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Biomedical subjects

J E Herrmann

Publications and source records attributed to J E Herrmann.

At least 55 records · Page 3Linked to original sources

Incidence of enteric adenoviruses among children in Thailand and the significance of these viruses in gastroenteritis.

In countries with temperate climates, enteric adenoviruses have been shown to be a substantial cause of pediatric gastroenteritis. To determine the incidence of adenovirus infection in a tropical climate, stools were collected from children under age 7 during a 1-year period at an outpatient clinic in Bangkok, Thailand. Stools from 1,114 children with gastroenteritis and from 947 children without gastroenteritis were tested. Each stool was tested for adenovirus group antigen and for specific enteric adenovirus types (Ad40 and Ad41) by monoclonal antibody enzyme immunoassays. We found that 4.4% (49 of 1,114) of children with gastroenteritis and 1.8% (17 of 947) of children without gastroenteritis were positive for adenovirus group antigen. In tests for specific enteric adenovirus types, 2.0% (22 of 1,114) of the tests were positive in children with gastroenteritis and 0.6% (6 of 947) were positive in children without gastroenteritis. There was a significant correlation (P less than 0.02) of gastroenteritis with nonenteric adenovirus types (27 of 1,114) as well as with specific enteric adenovirus types (P less than 0.01). By comparison, 19.7% of children with gastroenteritis and 0.7% of those without gastroenteritis were positive for rotavirus infection. In the adenovirus-infected children with gastroenteritis, there were coinfections with rotavirus only in those with nonenteric adenovirus infection (7 of 27 children). There were no significant differences in the association of bacterial or parasitic infections with either enteric or nonenteric adenovirus infections in either group of children studied. These data demonstrate that Ad40 and Ad41 are causes of gastroenteritis in this population, but among the spectrum of diarrheal etiologies, they may be proportionately less important than they are in countries with temperate climates.

Adenoviridae Infections↗

Immunobiology of Norwalk virus.

Clinical immunity to Norwalk virus in inoculated human volunteers appears to be unusual for gastroenteritis viruses, as certain individuals are repeatedly ill on long-term virus rechallenge and others remain persistently well. In these volunteers there is a paradoxical inverse correlation between the prechallenge serum (and jejunal fluid) Norwalk antibody level (measured by radioimmunoassay) and resistance to illness, suggesting that non-immunological factors, perhaps genetic, may be important in determining resistance. Most reported naturally occurring Norwalk disease outbreaks in developed nations also show that humoral antibody fails to correlate with immunity to infection. The unusual pattern of clinical immunity to Norwalk virus indicates a need for caution in the development of vaccines against this agent as well as a need for additional information on its immunobiological characteristics. The virus is known to contain a single protein, like the caliciviruses. Recently we have found evidence for at least a one-way serological cross-relatedness between Norwalk virus and human calicivirus. Twelve of 20 paired sera from ill patients in outbreaks due to calicivirus strain UK4 seroconverted to Norwalk virus by radioimmunoassay and two of eight paired sera from UK2 outbreaks showed seroconversion. Future studies of outbreaks caused by various calicivirus strains should be designed to correlate acute-phase serum antibody titres to Norwalk virus with clinical susceptibility and immunity to infection.

Antibodies, Viral↗

Preparation and characterization of monoclonal antibodies to enteric adenovirus types 40 and 41.

We have prepared monoclonal antibodies to each of the enteric adenoviruses types 40 and 41. Three different hybridoma cell lines were selected which produced antibody found to react by radioimmunoprecipitation with adenovirus (Ad) hexon antigens. One was specific for Ad 40, another for Ad 41, and a third one reacted with both types. When tested in an enzyme immunoassay against all 41 known human Ad types, the type-specific monoclonal antibody against Ad 40 reacted homotypically, as did the monoclonal antibody against Ad 41. In addition, these monoclonal antibodies neutralized the homologous enteric Ad type. The monoclonal antibody which reacted with both enteric Ad types also showed lower levels of reactivity with the group C adenoviruses types 2, 5, and 6. The monoclonal antibodies produced will provide a definitive means for rapid identification of specific Ad types, and will be useful in defining the relationship of enteric adenoviruses to other types.

