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At least 163 records · Page 9Linked to original sources
PubMed1996

Multi-well ELISA based on independent peptide antigens for antibody capture. Application to Lyme disease serodiagnosis.

Novel procedures for the use of peptides as antibody-capture reagents in the ELISA format have been investigated. Epitope sequences from known immunodominant antigens of Borrelia burgdorferi were selected by screening peptide libraries with sera from patients with Lyme disease. Several epitope peptides were synthesized and immobilized, separately, on the ELISA plate as haptens on bovine serum albumin. Based on a comparative analysis of serum samples, it appears that peptide antigens can be used as effectively as a whole cell lysate to discriminate between Lyme disease and non-Lyme disease sera, thus avoiding dependence on bacterial sonicates which vary from passage to passage. Further improvements in epitope design for enhancement of accuracy in serodiagnosis are discussed.

Amino Acid Sequence↗
PubMed1981

Serodiagnosis of human Schistosoma mansoni infections: enhanced sensitivity and specificity in ELISA using a fraction containing S. mansoni egg antigens omega 1 and alpha 1.

Six fractions of Schistosoma mansoni egg homogenate obtained by cation exchange chromatography were tested in an enzyme-linked immunosorbent assay (ELISA) for their efficacy in serodiagnosis of human S. mansoni infections. One cationic fraction (CEF6) containing egg antigens omega 1 and alpha 1, was found to be highly reactive with a S. mansoni positive reference serum pool. 94 of 95 sera from individuals infected with S. mansoni gave a positive reaction with CEF6, and no cross reactivity was obtained with this antigenic fraction and sera from donors infected with heterologous non-schistosome parasites and cases of avian cercarial dermatitis. Only 10 of 165 sera from patients with S. haematobium and three of 10 sera from patients with S. japonicum were positive with S. mansoni egg CEF6. In one group of patients from West Africa, examined both serologically and parasitologically at a six-monthly interval, the measurement of antibody to CEF6 provided a more sensitive diagnostic aid than stool examination.

Animals↗
PubMed1981

Serodiagnosis of human strongyloidiasis by an enzyme-linked immunosorbent assay.

The sensitivity specificity of an enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of strongyloidiasis has been investigated. 45 men with long-standing strongyloidiasis were compared with the same number of age- and sex-matched control subjects. The ELISA detected antibody in 84% of patients with parasitologically proven strongyloidiasis. When the technique was compared with an indirect immunofluorescent assay (IFA), a high correlation coefficient was obtained. Specificity was demonstrated by observing a marked fall in optical density of pooled positive serum after prior incubation with Strongyloides ratti soluble antigen but not after incubation with antigens derived from Ascaris suum or Dirofilaria immitis. The test is simple and offers a useful method for the diagnosis of strongyloidiasis. In these patients it was more reliable than a single parasitological examination of faeces or duodenal contents.

Antibody Specificity↗
PubMed1983

Evaluation of a visually read ELISA for serodiagnosis and sero-epidemiological studies of kala-azar in the Baringo District, Kenya.

ELISA for detecting leishmanial antibodies was established at Baringo District Hospital Kabarnet, Kenya, and successfully applied in clinical routine and sero-epidemiological surveys. To simplify the procedure and to make it more economical, ELISA was read as visual endpoint titre and blood was collected by the blotting paper method. 63 parasitologically confirmed kala-azar patients all showed a diagnostic IgG specific ELISA titre of 1:27,000 or more, which indicates a sensitivity of 100%. 11 of 14 persons found with this diagnostic titre during a house-to-house survey in a known focus of kala-azar could be confirmed parasitologically. Two more showed typical signs and symptoms of kala-azar and were successfully treated accordingly. Controls from unaffected areas were all negative. Low titres of 1:1000 and 1:3000 were observed in healthy individuals from foci of kala-azar, but could be easily distinguished from active cases. ELISA has been shown to be a convenient and reliable method for the serodiagnosis of East African kala-azar. The technique can be integrated into the routine work of a clinical laboratory at the level of a district hospital, without sophisticated equipment and with a minimum of funds. Blood of suspected kala-azar cases can be collected on blotting paper in dispensaries and health centres near known foci and sent to the laboratory.

Antibodies↗
PubMed1987

Serodiagnosis of typhoid fever in paediatric patients by anti-LPS ELISA.

