 to 64; of tetracycline, 0.5 to 32; of cephalothin, 4 to 8; of doxycycline, 0.25 to 8; and of kanamycin, 1 to 8. Three strains were resistant to penicillin and ampicillin (MIC ≥ 128 μg/ml), and these three strains produced β-lactamase. The remainder were susceptible to 4 μg/ml. All strains were susceptible to rifampin (MIC ≤ 1 μg/ml), chloramphenicol (MIC ≤ 4 μg/ml), sulfisoxazole (MIC ≤ 8 μg/ml), and nalidixic acid (MIC ≤ 8 μg/ml). These susceptibilities of H. ducreyi indicate several antimicrobial agents that may be effective for chancroid treatment and support the use of vancomycin in a selective medium for the culture of chancroid genital ulcers. Images

Effectiveness of the pandemic H1N1 influenza vaccines against laboratory-confirmed H1N1 infections: population-based case-control study.

BACKGROUND Excellent immune responses following 1 or 2 doses of the monovalent inactivated pandemic H1N1 vaccines have been documented, but the effectiveness of these vaccines against laboratory-confirmed H1N1 infections in the general population is not clear. We evaluated the effectiveness of the pandemic H1N1 and seasonal trivalent influenza vaccines (TIV) used during the 2009 mass vaccination campaign in Manitoba (Canada) in preventing laboratory-confirmed H1N1 infections. METHODS A population-based case-control study using data from Cadham Provincial Laboratory (CPL) and the Manitoba Immunization Monitoring System (MIMS). All Manitoba residents ≥6 months of age who had a respiratory specimen tested at CPL for H1N1 were included in the study. Cases were individuals who tested positive for pandemic H1N1 influenza A by reverse transcriptase-PCR (N=1435). Controls were individuals who tested negative for both influenza A and B (N=2309). Information on receipt of TIV or H1N1 vaccine was obtained by record linkage with MIMS, the population-based province-wide immunization registry. RESULTS Overall, the adjuvanted H1N1 vaccine was 86% (95%CI 75-93%) effective in preventing laboratory-confirmed H1N1 infections when vaccination occurred ≥14 days before testing. Effectiveness seemed lower among older (≥50 years) individuals [51% (-51 to 84%)] and among those with immunocompromising conditions [67% (-13 to 90%)]. There was also evidence that the H1N1 vaccine might be less effective among those who had received the 2009/10 TIV. DISCUSSION The adjuvanted H1N1 vaccine used during Manitobas H1N1 mass vaccination campaign was highly effective against laboratory-confirmed pandemic H1N1 infection, especially among children and younger adults.

Bacterial antigen detection in cerebrospinal fluid of patients with meningitis

This study compared the sensitivity and specificity of four test systems in detecting Haemophilus influenzae type b, Neisseria meningitidis, Streptococcus pneumoniae, and gram-negative organisms in cerebrospinal fluid (CSF), versus culture. The tests used on CSF from 155 patients with meningitis were the Phadebact coagglutination (CoA) test, the Directigen latex agglutination (LA) test, counterimmunoelectrophoresis (CIE), and the Limulus amebocyte lysate (LAL) test. The sensitivity for patients with bacterial meningitis was 78% (18/23) for LA, 78% (25/32) for CoA, and 67% (18/27) for CIE for detection of H. influenzae type b; 71% (10/14) for CoA, 100% (6/6) for LA, and 50% (6/13) for CIE in detecting S. pneumoniae; and 33% (1/3) for LA and 50% (2/4) for CIE in detecting N. meningitidis. LAL had a sensitivity of 77% (37/48) in detecting CSF gram-negative endotoxin. The specificities of those with bacterial meningitis for H. influenzae, S. pneumoniae, and N. meningitidis tested by LA were, respectively, 100% (35/35), 96% (50/52), and 100% (54/54); for H. influenzae and S. pneumoniae using CoA 97% (62/64) and 96% (80/83); for H. influenzae, S. pneumoniae, and N. meningitidis using CIE 67% (18/27), 50% (6/12), and 50% (2/4). The specificity of LAL was 86% (38/44). The detection of bacterial antigen from CSF in patients with meningitis by commercial agglutination tests is more sensitive than CIE and is highly specific.

