Canadian recommendations for laboratory interpretation of multiple or extensive drug resistance in clinical isolates of Enterobacteriaceae, Acinetobacter species and Pseudomonas aeruginosa.

The goal of this document was to provide Canadian laboratories with a framework for consistent reporting and monitoring of multidrug resistant organisms (MDRO) and extensively drug resistant organisms (XDRO) for common gram-negative pathogens. This is the final edition of the interim recommendations, which were modified after one year of broad consultative review. This edition represents a consensus of peer-reviewed information and was co-authored by the Canadian Public Health Laboratory Network and the Canadian Association of Clinical Microbiology and Infectious Diseases. There are two main recommendations. The first recommendation provides standardized definitions for MDRO and XDRO for gram-negative organisms in clinical specimens. These definitions were limited to antibiotics that are commonly tested clinically and, to reduce ambiguity, resistance (rather than non-susceptibility) was used to calculate drug resistance status. The second recommendation identifies the use of standardized laboratory reporting of organisms identified as MDRO or XDRO. Through the broad consultation, which included public health and infection prevention and control colleagues, these definitions are ready to be applied for policy development. Both authoring organizations intend to review these recommendations regularly as antibiotic resistance testing evolves in Canada.

Heterophilic interference in specimens yielding false-reactive results on the Abbott 4th generation ARCHITECT HIV Ag/Ab Combo assay.

BACKGROUND: False-reactivity in HIV-negative specimens has been detected in HIV fourth-generation antigen/antibody or 'combo' assays which are able to detect both anti-HIV-1/HIV-2 antibodies and HIV-1 antigen. OBJECTIVES: We sought to characterize these specimens and determine the effect of heterophilic interference. STUDY DESIGN: Specimens previously testing as false-reactive on the Abbott ARCHITECT HIV Ag/Ab combo assay and re-tested on a different (Siemens ADVIA Centaur HIV Ag/Ab) assay. A subset of these specimens were also pre-treated with heterophilic blocking agents and re-tested on the Abbott assay. RESULTS: Here we report that 95% (252/264) of clinical specimens that were repeatedly reactive on the Abbott ARCHITECT HIV Ag/Ab combo assay (S/Co range, 0.94-678) were negative when re-tested on a different fourth generation HIV combo assay (Siemens ADVIA Centaur HIV Ag/Ab). All 264 samples were subsequently confirmed to be HIV negative. On a small subset (57) of specimens with available volume, pre-treatment with two different reagents (HBT; Heterophilic Blocking Tube, NABT; Non-Specific Blocking Tube) designed to block heterophilic antibody interference either eliminated (HBT) or reduced (NABT) the false reactivity when re-tested on the ARCHITECT HIV Ag/Ab combo assay. CONCLUSIONS: Our results suggest that the Abbott ARCHITECT HIV Ag/Ab combo assay can be prone to heterophilic antibody interference.

Unique combined penA/mtrR/porB mutations and NG-MAST strain types associated with ceftriaxone and cefixime MIC increases in a 'susceptible' Neisseria gonorrhoeae population.

OBJECTIVES: To determine which mutations in penA, mtrR and porB are implicated in increasing minimum MICs of ceftriaxone and cefixime in a susceptible gonococcal population and to ascertain associations with gonococcal strain types (STs). METHODS: One hundred and forty-six Neisseria gonorrhoeae isolates formed two extended-spectrum cephalosporin susceptibility groups: group 1 isolates with cefixime and ceftriaxone MICs of 0.0005-0.016 mg/L; and group 2 isolates with cefixime MICs of 0.03-0.125 mg/L (n = 24) and ceftriaxone MICs of 0.03-0.06 mg/L (n = 23). Mutation patterns in penicillin-binding protein 2 (PBP2; penA), multiple transfer resistance repressor (MtrR; mtrR) and porin B (PorB; porB) were ascertained by DNA sequence and bioinformatic analysis. STs were determined using N. gonorrhoeae multiantigen sequence typing (NG-MAST). RESULTS: Most isolates carried PBP2 mutation pattern IX (D345a, F504L, A510V, A516G and P551L; 50/146, 34.2%), a G45D substitution in MtrR (37.7%) and a wild-type (WT) sequence for PorB (43.2%). Group 2 gonococcal isolates were significantly associated with: penA pattern IX; dual mutations in the promoter (A-) and DNA dimerization domain (H105Y) of MtrR; and G120K;A121D substitutions in PorB. There were 50 combined penA/mtrR/porB mutation patterns, with corresponding patterns I/WT/WT and IX/G45D/G120K;A121D predominating. Gonococci susceptible to ceftriaxone and cefixime were significantly associated with NG-MAST ST 25 (33/36; 92%) and the combined penA/mtrR/porB mutation pattern I/WT/WT. No combined mutation pattern or specific ST was associated with elevated ceftriaxone MICs. NG-MAST ST 3654 was significantly associated with the pattern IX/G45D/G120K;A121D and cefixime group 2 isolates. CONCLUSIONS: Specific single or combined mutation patterns in penA, mtrR and porB and specific STs were associated with differences in susceptibility to ceftriaxone and cefixime.