Enzyme-linked immunosorbent assay (ELISA) and immunoblot analysis showed that up to 50% of the anti-typhoid antibody in sera from blood culture positive paediatric typhoid fever patients is directed against lipopolysaccharide (LPS) antigen. Anti-Salmonella typhi LPS ELISA was therefore compared to Widal agglutination for serodiagnosis of typhoid fever in paediatric patients. Sera from 38 paediatric control individuals were ELISA negative for anti-S. typhi LPS IgG; all but 2 of these specimens were negative for anti-S. typhi LPS IgM. Paediatric patients hospitalized with signs and symptoms of typhoid fever were separated into 4 groups and tested by ELISA with the following results: 46 patients negative by both culture and Widal agglutination tests, 48% positive for anti-S. typhi LPS IgG and 35% for anti-S. typhi LPS IgM; 22 negative by culture but with positive Widal titres, 82% and 68% positive respectively; 28 culture positive for S. typhi, 93% and 82% respectively; and 12 culture positive for Salmonella other than S. typhi, 92% and 92% respectively. These data suggest that anti-S. typhi LPS ELISA is a suitable assay for diagnosis of typhoid fever in children.

Adolescent↗
PubMed1987

Use of phenolic glycolipid 1 for serodiagnosis of leprosy in a high prevalence village in Papua New Guinea.

This study reports on the usefulness of an IgM phenolic glycolipid 1 (PGL-1) ELISA for serodiagnosis of leprosy in the first year of a prospective longitudinal community survey in a high (8.8%) prevalence village in Papua New Guinea. The IgM PGL-1 ELISA had limited value as a screening method for detection of new cases. Many normal persons, particularly children, had elevated IgM anti-PGL-1 antibodies, presumably a consequence of early subclinical infection.

Antigens, Bacterial↗
PubMed1990

Detection of anti-listeriolysin O for serodiagnosis of human listeriosis.

To see whether detection of antibodies against listeriolysin O (LLO) could be used to diagnose human listeriosis, sera from 28 patients infected with Listeria monocytogenes and 101 controls were tested by dot-blot titration with purified LLO. 27 patients (96.4%) with listeriosis produced specific anti-LLO. Anti-LLO was detected in 8 (15.6%) of 51 healthy controls and in 6 (12.0%) of 50 controls who had various bacterial, fungal, and viral infections. Anti-LLO titres did not exceed 100 in these two control groups. Anti-LLO could be detected soon after clinical onset of listeriosis, and antibodies persisted for at least several months. This test might be useful for epidemiological surveys and for serodiagnosis of listeriosis, especially when bacteria cannot be isolated.

Adolescent↗
PubMed1979

Bovine adenoviruses--IV. Two mixed antigens for routine serodiagnosis by complement fixation reaction.

The frequency of bovine adenovirus infections occurring in cattle has been grossly underestimated up to the present day. Primary isolation of serotypes belonging to bovine subgroup II is cumbersome. Hardly any laboratory has used all 8 officially-recognised bovine serotypes in seroneutralisation-tests, which should be done when this type-specific method is used for serodiagnosis. The complement fixation test, known as group-reactive from human adenovirus serology, has failed to disclose the true incidence of bovine infections, as until recently the importance of a novel additional group-reactive bovine adenovirus antigen has been undisclosed. Here we describe production, composition and performance of two mixed antigens for complement fixation reactions, which take into account recent findings by our team on peculiarities of bovine adenovirus antigens. Mixed antigen 1 contains bovine serotypes 1, 2 and 3 and detects group-specific antibodies of the classical mastadenoviruses. Mixed antigen 2 contains bovine serotypes 4, 5, 6, 7 and 8 and determines group-specific antibodies against the novel paramastadenoviruses. In addition, each antigen is capable of demonstrating complement-fixing type-specific antibodies in sera against the respective types included in each mixed antigen, with a net enhancing effect produced by mixing. Use of the 2 mixed antigens promptly shows whether bovine adenoviruses, presently recognised as well as unclassified, are involved in a given outbreak of respiratory or enteric disease of cattle. If needed, the responsible type may be determined in a second step of serological investigation.

Adenoviridae↗
PubMed1992

Assessment of assays for the serodiagnosis of Venezuelan equine encephalitis.