COMPARISON OF IMMUNOGENICITY OF TWO YEAST-DERIVED RECOMBINANT HEPATITIS B VACCINES

A comparison was conducted of the immunogenicity of two yeast recombinant vaccines with different doses [10 micrograms Recombivax-HB (Merck Sharpe & Dohme Ltd) vs. 20 micrograms Engerix-B (Smith Kline Biologicals)]. This was conducted as a randomized, blinded study in healthy preclinical medical students, negative for hepatitis B markers. The geometric mean titres (GMT) showed a wide individual variability for both vaccines, and approximately a two- to threefold greater GMT of anti-HBs in recipients of the 20 micrograms vaccine. However, the 95% confidence interval showed an overlap of the means of the GMT for both vaccine groups, and in this study there was no significant difference in immunogenicity of these two vaccines. At 6-7 months after completion of immunization, antibody levels fell to one-third of the levels of one month post-immunization. A case report of an allergic urticarial reaction to one of the recombinant vaccines is presented.

No association between 2008-09 influenza vaccine and influenza A(H1N1)pdm09 virus infection, Manitoba, Canada, 2009.

Receipt of seasonal inactivated trivalent vaccine neither increased nor decreased the risk for pandemic influenza virus infection.

Comparison of cell culture and three enzyme-linked immunosorbent assays for the rapid diagnosis of respiratory syncytial virus from nasopharyngeal aspirate and tracheal secretion specimens

A total of 211 specimens, including 144 nasopharyngeal aspirates and 67 tracheal secretions, were evaluated for the rapid detection of Respiratory Syncytial Virus (RSV) antigen by three commercial enzyme-linked immunosorbent assays (ELISA; Kallestad Diagnostic, Austin, TX; Ortho Diagnostics, Raritan, NJ, and Abbott Laboratories, North Chicago, IL) and by isolation of RSV in cell culture. Of the 61 RSV culture positive specimens, Kallestad ELISA, Ortho ELISA, and Abbott ELISA detected RSV antigen in 80%, 95%, and 92% of the specimens, respectively. An additional 28 specimens were found to be positive only by one or more RSV ELISA tests. A blocking assay confirmed the specificity of ELISA in 71% (20/28) of the RSV ELISA positive and the culture-negative specimens, and 29% were found to be false positive. Through the use of cell culture, with the resolution of ELISA positive and culture negative specimens by blocking assay, 81 specimens (61 + 20) were considered to be true positive. The sensitivity, specificity, and diagnostic accuracy were, for cell culture, 75%, 100%, and 91%; for Kallestad ELISA, 79%, 98%, and 91%; for Ortho ELISA, 95%, 99%, and 98%; and for Abbott ELISA, 93%, 96%, and 95% respectively. In our study, commercial ELISA tests have been shown to be rapid, reliable tests for the detection of RSV. Ortho ELISA and Abbott ELISA showed better sensitivity than the Kallestad ELISA for RSV detection directly in the clinical specimens.

Two cases of Hemophilus endocarditis of prolapsed mitral valves—Hemophilus paraphrophilus or parainfluenzae?

Abstract Documented here are the clinical and laboratory features in two recent cases of subacute bacterial endocarditis caused by organisms of the Hemophilus species. Classifications of the suspected pathogen involved are reviewed with an emphasis on the distinction between Hemophilus paraphrophilus and the closely related organism, Hemophilus parainfluenzae. The two isolates described herein have been tentatively identified as H. paraphrophilus based on Zimmermans original description—also used for other recent reports of infection due to this organism.

