Human Bocavirus was detected in 18 (1.5%) of 1,209 respiratory specimens collected in 2003 and 2004 in Canada. The main symptoms of affected patients were cough (78%), fever (67%), and sore throat (44%). Nine patients were hospitalized; of these, 8 (89%) were <5 years of age.

The isolation of human coronavirus NL63 (HCoV-NL63) in The Netherlands raised questions about its contribution to respiratory illness. In this study, a total of 525 respiratory specimens, collected in Canada primarily during the winter months of 2001-2002, were tested for HCoV-NL63; 19 tested positive for HCoV-NL63, demonstrating virus activity during January-March 2002. Patients with HCoV-NL63 were 1 month-100 years old (median age, 37 years). The main clinical presentations were fever (15/19), sore throat (5/19), and cough (9/19), and 4 patients were hospitalized. These results provide evidence for the worldwide distribution of HCoV-NL63.

Human metapneumovirus (hMPV), a newly discovered paramyxovirus, has been associated with acute respiratory tract infections (ARIs) ranging from upper ARIs to severe bronchiolitis and pneumonia. Important questions remain on the contribution of hMPV to ARIs and its impact on public health. During the 2001-2002 season, we conducted a collaborative study with four provincial public health laboratories to study the prevalence of this new virus in the Canadian population. A total of 445 specimens were collected from patients of all age groups with ARIs and were tested for the presence of hMPV by reverse transcription-PCR. Of these, 66 (14.8%) tested positive for hMPV. Positive specimens were found in all age groups and in all four provinces studied. Virus activity peaked in February and March. The age range of the patients with hMPV infection was 2 months to 93 years (median age, 25 years), with similar numbers of females (35%) and males (41%). Thirty-three percent (n ‫؍‬ 22) of hMPV-infected patients were hospitalized; of these, 27% (n ‫؍‬ 6) had rhinitis and pneumonia, 23% (n ‫؍‬ 5) had bronchiolitis, and 9% (n ‫؍‬ 2) had bronchitis. The hospitalization rates were significantly higher among patients <5 years of age (P ‫؍‬ 0.0005) and those >50 years of age (P ‫؍‬ 0.0044) than among those 6 to 50 years of age. Phylogenetic analysis of the F gene showed that two hMPV genetic clusters were cocirculating in the 2001-2002 season, and comparison with earlier studies suggests a temporal evolutionary pattern of hMPV isolates. These results provide further evidence of the importance of hMPV in ARIs, particularly in young children and elderly individuals.

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.Chlamydia trachomatis and Neisseria gonorrhoeae are common bacterial sexually transmitted infections. These infections may remain asymptomatic, particularly in female patients. Untreated chlamydial and gonococcal infections may have longterm sequelae, including pelvic inflammatory disease, ectopic pregnancy, and sterility. The benefits of diagnosis and treatment are well understood. Since the introduction of nucleic acid amplification tests for the detection of C. trachomatis and N. gonorrhoeae in genital tract specimens, this approach has become the standard diagnostic method used in most laboratories. Moreover, urine has also been validated as an adequate specimen for diagnostic testing in many studies (4, 7, 11). We report here on the prospective evaluation of three automated systems for the detection of C. trachomatis and N. gonorrhoeae in urine specimens.A total of 500 first-void urine specimens were tested using ProbeTec ET reagents on the 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 m2000 system is an automated magnetic sample preparation platform combined with homogeneous real-time multiplexed PCR for both C. trachomatis and N. gonorrhoeae (9). All specimens were tested according to the manufacturer's instructions.Each of the assays is designed to detect different targets. The primer sequences are proprietary, but the regions targeted are known. The Abbott RealTime CT/NG assays target the cryptic plasmid of C. trachomatis and the opacity gene of N. gonorrhoeae (9), the ProbeTec ET assays target the cryptic plasmid of C. trachomatis and the pilin-inverting gene of N. gonorrhoeae (11), while the Gen-Probe Aptima Combo 2 detects targets in the 23S rRNA of C. trachomatis and the 16S rRNA of N. gonorrhoeae (3).In order to increase the ability of the study to detect differences in performance between the three assay systems, an additional number of specimens positive for either C. trachomatis or N. gonorrhoeae were included among the 500 specimens. This approach allows th...

A reassortant influenza A(H1N1) virus of swine origin distinct from the pandemic H1N1 2009 strain was isolated from 3 patients, all of whom worked at the same large hog operation in Saskatchewan, Canada. The genomic composition of the isolates has not been previously reported, to our knowledge, and was the product of a genetic reassortment between seasonal H1N1 and triple-reassortant influenza virus that emerged in North American swine during the late 1990s. The neuraminidase and hemagglutinin genes of A/Saskatchewan/5350/2009, A/Saskatchewan/5351/2009, and A/Saskatchewan/5131/2009 were derived from human H1N1 virus and were closely related to those of A/Brisbane/59/2007.