Comparative evaluation of the Bio-Rad Geenius HIV-1/2 Confirmatory Assay and the Bio-Rad Multispot HIV-1/2 Rapid Test as an alternative differentiation assay for CLSI M53 algorithm-I.

INTRODUCTION: The CLSI-M53-A, Criteria for Laboratory Testing and Diagnosis of Human Immunodeficiency Virus (HIV) Infection; Approved Guideline includes an algorithm in which samples that are reactive on a 4th generation EIA screen proceed to a supplemental assay that is able to confirm and differentiate between antibodies to HIV-1 and HIV-2. The recently CE-marked Bio-Rad Geenius HIV-1/2 Confirmatory Assay was evaluated as an alternative to the FDA-approved Bio-Rad Multispot HIV-1/HIV-2 Rapid Test which has been previously validated for use in this new algorithm. METHODS: This study used reference samples submitted to the Canadian - NLHRS and samples from commercial sources. Data was tabulated in 2×2 tables for statistical analysis; sensitivity, specificity, predictive values, kappa and likelihood ratios. RESULTS: The overall performance of the Geenius and Multispot was very high; sensitivity (100%, 100%), specificity (96.3%, 99.1%), positive (45.3, 181) and negative (0, 0) likelihood ratios respectively, high kappa (0.96) and low bias index (0.0068). The ability to differentiate HIV-1 (99.2%, 100%) and HIV-2 (98.1%, 98.1%) Ab was also very high. CONCLUSION: The Bio-Rad Geenius HIV-1/2 Confirmatory Assay is a suitable alternative to the validated Multispot for use in the second stage of CLSI M53 algorithm-I. The Geenius has additional features including traceability and sample and cassette barcoding that improve the quality management/assurance of HIV testing. It is anticipated that the CLSI M53 guideline and assays such as the Geenius will reduce the number of indeterminate test results previously associated with the HIV-1 WB and improve the ability to differentiate HIV-2 infections.

Classification of Leptospira genomospecies 1, 3, 4 and 5 as Leptospira alstonii sp. nov., Leptospira vanthielii sp. nov., Leptospira terpstrae sp. nov. and Leptospira yanagawae sp. nov., respectively.

The genus Leptospira currently comprises 16 named species. In addition, four unnamed hybridization groups were designated Leptospira genomospecies 1, 3, 4 and 5. These groups represent valid species-level taxa, but were not assigned names in the original description by Brenner et al. [Int J Syst Bacteriol 49, 839-858 (1999)]. To rectify this situation, it is proposed that Leptospira genomospecies 1, genomospecies 3, genomospecies 4 and genomospecies 5 should be classified as Leptospira alstonii sp. nov., Leptospira vanthielii sp. nov., Leptospira terpstrae sp. nov. and Leptospira yanagawae sp. nov., respectively, with strains L. alstonii 79601(T) ( = ATCC BAA-2439(T)), L. vanthielii WaZ Holland(T) ( = ATCC 700522(T)), L. terpstrae LT 11-33(T) ( = ATCC 700639(T)) and L. yanagawae Sao Paulo(T) ( = ATCC 700523(T)) as the type strains. The type strains are also available from the culture collections of the WHO Collaborating Centres in Amsterdam, The Netherlands, and Brisbane, Australia.