Adenoviruses, Human↗

Antigen detection with monoclonal antibodies for the diagnosis of adenovirus gastroenteritis.

A monoclonal antibody-based enzyme immunoassay (EIA) was developed for direct detection of enteric adenoviruses in stool specimens from individuals with gastroenteritis. Tests specific for each of the enteric adenoviruses, adenovirus type 40 (Ad 40) and type 41 (Ad 41), were designed. The sensitivity of the assay was determined by comparing the results of the EIA with isolation of virus in Graham 293 cells from stools that contained particles having adenovirus morphology. The standard for specificity was analysis of adenovirus genome profiles after digestion with SmaI endonuclease. The sensitivity was 95.8% (23 of 24) for Ad 40 and 97.1% (34 of 35) for Ad 41. The specificity was 95.7% (45 of 47) and 97.2% (35 of 36), respectively. The two type-specific monoclonal antibodies could be mixed in an EIA for identification of enteric adenoviruses in stools without loss of reactivity in either type. The EIA permits rapid diagnosis and type-specific identification of enteric adenoviruses in gastroenteritis.

Adenoviridae Infections↗

Antigenic relationships between human caliciviruses and Norwalk virus.

We investigated antigenic relationships between human calicivirus (HCV) strains and Norwalk virus by using immune electron microscopy (IEM) and radioimmunoassay (RIA). Three serologically distinct HCV strains, UK1, UK2, and Japan, were demonstrated by IEM, as was evidence for two additional strains, UK3 and UK4. Although HCV strains and Norwalk virus were distinct by IEM, 12 of 20 patients with gastroenteritis due to HCV UK4 and two of eight with gastroenteritis due to UK2 showed seroconversions to Norwalk virus by RIA. These naturally occurring antibody responses in humans, as detected by RIA, support the concept that Norwalk virus belongs to the family Caliciviridae. An RIA for HCV Japan antigen also detected HCV UK1, UK2, and UK4 and thus appears to identify a group-specific antigen for these viruses. An RIA for antibody to HCV Japan failed to identify seroconversions in 45 of 47 patients with gastroenteritis due to HCV UK. These results may reflect different reactivities of various immunologic tests for the identification of infections due to small gastroenteritis viruses.

Animals↗

Foodborne Snow Mountain agent gastroenteritis in a school cafeteria.

In 1984, an outbreak of gastroenteritis occurred at a school with 1,860 students in Brooklyn, NY. In a single-stage cluster sample of 375 students, 129 (34%) had illnesses that met our case definition of vomiting or diarrhea. The mean incubation period was 26 hours, and the mean illness duration was 24 hours. All case students had eaten in the cafeteria on at least one day between Nov 13 and 16, compared with 174/214 (81%) noncase students (P = 10(-8), Fisher exact test). Foods implicated were french fries (relative risk 1.7, 95% confidence limits 1.4, 2.0) and hamburgers (relative risk 1.6, 95%, confidence limits 1.2, 2.1). Two cafeteria employees had served those foods while affected by diarrhea. By a recently developed blocking enzyme-linked immunosorbent assay, six of 11 (55%) case students showed fourfold antibody increases between acute- and convalescent-phase serum samples for Snow Mountain agent, a Norwalk-like virus, compared with one of ten (10%) noncase students (P = .04, Fisher exact test). We strongly suspect, but cannot document conclusively, that the Snow Mountain agent was spread to students on a vector of hot foods contaminated by ill food handlers. Implicated foods conferred low relative risks and could only have accounted for 74% of cases of illness. The strong association between cafeteria exposure and illness, therefore, suggests that additional modes of spread occurred.

Antibodies, Viral↗

Widespread outbreaks of clam- and oyster-associated gastroenteritis. Role of Norwalk virus.

