Testing human sera for antibodies against avian influenza viruses: horse RBC hemagglutination inhibition vs. microneutralization assays.

BACKGROUND: The hemagglutination inhibition (HI) assay is a frequently used method to screen human sera for antibodies against influenza A viruses. Because HI has relatively poor sensitivity in detecting antibodies against avian influenza A strains, a more complicated microneutralization (MN) assay is often preferred. Recent research suggests that the sensitivity of the HI assay can be improved by switching from the traditionally used turkey, guinea pig, human, or chicken RBCs to horse RBCs. OBJECTIVE: To evaluate the performance of the horse RBC HI when screening for human antibodies against avian influenza types H3, H4, H5, H6, H7, H9, H11, and H12. STUDY DESIGN: We evaluated the reproducibility of horse RBC HI and its agreement with MN results using sera from people exposed or not exposed to wild and domestic birds. RESULTS: The horse RBC HI assay had high reliability (90%-100%) and good agreement with MN assay results (52%-100%). CONCLUSION: The horse RBC HI assay is reliable, less expensive, less complex, and faster than the MN assay. While MN will likely remain the gold standard serologic assay for avian viruses, the horse RBC HI assay may be very useful as a screening assay in large-scale epidemiologic studies.

Infection due to 3 avian influenza subtypes in United States veterinarians.

BACKGROUND: Pandemic influenza virus strains originate in avian species. We examined veterinarians in the United States for evidence of previous avian influenza virus infection. METHODS: We performed a controlled, cross-sectional seroprevalence study among 42 veterinarians and 66 healthy control subjects using serum samples collected from 2002 through 2004. Serum samples were tested using a microneutralization assay against 9 influenza A virus strains. RESULTS: Using multivariable logistic regression modeling, veterinarians exposed to birds demonstrated statistically significant elevated titers against the H5, H6, and H7 avian influenza virus isolates, compared with control subjects. CONCLUSIONS: These data suggest that occupational exposure to avian species may increase veterinarians' risk of avian influenza virus infection. Veterinarians should be considered for priority access to vaccines and antiviral drugs in pandemic planning.

Genotype prevalence and risk factors for severe clinical adenovirus infection, United States 2004-2006.

BACKGROUND: Recently, epidemiological and clinical data have revealed important changes with regard to clinical adenovirus infection, including alterations in antigenic presentation, geographical distribution, and virulence of the virus. METHODS: In an effort to better understand the epidemiology of clinical adenovirus infection in the United States, we adopted a new molecular adenovirus typing technique to study clinical adenovirus isolates collected from 22 medical facilities over a 25-month period during 2004-2006. A hexon gene sequence typing method was used to characterize 2237 clinical adenovirus-positive specimens, comparing their sequences with those of the 51 currently recognized prototype human adenovirus strains. In a blinded comparison, this method performed well and was much faster than the classic serologic typing method. RESULTS: Among civilians, the most prevalent adenovirus types were types 3 (prevalence, 34.6%), 2 (24.3%), 1 (17.7%), and 5 (5.3%). Among military trainees, the most prevalent types were types 4 (prevalence, 92.8%), 3 (2.6%), and 21 (2.4%). CONCLUSIONS: For both populations, we observed a statistically significant increasing trend of adenovirus type 21 detection over time. Among adenovirus isolates recovered from specimens from civilians, 50% were associated with hospitalization, 19.6% with a chronic disease condition, 11% with a bone marrow or solid organ transplantation, 7.4% with intensive care unit stay, and 4.2% with a cancer diagnosis. Multivariable risk factor modeling for adenovirus disease severity found that age <7 years (odds ratio [OR], 3.2; 95% confidence interval [CI], 1.4-7.4), chronic disease (OR, 3.6; 95% CI, 2.6-5.1), recent transplantation (OR, 2.7; 95% CI, 1.3-5.2), and adenovirus type 5 (OR, 2.7; 95% CI, 1.5-4.7) or type 21 infection (OR, 7.6; 95% CI, 2.6-22.3) increased the risk of severe disease.

Are swine workers in the United States at increased risk of infection with zoonotic influenza virus?