Comparison of Detection Methods for Adenovirus from Enteric Clinical Specimens

Abstract Fecal samples submitted for virus examination over July 1990 to June 1991 from children <3 years of age were examined by electron microscopy (EM), virus culture (VC), and enzyme immunoassay [EIA, group-reactive and adenovirus (Ad) 40/41 specific; Cambridge BioScience] to compare the detection rate of adenovirus from pediatric fecal specimens. Ad isolates of serotypes 1–7 grown in HEp-2 or primary rhesus monkey kidney cells were identified by neutralization. Graham 293 cell cultures were used only when specimens were found to be positive for Ad by EM, type-specific Ad40/41 EIA, and for isolates not identified by neutralization. Ads grown in 293 cells were identified by DNA restriction endonuclease analysis. Of the 1187 specimens examined, 105 (9%) were found to be positive for Ad. VC detected 93, while 12 additional positives were detected by EM or EIA. The relative sensitivity of VC, EIA, and EM for the 105 specimens was 89% (93), 45% (47), and 35% (37), respectively. Among the 105 positive specimens, enteric Ad, nonenteric Ad, and untypeable Ad were 28% (29), 65% (68), and 7% (8), respectively. Of 37 EM positives, 62% (23) were enteric Ad; 27% (10) were nonenteric including serotypes 2, 3, 4, 5, 12, and 31, with 4, 1, 1, 2, 1, and 1 isolates of each type positive, respectively; and 11% (4) were detectable only by EM. Five isolates were identified as variant of Ad 2(3), Ad 3(1) and Ad 31(1). Over a 1-year period, a single Ad41 variant strain was the most frequently detected enteric Ad in Winnipeg, Manitoba, Canada. For maximum detection rates of Ad viruses in pediatric fecal specimens, a combination of EM, VC, and EIA is required, but group-reactive EIA, or EM followed by Ad40/41-specific EIA of initial positives, are the most direct and efficient methods for enteric Ad detection.

Restriction analysis of the prototype strain of enteric adenovirus type 41 using exonuclease III

Enteric adenoviruses 40 and 41 (Ad40 and Ad41) are a prominent cause of gastroenteritis in young children. Diagnosis of these enteric types by conventional means is complicated by their fastidious growth characteristics. Enteric adenovirus growth was enhanced by cocultivation. Typing of enteric isolates currently entails analysis of the extracted viral DNA with restriction enzymes. Restriction endonuclease fragments of the Ad41 strain Tak genome were ordered by (i) double digestion, (ii) release of restriction fragments from plasmids containing 84% of the Ad41 genome in EcoRI fragments A, B and C, (iii) hybridization of Southern blotted Ad41 fragments with EcoRI fragment containing plasmids and various segments of the Ad2 genome, (iv) sequential reduction of the genome beginning with terminal restriction fragments with exonuclease III and S1 nuclease. The termini of adenovirus genomes are difficult to clone and the use of exonuclease III is a useful alternative to conventional restriction mapping. DNA restriction patterns, fragment sizes and restriction maps of the Ad4 1 strain Tak with enzymes BamHI, BglII, ClaI, EcoRI, HindIII, PstI, SalI, SmaI and XhoI are presented. Prototype strain restriction maps should enable better understanding of adenovirus type 41 and its epidemiology.

Did the H1N1 Vaccine Reduce the Risk of Admission with Influenza and Pneumonia during the Pandemic

Background The extent to which A(H1N1)pdm09 influenza vaccines prevented hospital admissions with pneumonia and influenza (P&I) during the 2009 pandemic remains poorly understood. We evaluated the effectiveness of the A(H1N1)pdm09 and seasonal influenza vaccines (TIV) used during the 2009 mass vaccination campaign in Manitoba (Canada) in preventing P&I hospitalization. Methods A population-based record-linkage nested case-control study. Cases (N = 1,812) were persons hospitalized with influenza (ICD-10:J09-J11) or pneumonia (ICD-10:J12-J18) during the study period. Age-, gender- and area of residence-matched controls (N = 7,915) were randomly sampled from Manitoba’s Population Registry. Information on receipt of A(H1N1)pdm09 vaccine and TIV was obtained from the Manitoba Immunization Monitoring System, a province-wide vaccine registry. Results Overall, the adjuvanted A(H1N1)pdm09 vaccine was 27% (95%CI 13–39%) effective against P&I hospitalization ≥ 14 days following administration. Effectiveness seemed lower among older (≥ 65 years) adults (10%; −16–30%), particularly when compared to under-5 children (58%; 30–75%). The number-needed-to-vaccinate to prevent 1 P&I admission was lowest among <4 year-olds (928) and ≥65 years (1,721). VE against hospitalization with laboratory-confirmed A(H1N1)pdm09 was 70% (39–85%) overall and (91%; 62–98%) ≥ 14 days following vaccination. Discussion Our data suggest that the adjuvanted A(H1N1)pdm09 vaccine was effective in preventing about 55–60% of P&I hospitalizations among children and younger adults who were at much higher risk of infection. Unfortunately, the vaccine was less effective among 65 or older adults. Despite that the vaccine still had a significant population-based impact especially among the very young (<5) and the older (≥ 65 years).