An enzyme-linked immunosorbent assay (ELISA), an immunofluorescence assay (IFA), a plaque-reduction neutralization (PRN) assay and an immunoblot assay, all by means of an antigen prepared from the attenuated Venezuelan equine encephalitis (VEE) vaccine strain of virus, were compared with the conventional haemagglutination-inhibition (HAI) assay for the serodiagnosis of VEE. The HAI assay, which includes the use of wild type virus antigen, was less sensitive than the other assays when known-positive samples of serum from an epidemic of VEE were tested. The superior sensitivity of the IgG ELISA was confirmed by assaying both VEE epidemic samples and a bank of samples from VEE vaccinees. Samples with antibody specific for other Alphaviruses, however, cross reacted weakly in this assay. The PRN, immunoblot and HAI assays, although less sensitive than the ELISA, proved more specific. Experimental infection of guinea-pigs demonstrated the value of the IgM ELISA in the early detection of VEE virus infection. Immunoglobulin M was first found at 4 days post-inoculation (p.i.) during the viraemic phase of infection. Immunoglobulin G was detected by ELISA, PRN assay and IFA at 6 days p.i. Immunoblot and HAI assays, however, did not give positive results until 10 days p.i. The results support the diagnostic use of ELISA for detecting VEE virus-specific IgM and IgG, and the use of the specific PRN assay for confirming the diagnosis.

Animals↗
PubMed1996

IgM-specific serodiagnosis of acute human cytomegalovirus infection using recombinant autologous fusion proteins.

Portions of three human cytomegalovirus (HCMV) polypeptides, which were shown previously to be highly reactive with patient sera, were expressed in Escherichia coli as autologous fusion proteins. Purified recombinant polypeptides were used as antigens in enzyme linked immunosorbent assay (ELISA) and compared against assays which use natural viral antigen from cell culture for their ability to improve IgM-specific serology of acute HCMV-infection. A fusion protein (CM2) which contained two copies of the C-terminal portion of pUL44 (p52, aa 297-433) and one copy of a highly reactive fragment of the major DNA-binding protein pUL57 (aa 545-601) proved to be superior in sensitivity and specificity compared to assays which use culture derived antigen. A construct expressing one copy of the fragments from pUL44 and pUL57 in fusion with the 54 amino terminal residues of pUL32 (pp150, aa 994-1048) did not lead to an improved sensitivity compared to CM2. Adversely, this polypeptide reacted with a number of sera from asymptomatic blood donors infected latently with HCMV indicating low specificity of this antigen for the detection of acute infection. Concordant results were obtained with an antigen that combined only the C-terminal portions of pUL44 and pUL32 (CM3). ELISA experiments with sequential sera from renal transplant recipients demonstrated that detection of IgM-antibodies using CM2 as antigen correlated closely with acute infection, whereas high levels of IgM-antibodies against CM1 and CM3 persisted for a month following acute HCMV-infection. These results indicate that the application of a single autologous fusion protein like CM2 as antigen for recombinant ELISAs can improve significantly IgM-serodiagnosis of acute HCMV infection.

Antibodies, Viral↗
PubMed1986

Species-specific serodiagnosis of equine piroplasma infections by means of complement fixation test (CFT), immunofluorescence (IIF), and enzyme-linked immunosorbent assay (ELISA).

The increasing horse trade requires a reliable immunodiagnosis of equine piroplasma infections due to import restrictions imposed by various countries, including the United States of America. It was the aim of our investigations to establish the suitability of serological tests for the detection of parasite carriers and, eventually, to differentiate between Babesia caballi and B. equi infections. The investigations were carried out on 11 ponies with experimentally-induced B. caballi and/or B. equi infection. The infections were confirmed by the demonstration of parasites in blood smears 2-13 days post infection (PI). The complement fixation test (CFT), the indirect immunofluorescence (IIF) and the enzyme-linked immunosorbent assay (ELISA) were employed for the demonstration of antibodies, and different antigen preparations were tested for their suitability. Antibodies could be demonstrated by all three tests. Complement-fixing antibodies disappear after 2-3 months PI in B. caballi-infected horses, while the IIF and ELISA gave positive results during latent infection. A reliable serodiagnosis thus requires the use of the CFT and IIF, since parasite carriers may appear seronegative by the CFT. Serological differentiation between B. caballi and B. equi was possible by CFT and, to a certain extent, by IIF during early infection, but not by ELISA. The successful treatment of B. caballi infections with Berenil could only be confirmed serologically by IIF.

Animals↗
PubMed1989

Characterization and purification of Parafilaria bovicola antigens by chromatofocusing to enhance specificity in serodiagnosis.