A comparison of risk factors associated with community-associated methicillin-resistant and -susceptible Staphylococcus aureus infections in remote communities.

In this case-control study, cases [community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA), n=79] and controls [community-associated methicillin-susceptible S. aureus (CA-MSSA), n=36] were defined as a laboratory-confirmed infection in a patient with no previous hospital-associated factors. Skin and soft tissue were the predominant sites of infection, both for cases (67.1%) and controls (55.6%). Most of the cases (79.7%) and controls (77.8%) were aged <30 years. Investigations did not reveal any significant statistical differences in acquiring a CA-MRSA or CA-MSSA infection. The most common shared risk factors included overcrowding, previous antibiotic usage, existing skin conditions, household exposure to someone with a skin condition, scratches/insect bites, and exposure to healthcare workers. Similar risk factors, identified for both CA-MRSA and CA-MSSA infections, suggest standard hygienic measures and proper treatment guidelines would be beneficial in controlling both CA-MRSA and CA-MSSA in remote communities.

Assessing cost effectiveness of empirical and prophylactic therapy for managing leptospirosis outbreaks.

This study evaluates the utility and cost effectiveness of empirical and prophylactic antibiotic treatment of leptospirosis compared with conventional management. We developed decision trees comparing empirical antibiotic treatment (within 4-7 days of symptom onset) or prophylaxis to conventional antibiotic treatment (initiated 7 days post-onset). Costs were calculated using both US and Barbados pricing. Empirical treatment provided slightly lower probability of survival, while prophylactic treatment resulted in slightly higher survival rates. Antibiotic treatment initiated after 4-7 symptomatic days was ineffective in preventing serious health outcomes, but cost less with the exception of azithromycin (US pricing). Empirical treatment in Barbados cost less than conventional treatment. Prophylaxis reduced rare serious health outcomes and resulted in significant cost savings for the United States and Barbados. Prophylactic therapy for high-risk individuals or prompt diagnosis and early treatment (before 4 days of symptoms) appear to be cost-effective approaches to prevent severe complications of leptospirosis.

Evaluation of three automated nucleic acid amplification systems for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in first-void urine specimens.

A total of 500 first-void urine specimens were tested for the presence of Chlamydia trachomatis and Neisseria gonorrhoeae nucleic acids using ProbeTec ET reagents on a Viper platform (BD Diagnostics, Mississauga, Ontario, Canada), Aptima Combo 2 reagents on a Tigris platform (Gen-Probe, Inc., San Diego, CA), and Abbott RealTime CT/NG reagents on an m2000 platform (Abbott Molecular Diagnostics, Des Plaines, IL). The performance of the three assays for detection of N. gonorrhoeae was comparable, but detection of C. trachomatis by the three assays showed more variation. All three platforms were suitable for the detection of C. trachomatis and N. gonorrhoeae, but additional factors, such as maximum daily specimen throughput, are important in evaluating automated systems for C. trachomatis and N. gonorrhoeae detection in high-volume laboratories.

A quantitative risk assessment of West Nile virus introduction into Barbados / Una evaluación del riesgo cuantitativo de la introducción del virus del Nilo Occidental en Barbados

OBJECTIVE: To present a quantitative risk assessment of West Nile (WNV) virus introduction into Barbados, West Indies. DESIGN AND METHODS: Three possible modes were considered: a) WNV infected mosquitoes via air transport, by city of departure, b) WNV infected mosquitoes via marine transport and c) viraemic migratory, birds. We estimated the number of WNV infected migratory birds as the product of the proportion of migratory birds infected and the number of migratory birds entering Barbados in three taxonomic groups. We further estimated the number of days these birds would be infectious as: [formula: see text]. We then estimated the number (#) of infectious mosquito-days for mosquitoes entering Barbados via air transport as: # infected mosquitoes = (total flights per week/city) x (duration of WNV season) x (number of Culex mosquitoes aboard each flight) x (Culex mosquito WNV infection prevalence) x (vector competence index) x (days infectious). The number of infected mosquitoes entering Barbados via marine transport was estimated using a similar expression as for air transport, except that the number of airplanes and mosquitoes/airplane were substituted with the # of sea containers during a 22-week mosquito season and # of mosquitoes/container. RESULTS: Migratory birds (approximately 69-101 infected birds/year) were associated with the highest introductory risk followed by mode (a) (approximately 2 infected mosquitoes/year) and mode (b) (0. 004 infected mosquitoes/year). CONCLUSIONS: Migratory birds and mosquitoes via air are imminent threats for virus introduction. Impending co-circulation of West Nile virus and four strains of dengue virus may present new challenges for public health.