BACKGROUND: Pandemic influenza strains originate in nonhuman species. Pigs have an important role in interspecies transmission of the virus. We examined multiple swine-exposed human populations in the nation's number 1 swine-producing state for evidence of previous swine influenza virus infection. METHODS: We performed controlled, cross-sectional seroprevalence studies among 111 farmers, 97 meat processing workers, 65 veterinarians, and 79 control subjects using serum samples collected during the period of 2002-2004. Serum samples were tested using a hemagglutination inhibition assay against the following 6 influenza A virus isolates collected recently from pigs and humans: A/Swine/WI/238/97 (H1N1), A/Swine/WI/R33F/01 (H1N2), A/Swine/Minnesota/593/99 (H3N2), A/New Caledonia/20/99 (H1N1), A/Panama/2007/99 (H3N2), and A/Nanchang/933/95 (H3N2). RESULTS: Using multivariable proportional odds modeling, all 3 exposed study groups demonstrated markedly elevated titers against the H1N1 and H1N2 swine influenza virus isolates, compared with control subjects. Farmers had the strongest indication of exposure to swine H1N1 virus infection (odds ratio [OR], 35.3; 95% confidence interval [CI], 7.7-161.8), followed by veterinarians (OR, 17.8; 95% CI, 3.8-82.7), and meat processing workers (OR, 6.5; 95% CI, 1.4-29.5). Similarly, farmers had the highest odds for exposure to swine H1N2 virus (OR, 13.8; 95% CI, 5.4-35.4), followed by veterinarians (OR, 9.5; 95% CI, 3.6-24.6) and meat processing workers (OR, 2.7; 95% CI, 1.1-6.7). CONCLUSIONS: Occupational exposure to pigs greatly increases workers' risk of swine influenza virus infection. Swine workers should be included in pandemic surveillance and in antiviral and immunization strategies.

Multi-year study of human metapneumovirus infection at a large US Midwestern Medical Referral Center.

BACKGROUND: Because of its recent identification, few multi-year epidemiologic studies of hMPV infection have been reported. OBJECTIVE: We sought to retrospectively describe hMPV infections among patients evaluated by a large US Midwestern referral laboratory. STUDY DESIGN: Clinical specimens were submitted to a large US Midwest referral hospital from 1 October 2001 to 18 May 2004. RT-PCR was used to retrospectively screen the clinical specimens for human metapneumovirus. Demographic and clinical data were retrieved. RESULTS: 34 (2.6%) of 1294 specimens were hMPV positive. Among these, 21 (62%) were culture positive and available for genetic typing. A previously considered rare genotype of hMPV, B1, was the most common single genotype identified, comprising 9 (43%) of the 21 isolates. Multivariate logistic regression modeling identified patients aged 0.4-9 years (OR=8.9; 95% CI=2.0-38.5) and those under intensive care (OR=3.2; 95% CI=1.1-8.7) as more likely to have hMPV infection than their peers. CONCLUSION: In this large referral hospital viral assays more often had evidence of hMPV when they were collected from children receiving intensive care.

Emergent US adenovirus 3 strains associated with an epidemic and serious disease.

BACKGROUND: Adenovirus type 3 (HAdV3) is one of the most prevalent serotypes detected globally. Variants of HAdV3 have been associated with outbreaks of severe disease. OBJECTIVES: To better understand genetic diversity of circulating HAdV3s and examine risk factors for severe disease. STUDY DESIGN: Restriction enzyme analysis for genomic characterization of clinical HAdV3 isolates detected by 15 collaborative US laboratories during the period July 2004 to May 2007. Multivariate modeling was employed for statistical analyses. RESULTS: The most common HAdV3 types of 516 isolates studied were HAdV3a2 (36.9%), HAdV3a50 (27.1%), HAdV3a51 (18.0%), and HAdV3a17 (4.6%). Non-HAdV3a genome types were rare (1.2%). HAdV3a50 and HAdV3a51 are newly described variants which became more prevalent in 2006 and 2007 and have been associated with at least one epidemic. Their uniqueness was determined by specific banding profiles generated by digests with endonucleases BclI, BglII, and HindIII. Multivariable risk factor modeling demonstrated that children under 2 years of age (OR=2.7; 95%CI 1.6-4.6), persons with chronic disease (OR=5.1; 95%CI 2.6-9.8), persons infected with HAdV3a2 (OR=3.0; 95%CI 1.5-6.0), with HAdV3a50 (OR=2.5; 95%CI 1.2-5.2), or with multiple or rare strains (OR=2.8; 95%CI 1.3-6.5) were at increased risk of severe HAdV3 clinical disease. CONCLUSIONS: In the study period considerable genetic diversity was found among US clinical HAdV3 strains. Novel variants emerged and became prevalent. One such emergent strain may be associated with more severe clinical disease.

Evidence for avian influenza A infections among Iowa's agricultural workers.