A new rapid direct immunofluorescence assay, the SimulFluor direct fluorescent-antibody (DFA) assay, which can simultaneously detect herpes simplex virus types 1 and 2 (HSV-1 and -2) and varicella-zoster virus (VZV), was evaluated in comparison with our current standard procedures of (i) shell vial direct immunoperoxidase (shell vial IP) staining and cell culture for detection of HSV and (ii) cytospin DFA staining for VZV detection. A total of 517 vesicular, oral, genital, and skin lesion specimens were tested by all three procedures. For HSV detection, the SimulFluor DFA assay had an overall sensitivity, specificity, positive predictive value, and negative predictive value of 80.0, 98.3, 92.3, and 95.1%, respectively, when compared to culture. Shell vial IP staining had a sensitivity, specificity, positive predictive value, and negative predictive value of 87.6, 100, 100, and 96.9%, respectively, when compared with cell culture. The SimulFluor DFA assay, however, offers same-day, 1.5-hours results versus a 1-to 2-day wait for shell vial IP staining results and a 1-to 6-day wait for culture results for HSV. For VZV detection SimulFluor DFA staining detected 27 positive specimens as compared to 31 by our standard cytospin DFA technique-a correlation of 87.1%. A positive SimulFluor reaction for VZV is indicated by yellow-gold fluorescence compared to the bright apple-green fluorescence observed by cytospin DFA staining. There is no difference in turnaround time between the two assays. The SimulFluor DFA assay is a rapid immunofluorescence assay that can detect 80% of the HSV-positive specimens and 87% of the VZV-positive specimens with a 1.5-h turnaround time.Herpes simplex virus (HSV) and varicella-zoster virus (VZV) cause skin lesions in adults and children and may cause severe systemic disease in immunosuppressed hosts and neonates. HSV types 1 and 2 (HSV-1 and -2) can cause vesicular and ulcerative lesions on the genital area as well as oropharyngeal infection. Genital herpes infection is a public health concern, as the infection can be transmitted between sexual partners. Seroprevalence studies of herpes type-specific antibodies have shown an increase of over 30% in the prevalence of HSV-2 infections over the past two decades, with a nationwide incidence of more than 20% of those infected who are 12 years of age and older having detectable antibody to HSV-2 (2). HSV-1 is increasingly recognized as a cause of genital infection, especially in female patients. In the United Kingdom, the annual incidence of HSV-1 genital infection nearly tripled over a 7-period with an incidence of 79% found in one study (8). Most patients with genital herpes infection do not have symptoms and thus are not aware that they can infect their sex partners. Another concern with genital herpes is neonatal herpes. Pregnant women who acquire primary genital herpes shortly before labor are the ones most likely to infect the newborn (1).Herpes zoster virus is a common childhood disease and is also a serious infection in the elderly and imm...

The Syva MicroTrak Chlamydia enzyme immunoassay (EIA; Syva Company, San Jose, Calif.) with cytospin and direct fluorescent-antibody assay (DFA) confirmation was evaluated on 43,630 urogenital specimens over a 1-year period in the Provincial Laboratory in Regina, Saskatchewan, Canada. This was a two-phase study intended to define a testing algorithm for Chlamydia trachomatis that would be both highly accurate and cost-effective in our high-volume (>3,000 tests per month) laboratory. The prevalence of C. trachomatis infection in our population is moderate (8 to 9%). In phase 1, we tested 6,022 male and female urogenital specimens by EIA. All specimens with optical densities above the cutoff value and those within 30% below the cutoff value were retested by DFA. This was 648 specimens (10.8% of the total). A total of 100% (211 of 211) of the specimens with optical densities equal to or greater than 1.00 absorbance unit (AU) above the cutoff value, 98.2% (175 of 178) of the specimens with optical densities of between 0.500 and 0.999 AU above the cutoff value, and 83% (167 of 201) of the specimens with optical densities within 0.499 AU above the cutoff value were confirmed to be positive. A total of 12% (7 of 58) of the specimens with optical densities within 30% below the cutoff value were positive by DFA. In phase 2, we tested 37,608 specimens (32,495 from females; 5,113 from males) by ETA. Only those specimens with optical densities of between 0.499 AU above and 30%o below the cutoff value required confirmation on the basis of data from phase 1 of the study. This was 4.5% of all specimens tested. This decrease in the proportion of specimens requiring confirmation provides a significant cost savings to the laboratory. The testing algorithm gives us a 1-day turnaround time to the final confirmed test results.The MicroTrak ETA performed very well in both phases of the study, with a sensitivity, specificity, positive predictive value, and negative predictive value of 96.1, 99.1, 90.3, and 99.7%, respectively, in phase 2. We suggest that for laboratories that use ETA for Chlamydia testing, a study such as this one will identify an appropriate optical density range for confirmatory testing for samples from that particular population.

There was a high level of agreement between the systems for both CT and GC detection.