Comparison of an enzyme-linked immunosorbent assay for quantitation of rotavirus antibodies with complement fixation in an epidemiological survey

1978 ◽  
Vol 8 (3) ◽  
pp. 268-276
Author(s):  
L H Ghose ◽  
R D Schnagl ◽  
I H Holmes

The development of a micro-scale enzyme-linked immunosorbent assay (ELISA) with horseradish peroxidase as the marker enzyme for the detection and measurement of human rotavirus antibodies is described. A semipurified preparation of the serologically related simian agent, SA-11 virus, was used as the antigen. Test sera were reacted with antigen-sensitized wells in disposable poly-vinyl microplates. Any attached antibody was detected by the addition of peroxidase-labeled anti-species immunoglobulin (conjugate) followed by assay of the enzyme reaction with its substrate, hydrogen peroxide plus 5-aminosalicylic acid. This micro-ELISA was compared with complement fixation in a seroepidemiological study of the age prevalence of rotavirus antibody in Aboriginal and European populations living in the same outback area in Australia. The ELISA (results read with the naked eye) proved to be approximately 16 times more sensitive than complement fixation. Of Aborigines, 71% had rotavirus complement-fixing antibody, as compared to 45% of Europeans. By ELISA 100% of both populations had rotavirus antibodies. Mean antibody titers in the different age groups were higher in Aborigines than in Europeans. Antibody levels rose steeply throughout the first 20 years of life, remained high during the next 20 years, then increased again at least up to the age of 60 years. The micro-ELISA was practical, simple to perform, and more suitable than complement fixation for large seroepidemiological rotavirus studies. It also has potential for serodiagnosis of the disease, both in the laboratory and in the field.

2007 ◽  
Vol 15 (2) ◽  
pp. 297-302 ◽  
Author(s):  
Olga Sánchez Negrette ◽  
Fernando J. Sánchez Valdéz ◽  
Carlos D. Lacunza ◽  
María Fernanda García Bustos ◽  
María Celia Mora ◽  
...  

ABSTRACT Serological tests are the main laboratory procedures used for diagnosis during the indeterminate and chronic stages of Chagas' disease. A serological regression to negativity is the main criterion used to define parasitological cure in treated patients. The aim of this work was to monitor the individual specificities of antibody levels for 3 years posttreatment in 18 adult patients. Conventional serological techniques (hemagglutination assays and enzyme-linked immunosorbent assay [ELISA]) were modified by using recombinant antigens to detect early markers of treatment effectiveness. For this purpose, serum samples were taken before and during treatment and every 6 months after treatment for at least 3 years. When hemagglutination assays were used, a decrease in antibody levels was observed in only one patient. When ELISA with serum dilutions was used, antibody clearance became much more apparent: in 77.7% (14/18) of the patients, antibody titers became negative with time. This was observed at serum dilutions of 1/320 and occurred between the 6th and the 30th months posttreatment. The immune response and the interval for a serological regression to negativity were different for each patient. For some of the recombinant antigens, only 50% (9/18) of the patients reached the serological regression to negativity. Recombinant antigen 13 might be a good marker of treatment effectiveness, since 66.6% (six of nine) of the patients presented with an early regression to negativity for specific antibodies to this antigen (P = 0.002).


1980 ◽  
Vol 89 (3_suppl) ◽  
pp. 117-120 ◽  
Author(s):  
P. Branefors ◽  
T. Dahlberg ◽  
O. Nylén

A series of episodes of acute otitis media were studied with reference to the bacterial findings in the nasopharynx and the specific antibody response in a group of children nine months to ten years of age, with previous frequent episodes of acute otitis media, Serum IgG, IgM and IgA antibody levels against five polysaccharide antigens, namely Haemophilus influenzae type b and Streptococcus pneumoniae types 3, 6, 19 and 23, were studied by means of an enzyme-linked immunosorbent assay. The selection of polysaccharide antigens was based on isolation frequency. The sera to be tested were tenfold serially diluted. An extinction of 0.2 over the base was taken as the end-point titer and expressed as in-log10. The results showed that most children including those under three years of age showed increasing homologous antibody titers at an infection, or had already initially very high antibody titers, especially of the IgG class. The titers reached levels of 104 to 105. In some cases, however, it could be shown that high serum antibody titers did not give protection against a new infection with the same serological type of bacteria. It was also demonstrated that most children, regardless of age, had IgG and IgM titers against the heterologous antigens. In some cases the levels were quite high (103 to 104). However, the IgA antibody levels were lower and in a considerable number of samples antibodies were not even detectable.


Author(s):  
B. Davoust ◽  
T. Normand ◽  
O. Bourry ◽  
H. Dang ◽  
E. Leroy ◽  
...  