BACKGROUND: Identifying risk factors for zoonotic influenza transmission may aid public health officials in pandemic influenza planning. OBJECTIVES: We sought to evaluate rural Iowan agriculture workers exposed to poultry for previous evidence of avian influenza virus infection. METHODS: In 2004 we enrolled 803 rural adult Iowans in a 2-year prospective study of zoonotic influenza transmission. Their enrollment data and sera were compared with those from 66 adult controls enrolled at the University of Iowa in 2006 by proportional odds modeling. RESULTS: The 803 participants were 58.8% male with a mean age of 55.6 yrs. Forty-eight percent reported previous poultry exposure. Sera were studied by microneutralization techniques for antibodies against avian H4, H5, H6, H7, and H9 viruses. Touching live birds was associated (OR = 1.2; 95% CI 1.02-1.8) with increased antibody titer against H5 virus. Similarly, participants who reported hunting wild birds had increased antibody titers against H7 virus (OR = 2.8; 95%CI = 1.2-6.5) and subjects who reported recent work with poultry had increased antibody titers against H6 (OR = 3.4; 95% CI 1.4-8.5) and H7 viruses (OR = 2.5, 95% CI = 1.1-5.7). There was no evidence of elevated antibody against avian H4 or H9 viruses. CONCLUSIONS: These data suggest that hunting and exposure to poultry may be important risk factors for avian influenza virus infection among rural US populations. Agriculture workers should be included in influenza pandemic plans.

Swine workers and swine influenza virus infections.

In 2004, 803 rural Iowans from the Agricultural Health Study were enrolled in a 2-year prospective study of zoonotic influenza transmission. Demographic and occupational exposure data from enrollment, 12-month, and 24-month follow-up encounters were examined for association with evidence of previous and incident influenza virus infections. When proportional odds modeling with multivariable adjustment was used, upon enrollment, swine-exposed participants (odds ratio [OR] 54.9, 95% confidence interval [CI] 13.0-232.6) and their nonswine-exposed spouses (OR 28.2, 95% CI 6.1-130.1) were found to have an increased odds of elevated antibody level to swine influenza (H1N1) virus compared with 79 nonexposed University of Iowa personnel. Further evidence of occupational swine influenza virus infections was observed through self-reported influenza-like illness data, comparisons of enrollment and follow-up serum samples, and the isolation of a reassortant swine influenza (H1N1) virus from an ill swine farmer. Study data suggest that swine workers and their nonswine-exposed spouses are at increased risk of zoonotic influenza virus infections.

Avian influenza and poultry workers, Peru, 2006.

BACKGROUND: Currently numerous countries in Asia, Africa and Europe are encountering highly pathogenic avian influenza (AI) infections in poultry and humans. In the Americas, home of the world's largest poultry exporters, contingency plans are being developed and evaluated in preparation for the arrival of these viral strains. OBJECTIVES: With this cross-sectional study, to our knowledge the first in its kind in Central or South America, we sought to learn if Peruvian poultry workers had evidence of previous AI infection and if so, to determine risk factors for infection. METHODS: We performed a cross-sectional seroprevalence study among 149 workers on a Peruvian poultry farm (133 exposed to poultry and 17 non-exposed controls), serum samples were tested for human influenza virus exposure using a hemagglutination inhibition (HI) assay. Microneutralization assays were performed on all serum samples to detect antibodies against prototypic avian influenza (AI) strains H4 through H12. RESULTS: Using multivariate proportional odds modeling we found that the prevalence of elevated titers against AI viruses was low in both groups, exposed and non-exposed controls. CONCLUSIONS: No evidence of previous avian influenza infection among Peruvian poultry workers was found in this first cross-sectional study performed in South America. This first occupational study of AI in Latin America was encouraging, but it likely reflects the sector of poultry production with higher biosecurity.

Preventing zoonotic influenza virus infection.

We evaluated 49 swine industry workers and 79 nonexposed controls for antibodies to swine influenza viruses. Multivariate modeling showed that workers who seldom used gloves (odds ratio [OR] 30.3) or who smoked (OR 18.7) most frequently had evidence of previous H1N1 swine virus. These findings may be valuable in planning for pandemic influenza.

Human metapneumovirus, Peru.

We retrospectively studied 420 pharyngeal swab specimens collected from Peruvian and Argentinean patients with influenzalike illness in 2002 and 2003 for evidence of human metapneumovirus (HMPV). Twelve specimens (2.3%) were positive by multiple assays. Six specimens yielded HMPV isolates. Four of the 6 isolates were of the uncommon B1 genotype.

Emergent strain of human adenovirus endemic in Iowa.

We evaluated 76 adenovirus type 7 (Ad7) isolates collected in Iowa from 1992 to 2002 and found that genome type Ad7d2 became increasingly prevalent. By 2002, it had supplanted all other Ad7 genome types. The association of Ad7d2 with severe illness and death calls for heightened public health concern.