The usefulness of Diffusion-In-Gel-ELISA in clinical practice as illustrated by a Campylobacter jejuni outbreak.

In this study, the DIG-ELISA (Diffusion-In-Gel Enzyme Linked Immunosorbent Assay) is presented as a tool for the determination of antibodies with improved quantitation over other solid phase assays. The method combines the diffusion of antibodies in agar with the EIA concept in Petri dishes. Diffusion of native, undiluted sera results in (1) a concentration gradient in the effort to reach equilibrium, i.e., an end point titration; (2) a separation of serum components of different sizes; (3) unlimited possibilities for migrating antibodies to bind antigenic epitopes because of the constant antigen excess. Low affinity antibodies can bind divalently and are more readily detected; and (4) elimination of dilution errors. The combination of undiluted sera and Petri dishes as solid phase also permits a large number of samples to be tested with less effort and simplifies the practical handling of EIAs, including easy coating and washing procedures, reuse of antigen and quantitation by zone areas without instrumentation. Plates can be stored for months and are available for re-examination, demonstration and transport. The whole procedure can be conducted in a closed system, i.e., when testing highly contaminated samples. The usefulness of the procedure is demonstrated by a food-borne outbreak of Campylobacter jejuni. The outbreak involved 86 persons of whom 20% were culture positive in contrast to a seropositivity of 74% with DIG-ELISA. A diffusion time of 24 h was used for diagnostic purposes. Extended diffusion times of 48 h and 72 h were utilized when consecutive series of sera showed identical values after 24 h, indicating high antibody content that resulted in a peak serum (end point). For infectious diseases with a rapid course, this assay could be used as an acute test. With the diffusion step prepared in advance, DIG-ELISA is a ready-to-use test. When frequent sampling of sera is performed in the very early phase of the disease, DIG-ELISA reveals that all Ig-classes can be present at high titer and the diagnostic potential of the immune response is better utilized. The DIG-ELISA has the methodological flexibility and the physical qualities to be an effective, inexpensive technique for quantitation of antibodies at any laboratory.

Rheumatic symptoms following an outbreak of campylobacter enteritis: a five year follow up.

Eighty six of 106 (81%) guests attending a party were followed up after an outbreak of Campylobacter jejuni enterocolitis. Acute diarrhoeal illness was reported in 35 subjects (33%), of whom seven showed acute rheumatic symptoms either alone or with other symptoms of infection with C jejuni. The antibody response to C jejuni corresponded well with the intensity of the disease. In the early phase of the gastrointestinal disease the patients with acute rheumatic symptoms displayed significantly higher IgM antibody levels in serum samples than the other patients in this study. Levels of antibodies to C jejuni were increased in serum samples from 31 patients (29%) without symptoms of infection with C jejuni. At a follow up after five and a half years, four of these patients suffered from chronic rheumatic disorders. One HLA-B27 positive woman developed reactive arthritis with a relapse seven years later. The remaining 20 subjects (19%) remained healthy and their antibody tests and stool cultures were negative for C jejuni. It is concluded that C jejuni enterocolitis is significantly associated with rheumatic symptoms in the early phase and may also cause chronic rheumatic disorders.

Toxic hepatitis following ketoconazole treatment.

Ketoconazole, a new broad-spectrum oral antifungal agent, has proved most useful in the treatment of superficial and systemic fungal infections. Chronic conditions like mucocutaneous candidiasis can be treated with but few side effects. An increasing number of cases have, however, been noted of severe toxic hepatitis, following ketoconazole treatment. A new case (a 61-yr-old woman treated for onychomycosis) is reported here.

Diffusion-in-gel enzyme-linked immunosorbent assay for routine detection of IgG and IgM antibodies to Campylobacter jejuni.

A diagnostic serologic test for antibodies to Campylobacter jejuni is presented. A diffusion-in-gel enzyme-linked immunosorbent assay with a surface antigen pool from two C jejuni strains was used. In testing serum samples for antibodies to C jejuni this system possessed high specificity, reliability, sensitivity, and capacity. It is easy to perform and there is no demand for expensive equipment. IgG and IgM antibodies can be quantitated separately, which allows for discrimination between individuals with acute infections and those with antibodies remaining after recovery. One serum sample is often sufficient to obtain such information. Seven groups of individuals were investigated. Serum samples from a total of 159 patients in three different groups and from 306 healthy individuals in four different groups were tested.

Epidemiological aspects of Campylobacter jejuni enteritis.

An epidemiological study on Campylobacter jejuni enterocolitis was performed in an urban Swedish community. The study included 55 patients gathered during a six-month period. Forty-one of the 55 patients (75%) were infected outside Sweden. Campylobacter enterocolitis was rare among children within the country. Patients infected in Sweden had eaten chicken significantly more often than a corresponding control group. Seven out of nine chicken consuming campylobacter patients also had prepared the fresh chicken alone, and none of their family members became ill. Thus the preparation of food contaminated with Campylobacter seems to elevate the risk for contracting the disease. Sick household pets transmitted the campylobacter infection to two patients. Forty-six of the patients had a total of 85 close household members. Three definite secondary cases were found. There was no evidence of transmission of Campylobacter by food prepared by two cooks who were working while still being asymptomatic excreters. Clinical reinfection with Campylobacter was observed in one patient. No patients became long-term carriers of Campylobacter.