A survey of helminth parasites was carried out on 198 dogs living in almost complete liberty in villages in the northeast of Gabon. Faeces and blood samples were collected and analysed. Dirofilariaimmitis antigen was detected in 13.6 % of dogs using the SNAP 3Dx® test, a commercially available enzyme-linked immunosorbent assay (ELISA). Faecal examination revealed that 91.4 % of dogs were infected by intestinal helminths. Ascarids were found in 58.5 % of the samples. Trichuris vulpis was observed in 49.5 % of cases, and Uncinaria spp. and Ancylostoma spp. in 34.8 %, Spirocerca lupi in 25.3 % and Capillaria spp. in 10.6 %. Cestode embryophores were found in 8.6 % of the samples.


1988 ◽  
Vol 101 (2) ◽  
pp. 405-410 ◽  
Author(s):  
R. C. H Lau

SUMMARYEnzyme-linked immunosorbent assay (ELISA) tests were used to measure IgG antibody levels in 2638 New Zealand children who had been immunized with the triple vaccine DTP. The percentage of children immune to diphtheria decreased with age. The percentage of children immune to tetanus varied from 67.1 to 55.0%. The percentage of children with measurable antibody to pertussis increased with age. The mean percentages of children with measurable antibody or immunity to one or more DTP components were 34.2% (with 3 components), 34.4% (2 components), and 78.1% (1 component). It appears the immunization strategy for diphtheria and tetanus is satisfactory for herd immunity in New Zealand children. However, the current pertussis strategy may not be providing adequate immunity to 5-year-olds in this country.


1998 ◽  
Vol 5 (5) ◽  
pp. 613-616 ◽  
Author(s):  
Felix Grimm ◽  
Friedrich E. Maly ◽  
Jian Lü ◽  
Roberto Llano

ABSTRACT The potential roles of specific antibodies of the different immunoglobulin G (IgG) subclasses in the serological diagnosis of cystic echinococcosis (CE) and alveolar echinococcosis (AE) were investigated by an enzyme-linked immunosorbent assay based on hydatid fluid as antigen. Specific antibodies of subclass 1 were found to be of major importance. In sera collected at the time of diagnosis (i.e., before any therapeutic intervention was initiated) they could be demonstrated in 14 of 15 sera from patients with CE and in all 12 sera from patients with AE. The most discriminatory and the most specific antibodies found in this study belonged to IgG subclass 4. Only one false-positive reaction was observed with 253 sera from healthy volunteers, and no cross-reactions occurred in 80 sera from patients with different parasitic infections. Specific IgG4 antibodies could be demonstrated in 61.0 to 66.7% (CE) or 47.6 to 66.7% (AE) of the cases. Antibody levels of IgG subclass 2 were elevated only moderately, and subclass 3 antibodies were detected in a few cases only. In addition, nonspecific reactions in sera of healthy volunteers or patients with other parasitic infections could partially be attributed to antibodies of subclasses 2 and 3.


PEDIATRICS ◽  
1984 ◽  
Vol 74 (1) ◽  
pp. 90-93
Author(s):  
M. Dianne Murphy ◽  
Philip A. Brunell ◽  
Alan W. Lievens ◽  
Ziad M. Shehab

A measles epidemic in San Antonio, Texas provided a population of children who were immunized at ≤10 months of age and reimmunized at ≥15 months of age. Of these children, 302 were evaluated for measles antibody by the sensitive enzyme-linked immunosorbent assay (ELISA), and their responses were compared with those of 300 children who had been immunized at the customary time (≥15 months) with a single immunization. There were only five seronegative findings in each group. The children immunized at the customary time did have significantly higher (P < .001) antibody titers than the children immunized at ≤10 months and reimmunized at ≥15 months. These results indicate that early immunization followed by reimmunization may be indicated when young infants are at significant risk of measles exposure. This approach should not create an increased number of serologically nonresponsive children when reimmunized at ≥15 months.


PEDIATRICS ◽  
1988 ◽  
Vol 82 (2) ◽  
pp. 181-192
Author(s):  
John L. Sever ◽  
Jonas H. Ellenberg ◽  
Anita C. Ley ◽  
David L. Madden ◽  
David A. Fuccillo ◽  
...  

An analysis of the antibody titers to toxoplasmosis for 22,845 pregnant women in the Collaborative Perinatal Project was conducted in relation to clinical and laboratory findings in the mothers and children through 7 years of age. More than 900 observations were considered for each mother and child. The major findings were in the children and included a predicted doubling in the frequency of deafness among children born to women with antibody to toxoplasmosis, a predicted 60% increase in microcephaly, and a 30% increase in low IQ (<70) in association with the presence of high maternal antibody titer (256 to 512) to toxoplasma. A serologically defined high-risk group of mothers was identified on the basis of high indirect hemagglutination antibody levels or seroconversions and increased IgM toxoplasma antibody levels (indirect fluorescent antibody ≥32, enzyme-linked immunosorbent assay ≥0.7). Of the 15 pregnancies in this group, two children had congenital toxoplasmosis and three were stillborn.


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