Consumption of raw shellfish has long been known to be associated with individual cases and sporadic outbreaks of enteric illness. However, during 1982, outbreaks of gastroenteritis associated with eating raw shellfish reached epidemic proportions in New York State. Between May 1 and December 31, there were 103 well-documented outbreaks in which 1017 persons became ill: 813 cases were related to eating clams, and 204 to eating oysters. The most common symptoms were diarrhea, nausea, abdominal cramps, and vomiting. Incubation periods were generally 24 to 48 hours long, and the duration of illness was 24 to 48 hours. Bacteriologic analyses of stool and shellfish specimens did not reveal a causative agent. Norwalk virus was implicated as the predominant etiologic agent by clinical features of the illness and by seroconversion and the formation of IgM antibody to Norwalk virus in paired serum samples from persons in five (71 percent) of seven outbreaks in which testing was done. In addition, Norwalk virus was identified by radioimmunoassay in clam and oyster specimens from two of the outbreaks. Determining the source of the shellfish was not always possible, but northeastern coastal waters were implicated. The magnitude, persistence, and widespread nature of these outbreaks raise further questions about the safety of consuming raw shellfish.

Antibodies, Viral↗

Detection of rotavirus with a new polyclonal antibody enzyme immunoassay (Rotazyme II) and a commercial latex agglutination test (Rotalex): comparison with a monoclonal antibody enzyme immunoassay.

A total of 176 human fecal specimens were examined for the presence of rotavirus by four different assays: a monoclonal antibody enzyme immunoassay; the original polyclonal antibody enzyme immunoassay marketed by Abbott Laboratories, North Chicago, Ill. (Rotazyme I); a modification of this assay which is now commercially available (Rotazyme II); and a latex agglutination test (Rotalex) recently introduced by Medical Technology Corp., Somerset, N.J. In addition, selected specimens were examined for the presence of rotavirus by electron microscopy, immune electron microscopy, and RNA gel electrophoresis. A total of 40 specimens were positive in the monoclonal antibody enzyme immunoassay, and 136 were negative. Using the results obtained with this procedure as the reference standard, we found the sensitivities of the Rotazyme I, Rotazyme II, and Rotalex tests to be 97.4, 100, and 81.6%, respectively. The specificities of these three procedures were 88.8, 83.9, and 100%, respectively.

Antibodies, Monoclonal↗

Evaluation of chlamydiazyme for the detection of genital infections caused by Chlamydia trachomatis.

Chlamydiazyme is a 4-h enzyme-linked immunoassay that detects an antigen of Chlamydia trachomatis directly in clinical specimens. This immunoassay was compared with cell culture for the diagnosis of chlamydial infections of the genital tract. The assay was evaluated at five clinics with a total of 1,277 cervical specimens of which 239 were culture positive. At three of these clinics where urethral samples were taken from males, 99 of 363 samples were culture positive. The sensitivity of the assay averaged 89.5% for detecting cervical infections and 78.8% for detecting male urethral infections. Specificity was 97.0% when samples from either males or females were tested. Some patients who were culture negative were infected with chlamydiae according to both Chlamydiazyme and a monoclonal antibody test that detected a chlamydial antigen distinct from the antigen detected by Chlamydiazyme. If the 15 females and 2 males who were positive by both immunoassays but culture negative were considered positive for chlamydial infection, the specificity of the assay was 98.4% in females and 97.7% in males. Chlamydiazyme is a simple and relatively rapid immunoassay that has sufficient sensitivity and specificity to supplant culture in the detection of genital chlamydial infections.

Antigens, Bacterial↗

Detection of Norwalk virus in stools by enzyme immunoassay.

The development of a solid-phase microtiter enzyme immunoassay (EIA) for detection of Norwalk virus antigen in stool samples is described. The EIA was compared with a previously developed radioimmunoassay (RIA) for detection of Norwalk virus antigen in stools obtained from 30 volunteers who received Norwalk virus. The EIA detected viral antigen in stools from 17 of the volunteers and the RIA detected viral antigen in 15. Seroconversion was a more sensitive indicator of infection in some patients. However, two samples from volunteers who were clinically ill but did not show seroconversion to Norwalk virus were positive for Norwalk virus antigen by both immunoassays. This indicates that antigen detection may be important for use in epidemiological studies. Neither of the immunoassays gave positive reactions for stools known to contain enteric adenovirus, rotavirus, or Hawaii virus, or in stools from patients with acute diarrhea of unknown cause. The stability of the EIA reagents and ease of use should provide a means for more extensive testing for Norwalk virus in outbreaks of gastroenteritis.