A study was conducted to determine if the purification of Parafilaria bovicola antigens can increase the specificity of serodiagnosis of parafilariasis in enzyme-linked immunosorbent assay. Antigens released from adult worms of P. bovicola were separated by chromatofocusing on a polybuffer exchanger of the pH range 7.3-4.0 Polypeptide analysis by sodium dodecyl sulphate polyacrylamide gel electrophoresis showed the presence of four major polypeptides with MWs of 41, 36, 24 and 20 kDa. Additional biochemical characterization identified the 24- and 20-kDa polypeptides as hydrophobic glycoproteins. The chromatofocusing purification procedures were also applied for separation of a whole-worm extract. Again, the 41- and 36-kDa antigens were identified in separate peak fractions. Using ELISA, it was shown that the 41- and 35-kDa antigens were recognized by bovine antibodies specific for P. bovicola, but not by other sera collected from cattle infected by Onchocerca gutturosa, Onchocerca lienalis, Ostertagia ostertagi and Dictyocaulus viviparus. The serological evaluation strongly suggests that the 41- and 36-kDa antigens are P. bovicola specific.

Animals↗
PubMed1996

Use of excretory/secretory antigens for the serodiagnosis of Anoplocephala perfoliata cestodosis.

Whole worm extract (WWE) and excretory/secretory (E/S) antigens of Anoplocephala perfoliata were characterised by SDS-PAGE and their use in the serodiagnosis of equine cestodosis was evaluated. An enzyme-linked immunosorbent assay (ELISA) was used to compare WWE and E/S antigen as the capture layer in an antibody capture ELISA. E/S antigen gave the best differentiation between sera from tapeworm-positive and tapeworm-negative horses. The E/S-ELISA was optimised and validated against sera from horses of known tapeworm status. This assay gave a diagnostic sensitivity of 68% (n = 38) and a specificity in helminth-naive horses of 95% (n = 20). Western blot analysis of tapeworm-positive and tapeworm-negative sera revealed the immunoreactive proteins of E/S antigen to be those of 12 and 13 kDa.

Animals↗
PubMed1996

Evaluation of dot enzyme-linked immunosorbent assay (dot-ELISA) for the serodiagnosis of canine leishmaniosis as compared with indirect immunofluorescence assay.

A dot enzyme-linked immunosorbent assay (dot-ELISA) was developed and compared with a standard indirect immunofluorescence assay (IFA) for the rapid serodiagnosis of canine leishmaniosis. The two tests were used to examine sera from Leishmania infantum-infected and control dogs. Using the dot-ELISA, 137 of 149 sera (91.9%) from infected animals gave a clearly positive reaction, whereas in the standard IFA technique 147 (98.7%) were positive at a reciprocal titer of 40 or over (titer range 40-10,240). Control sera from 75 healthy dogs, not living in endemic areas, and 11 dogs with other diseases (babesiosis, cryptococcosis, ehrlichiosis, dermatitis, and chronic hepatitis) but Leishmania-negative were used to determine the specificity of the assays. All the sera were negative with IFA (100%), whereas using the dot-ELISA only 74 sera (86%) from controls gave a negative result. In the standard IFA no cross-reactivity was noted, in the dot-ELISA a weak cross-reaction was observed with a serum sample from a dog with babesiosis. The interpretation of dot-ELISA could be easily performed by visual inspection of the nitrocellulose disks. The most remarkable feature of dot-ELISA was the high sensitivity (91.9%) and positive predictive value (92.6%), even if the test showed a specificity lower than IFA. Because of its easy performance and high sensitivity, the dot-ELISA may be a useful test to be executed in the field for the diagnosis of canine leishmaniosis.

Animals↗
PubMed1994

Evaluation of a saline boiled extract, capsular polysaccharides and long-chain lipopolysaccharides of Actinobacillus pleuropneumoniae serotype 1 as antigens for the serodiagnosis of swine pleuropneumonia.

A saline boiled extract (SBE), capsular polysaccharides (CPS) and long-chain lipopolysaccharides (LC-LPS) of Actinobacillus pleuropneumoniae serotype 1 have been evaluated in ELISA for the serodiagnosis of swine pleuropneumonia caused by this serotype. Mean optical densities (ODs) obtained with the three antigens using sera from negative herds as well as from animals experimentally and naturally exposed to A. pleuropneumoniae serotypes 1, 9 or 11 were not significantly different. The positive ELISA reaction with anti-serotypes 9 and 11 was unexpected with the CPS, which are supposed to be serotype-specific; LPS, and to a lesser extent proteins, were present in the CPS and appeared to be responsible for this reaction. In addition, sera from animals exposed to a field strain of A. pleuropneumoniae serotype 3 and to Actinobacillus suis presented a significantly lower mean OD (P < 0.001) when LC-LPS were used. Cross-reacting antigens consisted mainly of LPS core-lipid A present in the SBE and CPS. The specificity and the sensitivity of the ELISA were evaluated using three different cut-off values (the OD plus two, three and four times the standard deviation or SD) obtained with 667 negative sera. The diagnostic sensitivity was of 81% with the three antigens and the different thresholds. The diagnostic specificity was of 84, 86 and 88% for the mean plus two, three and four times the SD respectively using the SBE and the CPS, while that obtained with the LC-LPS was of 96, 98 and 99% using the same thresholds. In conclusion, LC-LPS make an easily obtainable antigen and seem to retain the best specificity while minimizing losses of sensitivity.