OBJETIVO: Presentar una valoración del riesgo cuantitativa de la introducción del Virus del Nilo Occidental (VNO) en Barbados, West Indies. MÉTODOS E DISEÑO: Se consideraron tres posibles modos: a) mosquitos infectados con el VNO vía transporte aéreo, por ciudad de salida, b) mosquitos infectados con el VNO vía transporte marítimo, y c) aves migratorias virémicas. Calculamos el número de aves migratorias infectadas con el VNO como el producto de la proporción de aves migratorias infectadas por el número de aves migratorias que entran a Barbados en tres grupos taxonómicos. Luego calculamos el número de días en que estas aves serían infecciosas, de la forma siguiente:[fórmula: ver en el texto].Calculamos entonces el número de días-mosquito infeccioso para los mosquitos que entran en Barbados mediante transporte aéreo, como sigue: # mosquitos infectados = (vuelos totales por semana/ciudad) x (duración de la estación del VNO) x (número de mosquitos Culex a bordo de cada vuelo) x (prevalencia de infección con VNO por mosquito Culex) x (índice de competencia del vector) x (días infecciosos). El número de mosquitos infectados que entraron a Barbados por vía del transporte marítimo fue calculado usando una fórmula similar a la usada en relación con el transporte aéreo, excepto que el número de aeroplanos y mosquitos/ aeroplanos fueron sustituidos con el # de contenedores marítimos durante una temporada de mosquitos de 22 semanas y el # de mosquitos/contenedor RESULTADOS: Las aves migratorias ~ (69-101 aves infectadas/años) estuvieron asociadas con el riesgo de introducción más alto seguido del modo (a) (~2 mosquitos infectados/año), y finalmente el modo (b) (0.004 mosquitos infectados/año). CONCLUSIONES: Las aves migratorias y los mosquitos por vía aérea representan una amenaza inminente de introducción de virus. La co-circulación inminente del Virus del Nilo Occidental y cuatro cepas de virus de dengue pueden presentar nuevos desafíos a la salud pública.

A quantitative risk assessment of West Nile virus introduction into Barbados.

OBJECTIVE: To present a quantitative risk assessment of West Nile (WNV) virus introduction into Barbados, West Indies. DESIGN AND METHODS: Three possible modes were considered: a) WNV infected mosquitoes via air transport, by city of departure, b) WNV infected mosquitoes via marine transport and c) viraemic migratory, birds. We estimated the number of WNV infected migratory birds as the product of the proportion of migratory birds infected and the number of migratory birds entering Barbados in three taxonomic groups. We further estimated the number of days these birds would be infectious as: [formula: see text]. We then estimated the number (#) of infectious mosquito-days for mosquitoes entering Barbados via air transport as: # infected mosquitoes = (total flights per week/city) x (duration of WNV season) x (number of Culex mosquitoes aboard each flight) x (Culex mosquito WNV infection prevalence) x (vector competence index) x (days infectious). The number of infected mosquitoes entering Barbados via marine transport was estimated using a similar expression as for air transport, except that the number of airplanes and mosquitoes/airplane were substituted with the # of sea containers during a 22-week mosquito season and # of mosquitoes/container. RESULTS: Migratory birds (approximately 69-101 infected birds/year) were associated with the highest introductory risk followed by mode (a) (approximately 2 infected mosquitoes/year) and mode (b) (0. 004 infected mosquitoes/year). CONCLUSIONS: Migratory birds and mosquitoes via air are imminent threats for virus introduction. Impending co-circulation of West Nile virus and four strains of dengue virus may present new challenges for public health.

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.

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.

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.

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.

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.

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