Genetic diversity and molecular epidemiology of Norwalk-like viruses.

Specimens (n=287) from 59 gastroenteritis outbreaks collected from February 1997 to March 1999 were analyzed by reverse transcriptase-polymerase chain reaction. The majority of outbreaks (88%) were associated with Norwalk-like viruses. Molecular analyses of strains from 46 outbreaks showed the cocirculation during the 1998-1999 winter of 2 genogroup II clusters, accounting for 57% and 28% of outbreaks, respectively. An important genetic diversity was observed during this 2-year period. Thirteen different genogroup II strains and 3 different genogroup I strains were found. Genogroup I strains, although from the same cluster, were highly divergent (9%-16%). Epidemiologic and molecular data indicate that several introductions did not result in any major shift of prominent strains, whereas 1 apparently established itself. Some point mutations allowed corroboration of epidemiologic links and strongly suggest that, in several instances, sharing staff and/or transfer of patients between health care institutions can create a significant risk for Norwalk-like virus dissemination.

Seroepidemiologic study of three zoonoses (leptospirosis, Q fever, and tularemia) among trappers in Québec, Canada.

This study was undertaken to evaluate the prevalence of antibodies against Francisella tularensis, Coxiella burnetii, and certain serovars of Leptospira interrogans among trappers in Québec, Canada. Muskrat trapping was identified as a risk factor for F. tularensis infection, whereas having a cat at home apparently protected trappers against infection by L. interrogans. High percentages of control sera were positive for antibodies against C. burnetii (15%) and L. interrogans (5%), most frequently serovar bratislava. This is the first report of human infection by serovar bratislava in North America.

Comparison of the fluorescent treponemal antibody absorption (FTA-ABS) test with the FTA-ABS double staining test for detection of antitreponemal immunoglobulin M in the 19S fraction of human serum.

A widely used immunoglobulin M (IgM) detection assay for the diagnosis of neonatal congenital syphilis is the fluorescent treponemal antibody absorption test used with fractionated serum (FTA-ABS 19S IgM test). Reading the results of the FTA-ABS test is more cumbersome than reading those of the FTA-ABS double staining (FTA-ABS-DS) test, a confirmatory test for specific IgG. To verify that the FTA-ABS-DS test used with an anti-human IgM conjugate could detect specific IgM in fractionated serum samples (FTA-ABS-DS 19S IgM test), 164 fractionated (QUIK-SEP IgM Isolation System; ISOLAB, Inc., Akron, Ohio) serum specimens from infected neonates or adults or from IgG-seronegative subjects were tested by both techniques. The sensitivity limits of the two tests were assessed with reactive serum samples diluted to an endpoint titer. Samples nonreactive by the FTA-ABS 19S IgM test (n = 74) were either nonreactive (n = 65), minimally reactive (n = 5), or reactive (n = 4) by the FTA-ABS-DS 19S IgM test. Samples minimally reactive by the FTA-ABS 19S IgM test (n = 32) were minimally reactive (n = 1) or reactive (n = 31) by the double staining test. All samples reactive by the FTA-ABS 19S IgM test (n = 58) were also reactive by the FTA-ABS-DS 19S IgM test. There was a directly proportional linear relationship (r = 0.9794) between titers obtained by both tests. FTA-ABS-DS 19S IgM titers were constantly equal to or higher than FTA-ABS 19S IgM titers. Fluorescence intensity reading repeatability was 91.4% for the FTA-ABS-DS 19S IgM test and 81.7% for the FTA-ABS 19S IgM test (P = 0.015). Because the more easily read FTA-ABS-DS 19S IgM test is at least as sensitive as, if not more sensitive than, the FTA-ABS 19S IgM test, it is a good alternative to the latter test for the detection of specific IgM in human fractionated sera for those using fluorescence microscopes with incident light.

Secondary immune response in a vaccinated population during a large measles epidemic.