Actinobacillus Infections↗
PubMed1994

Evaluation of long chain lipopolysaccharides (LC-LPS) of Actinobacillus pleuropneumoniae serotype 5 for the serodiagnosis of swine pleuropneumonia.

Long chain lipopolysaccharides (LC-LPS) of Actinobacillus pleuropneumoniae serotype 5 have been evaluated and compared with a crude boiled extract (CBE) in ELISA for the serodiagnosis of swine pleuropneumonia caused by this serotype. The mean optical density (OD) obtained with the LC-LPS in ELISA using sera from negative herds as well as from animals experimentally and naturally exposed to A. pleuropneumoniae serotype 5 was not significantly different from that obtained with the CBE. However, sera from animals exposed to serotypes of A. pleuropneumoniae other than serotype 5 presented a significantly lower mean OD (P < 0.05) when the LC-LPS was used. As a consequence, it was demonstrated that a high percentage of non-specific cross-reactions were eliminated, without losing specificity. The specificity and the sensitivity of the LC-LPS- and CBE-ELISA were evaluated using two different cut-off values (the OD plus two and three standard deviations) (SD) obtained from 593 sera from negative herds. The LC-LPS appeared a more suitable antigen than the CBE, since the sensitivity and the specificity (obtained with both thresholds) were statistically improved (P < 0.01). A threshold of 0.244 (mean OD plus three SD) for the LC-LPS-ELISA seemed more suitable, since a sensitivity of 79% and a specificity of 97% was achieved. Nevertheless, it may be advisable to keep a buffer range (OD between 0.194 and 0.243) and to consider sera presenting values within this range as suspicious. In the present study, the complement fixation test presented a high specificity (97%) and a very low sensitivity (47%). A herd with animals presenting ELISA positive and CFT negative results in serology, along with the absence of suggestive lesions should not be considered as a non-infected herd.

Actinobacillus Infections↗
PubMedDate not supplied

Evaluation of the use of 5-mycoloyl-beta-arabinofuranosyl-(1-->2)-5-mycoloyl- alpha-arabinofuranosyl-(1-->1')-glycerol in serodiagnosis of Mycobacterium avium-intracellulare complex infection.

5-Mycoloyl-beta-arabinofuranosyl-(1-->2)-5-mycoloyl-alpha-ar abinofuranosyl-(1-->1')-glycerol, an antigenic glycolipid from the Mycobacterium avium-intracellulare complex (MAC) was examined for its applicability to the serodiagnosis of MAC infection by ELISA. Serum IgM antibody titres against this glycolipid in healthy controls, pulmonary tuberculosis patients and sputum-MAC-culture-negative MAC patients were generally below the cut-off point (ELISA-negative), whereas most of the MAC-culture-positive MAC patient sera were ELISA-positive (93.5%) and their titres were often very high. Thus, high serum IgM titres against this glycolipid may be said to imply that the MAC disease is in an active phase.

Adult↗
PubMed1994

Screening of synthetic trehalose 6,6'-diesters and trehalose 6-monoesters as potential immunoreactants for the serodiagnosis of tuberculosis.

The absence of serological cross-reactivity between trehalose 2,3-diester (DAT, formerly SL-IV) and synthetic trehalose 6,6'-diesters and trehalose 6-monoesters was established by ELISA testing using polyclonal immune sera raised in rabbits sensitized with "DAT". From the screening of fifteen synthetic trehalose 6,6'- and 6-esters, "mirror" pseudo cord factor no. 1, "mirror" amides no. 5 and 6, cord factor analogues 7 and 8 and trehalose 6-monoesters 10 and 11 were selected for future, more extensive serological analysis. Paired comparisons of analogues among these fifteen substances showed that serodiagnostic discrimination power was more a function of the carbon chain length of their substituent groups--as well as of their position--than of the "mirror" constitution of the molecules. More exhaustive testing of these seven compounds is needed to select the synthetic product most efficient in the ELISA serodiagnosis of tuberculosis.

Animals↗