Antigens, Viral↗

Antigen detection for the diagnosis of gonorrhea.

We compared the diagnostic value of an enzyme immunoassay method for detection of gonococcal antigen in genital secretions with culture results and direct Gram stain for Neisseria gonorrhoeae in 1,171 men and 723 women attending a sexually transmitted disease clinic. When compared with culture results in men, the immunoassay provided a sensitivity of 94% and a specificity of 98% and was essentially equivalent to the urethral Gram stain. The predictive value of a positive immunoassay was 97% in men with a urethral discharge in whom the prevalence of gonorrhea was 36%, and 30% in men without urethral discharge, who had a 2% prevalence of gonorrhea (P less than 0.001). The sensitivity of the immunoassay was 95% in men with and 67% in men without urethral discharge (P less than 0.01). In women, the immunoassay resulted in a sensitivity of 78% and a specificity of 98% compared with cervical culture and had a significantly better sensitivity than the cervical Gram stain (78 versus 48%, P less than 0.001). Analysis of patients with discrepant culture and immunoassay results suggested that most culture-negative, immunoassay-positive patients probably did not have gonorrhea. After treatment, all but 1 of 59 originally culture- and immunoassay-positive patients became negative in both tests by 3 days. Results of the immunoassay were not affected by transport or by refrigeration for up to 30 days.

Antigens, Bacterial↗

Detection of Chlamydia trachomatis in genital specimens by the Chlamydiazyme test.

Cotton swabs were used to collect two specimens each from 416 patients (206 males, 210 females) attending a sexually transmitted disease clinic. The first swab was transported in Specimen Storage Reagent and extracted in Specimen Dilution Buffer for enzyme immunoassay by Chlamydiazyme (Abbott Laboratories); the second swab was extracted into 2SP and inoculated into McCoy cell cultures. In the first phase of the study (215 patients: 111 males, 114 females) enzyme immunoassay results were positive (optical density greater than or equal to 0.1) in 30 of 35 instances in which Chlamydia trachomatis was isolated (sensitivity, 86%). Of 18 false-positive enzyme immunoassay results, 15 (83%) were cervical swabs (specificity, 90%). In a phase II study, using a modified Chlamydiazyme kit, 201 patients were tested (95 males, 106 females). Of 41 chlamydial isolates, 8 were not detected by the Chlamydiazyme test (sensitivity, 81%). Only three positive Chlamydiazyme test results could not be confirmed by culture (specificity, 98%). Overall, Chlamydiazyme assay provided a rapid (4 h), sensitive, and specific assay for the detection of chlamydial antigens.

Antigens, Bacterial↗

Detection and identification of influenza virus antigens by nylon-coupled enzyme-linked immunoassay.

A direct solid-phase enzyme-linked immunoassay for rapid detection and typing of influenza virus was developed utilizing antibodies immobilized by covalent linkage to nylon beads. Covalent linkage of antibody to nylon was accomplished by treatment of partially hydrolyzed nylon with glutaraldehyde. For comparison to conventional enzyme-linked immunosorbent assays (ELISA), IgG fractions were adsorbed to polystyrene beads. Influenza type-specific immunoglobulins coupled to nylon beads were used in an enzyme-linked immunoassay to identify influenza A/USSR/77(H1N1), and A/Texas/75 (H3N2). In titrations of viral antigen, antibody coupled to nylon beads detected 1.9 X 10(4) plaque-forming units (PFU) per assay, whereas 2.2 X 10(5) PFU were required in assays utilizing antibody adsorbed to polystyrene beads. Use of fluorogenic or radioactive substrates for alkaline phosphatase-labeled antibodies increased the sensitivity for virus detection 10-fold with this enzyme, but were only slightly more sensitive than chromogenic substrates with peroxidase-labeled antibody.

Antigens, Viral↗