Intestinal occurrence of Campylobacter fetus subspecies jejuni and Clostridium difficile in children in Sweden.

Stool samples were cultured from 356 children in different states of health and in different age groups between birth and six years of age in order to investigate the occurrence of Campylobacter jejuni and Clostridium difficile. Campylobacter jejuni was isolated from two of 56 children with diarrhoea but was not isolated from any of 300 healthy children or children recently treated with antibiotics. Campylobacter jejuni does not seem to be a common cause of diarrhoea in children in Sweden and is definitely not a member of the normal fecal flora. Clostridium difficile was isolated from 17 of 100 healthy children one week to one year old, and from one of 100 perinatal children. About the same isolation frequency was found in children with diarrhoea and in children recently treated with antibiotics. A total of 34 isolates of Clostridium difficile were obtained, 29 of which were from children less than one year old. The isolation of Clostridium difficile in stools of children should in most cases be considered a normal finding.

The occurrence of Campylobacter jejuni in fresh food and survival under different conditions.

Campylobacter jejuni was an almost regular finding in chickens and in minced meat from pigs and cattle sold in ordinary food stores. The bacteria survived on the food at 4 degrees C for one week and frozen at -20 degrees C for three months. None of the strains tested survived heat treatment at 60 degrees C for longer than 15 min. C. jejuni is apparently a frequent guest in most kitchens. Correct food handling and heat treatment to at least 60 degrees C for 15 min should be enough to prevent infection.

Clostridium difficile in relation to enteric bacterial pathogens.

All feces samples (n = 2,390) sent to the Bacteriological Laboratory, Göteborg, Sweden over 43 days were, in addition to the standard procedure, cultivated to detect Clostridium difficile by using a special selective medium. C. difficile was found in 81 of the 2,390 samples (3%). These 81 samples represented 56 patients. Fifty of the 56 patients had diarrhea. In 20 of the 56 patients (36%), Salmonella, Campylobacter, or Yersinia were also found. Of the 2,390 samples 252 (11%) from 132 patients revealed positive isolations of Salmonella, Shigella, Campylobacter, or Yersinia in comparison to 3% for C. difficile alone. This result suggests that C. difficile can easily be isolated with proper techniques. Concomitant isolations of more than one bacterial pathogen in cases of gastroenteritis were often found for C. difficile. The theory presented here is that any change of the normal bacterial fecal flora due to such causes as antimicrobial treatment or enteric infections like Salmonella increases the possibilities of isolating C. difficile. The causative significance of C. difficile might in most cases be doubtful. The majority of cases with diarrhea and C. difficile were self-healing and not severe. Only 2 cases of 56 had severe diarrhea with extended engagement of the colonic mucous membrane, but with no signs of pseudomembranes.

Campylobacter fetus subspecies jejuni: a common cause of diarrhea in Sweden.

Stool samples from approximately 2,550 patients with gastrointestinal infections were cultured for bacterial pathogens between January 1978 and September 1979, and 277 campylobacter (10.9%), 183 salmonella, 89 shigella, and 17 yersinia infections were identified. Campylobacter was found in all age groups, with the majority in the age group of 20-34 years. Most cases of campylobacter diarrhea were isolated during summer or late fall. After five weeks, 90% of the patients had no Campylobacter in their stools. The incubation time was estimated to be one to six days. Campylobacter gastroenteritis, even more common than salmonella gastroenteritis, was in general not regarded as a serious disease, despite the fact that most patients had marked symptoms. Almost all patients recovered without antibiotic treatment, although in some cases the disease was long-lasting, and 50 of the 277 patients required hospitalization. Thus, Campylobacter must be considered in the diagnosis of patients with diarrhea.

Phlebitis induced by parenteral treatment with flucloxacillin and cloxacillin: a double-blind study.

Two studies were performed on a total of 54 patients with staphylococcal infections. Study I compares with phlebitogenic properties of flucloxacillin after intravenous infusions when either saline or sterile water was used as a solvent. No difference was observed between the two solvents, and the frequency of phlebitis for the total material without respect to solvents was 5% after 1 day of treatment and 13% after 2 days. Study II was a double-blind comparison of phlebitis caused by intravenous infusions of either flucloxacillin or cloxacillin. The frequencies of phlebitis were found to be 18 and 13%, respectively. After 2 days of treatment the frequency of phlebitis increased dramatically for both drugs. All infusions were given through a plastic cannula of 5-cm length and 1.2-mm diameter.

Treatment of toxoplasmosis with trimethoprim-sulphamethoxazole.

Toxoplasmosis is known to complicate diseases with impaired cellular immunity. For treatment of toxoplasma infections atoxic drugs should be used to avoid depression of the bone marrow. The hitherto recommended treatment, pyrimethamine in combination with sulphonamides, is often associated with severe side-effects. The present study presents the results of treatment with trimethoprim and sulphamethoxazole in 7 patients, clinically and serologically diagnosed as having toxoplasmosis. Good therapeutic results were observed in 5 patients with lymphoglandular toxoplasmosis and a significant reduction of the dye test titres were found in 6 patients. In one patient, however, a relapse of clinical symptoms and a reversion to high dye test titres were observed 6 1/2 months after the end of the treatment. Treatment had to be discontinued in one patient due to an allergic reaction.