The rates of secondary immune response (SIR) and secondary vaccine failure (SVF) during a measles epidemic (10,184 notifications) were evaluated. A patient with SIR was defined as a subject for whom all sera were immunoglobulin G (IgG) positive and IgM negative with a significant increase in complement fixation titer. A patient with SVF was defined as a vaccinated symptomatic subject showing a SIR. Sequential sera from 898 subjects were tested for measles antibody by enzyme-linked immunosorbent assay (IgG and IgM) and by complement fixation. Evidence of recent anti-measles virus specific immune response was found in 496 subjects (55.5%). The vaccination rate was estimated at 74.6% (99% confidence interval [CI], 67.9 to 80.7%). The number of exposed vaccinated subjects was estimated at 370 (74.6% of 496). The SIR rate was 4.03% (20 of 496) (99% CI, 2.1 to 6.9%) among subjects with immune response. These 20 subjects were 2 with measles (Centers for Disease Control's definition), 6 with measles with rash of unknown duration, 8 with presumed measles with either rash or fever, 3 asymptomatic subjects (2 with recent contact with a measles case), and 1 undocumented subject. Since 3 patients with SIR were asymptomatic and 2 others were documented as not vaccinated, there was a maximum of 15 probable occurrences of SVF among the 20 patients with SIR. The SVF rate among exposed vaccinated subjects was estimated at 4.05% (15 of 370) (99% CI, 1.9 to 7.5%). In conclusion, neither prior vaccination nor detectable SIR ensures protective immunity. Measles virus may induce asymptomatic SIR in IgG-seropositive subjects. SVF led to typical or modified measles but did not seem to have played an important role during this epidemic.

Performance and reliability of the Enzygnost measles enzyme-linked immuno-sorbent assay for detection of measles virus-specific immunoglobulin M antibody during a large measles epidemic.

Evaluation of the Enzygnost Measles Enzyme-Linked Immuno-Sorbent Assay kit (Behring) performance to detect specific immunoglobulin M (IgM) was carried out with 3,297 single serum samples and 898 paired serum samples collected during a measles epidemic (10,184 reported cases) in Quebec, Canada. Anti-measles IgM and IgG were detected by using the Enzygnost kit with the appropriate conjugates. Complement-fixing (CF) antibody (Ab) titers were assessed by the laboratory branch complement fixation micromethod. The Centers for Disease Control's clinical measles case definition was used. A modification of the manufacturer's optical density interpretation algorithm was introduced to allow for equivocal results, in addition to positive and negative ones. These three categories differed as to their association with a significant increase in CF Ab titer and the time between the onset of symptoms and phlebotomy. The IgM positivity rate for complement fixation-confirmed measles cases was 96.6% for vaccinated subjects and 100% for nonvaccinated subjects. The daily percentage of IgM seropositivity that was detected for subjects who became IgM positive within 30 days increased gradually from 40 to 90% for sera taken 1 to 7 days after the onset of symptoms, and it plateaued at 100% for sera taken 16 to 30 days after the onset of symptoms. IgM seropositivity was strongly associated with IgG seroconversion, CF Ab titer increase, and clinical measles (P less than 0.0001). Reproducibility was 100% for nonreactive sera and 99.1% for reactive sera. In conclusion, the Enzygnost Measles Enzyme-Linked Immuno-Sorbent Assay kit performed adequately to confirm measles virus infection during this epidemic. A second serum sample should be tested when an early-acute-phase serum sample is IgM negative.

Diagnosing coccidioidomycosis outside an endemic area. A recent case in Montreal, Quebec, Canada.

A 65-year-old female with a long-standing controlled Crohn's disease was admitted in August 1979 to a Montreal hospital for investigation of a non-calcified RUL lung nodule. After an extensive work-up (negative bronchoscopy and mediastinoscopy showing a granulomatous reaction) she underwent thoracotomy; the nodule was enucleated and showed caseous material within a fibrous capsule. Cultures in mycology produced a heavy growth of Coccidioides immitis, a diphasic pathogenic fungus endemic in South Western USA, identified by its morphology in autoclaved cultures, reversion to the spherule phase in mice and a 1:80 immunofluorescent stain with both commercial and patient's serum. The patient, who travelled to Arizona three times in recent years, was soon discharged and remains apparently well. Because of inherent risks from handling cultures of C. immitis, the authors urge that, when coccidioidomycosis is considered clinically and epidemiologically, the pertinent information be transmitted to the laboratory.