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PubMed1999

Evaluation of FAST-ELISA for the diagnosis of experimental trichniosis.

In order to compare between FAST-ELISA and ELISA for the diagnosis of experimental trichinosis and study the kinetics of antibody and eosinophilic responses, six New Zeland rabbits were infected orally by Trichinella spiralis larvae. Blood was collected every other day for the first 2 weeks, then weekly for eleven weeks post infection. T. spiralis crude larval antigen was prepared for coating of ELISA plates and FAST-ELISA beads. Blood was examined for eosinophilic count and for serum antibody level by ELISA and FAST-ELISA techniques. The burden of infection was assessed by counting encysted larvae in muscle samples of the infected rabbits. By FAST-ELISA antibodies were detected seven days post infection (P.I.), while with ELISA technique antibodies were detected after 10 days. Both tests detected maximum antibody levels on the 4th week. The eosinophilic count reached its peak by the 2nd week. There was a significant inverse correlation between the mean eosinophilic count and the mean larval count. FAST-ELISA proved to be more sensitive than ELISA in early detection of infection, besides being a simple, fast and sensitive assay for antibody detection against T. spiralis.

Animals↗
PubMed1986

Clinical significance of enhanced sensitivity in detecting HBsAg by Biotin-Avidin ELISA.

The Biotin-Avidin System (BAS) was incorporated into enzyme-linked immunosorbent assay (ELISA) technique to increase the sensitivity of the standard ELISA for the detection of HBsAg. We compared the sensitivity of Biotin-Avidin ELISA (BA-ELISA) with monoclonal antibody reverse passive hemagglutination assay (McAB RPHA), standard ELISA and solid phase radioimmunoassay (SPRIA). BA-ELISA was most sensitive. Out of 276 healthy individuals who were HBsAg negative by polyclonal antibody RPHA (PcAB-RPHA), 10(3.60%) were HBsAg positive by the BA-ELISA. Out of 123 acute hepatitis patients who were HBsAg negative by PcAb-RPHA and anti-HBc positive by ELISA, 64(52.03%) were found to be HBsAg positive by BA-ELISA. This study indicates that enhanced sensitivity of BA-ELISA in detecting HBsAg significantly increases the correctness of diagnosis of HBV infection.

Avidin↗
PubMed1983

Sensitivity and specificity of Czechoslovak ELISA HBsAg Sevac tests: comparison with other third-generation tests and counter-immunoelectrophoresis.

Two variants of sandwich-type ELISA (Enzyme Linked Immunosorbent Assay) kits for HBsAg detection (Sevatest ELISA HBsAg Macro I and Sevatest ELISA HBsAg Micro I) in human sera and plasmas were developed. As the solid phase, the ELISA Macro kit and ELISA Micro kit make use of polystyrene microtubes, and polystyrene microtitration plates, respectively, of Czechoslovak production (Koh-i-noor, Dalecín). Capture anti HBs antibody for adsorption to solid phase and rabbit anti HBs antibody for labelling with horse-radish peroxidase were prepared for both tests. The sensitivity of both ELISA kits for HBsAg, equal to approx. 2 ng/ml, was determined by titrating six selected HBsAg-positive sera and the WHO Agk 76 panel of HBsAg-positive sera and the results were compared with those obtained by ELISA, RIA (Radioimmunoassay) and RPHA (Reverse passive hemagglutination) kits of different producers and by counter-immunoelectrophoresis (CIEP). The sensitivity of the new ELISA kits was comparable to that of other producers' ELISA kits, higher than that of RPHA kits and only a little lower than that of RIA kits. A set of sera of patients hospitalised with different diagnoses was tested for HBsAg. The detection rate by ELISA Macro kit 2.8 and 1.5 times higher than by CIEP and RPHA (Raphadex B), respectively, and 1.1 time lower than by RIA (Austria II).

Counterimmunoelectrophoresis↗
PubMed1993

Hepatitis C infection by polymerase chain reaction in alcoholics: false-positive ELISA results and the influence of infection on a clinical prognostic score.

Antibody to hepatitis C as measured by the ELISA method is common in alcoholics. The presence of antibody to C 100-3 has been associated with more advanced disease. However, few studies have investigated the clinical significance of hepatitis C infection as defined by the presence of circulating viral RNA in alcoholics. We have prospectively examined 48 consecutive alcoholic patients for the presence of antibody to hepatitis C by an ELISA for antibody to the C100-3 antigen and by the reverse transcriptase polymerase chain reaction (PCR) using nested primers for the 5' nontranslated region of the viral RNA. Patients with liver disease were scored for disease severity by the combined clinical and laboratory index (CCLI). Overall, 12 of 48 patients (25%) were ELISA positive and eight of 48 (16%) were PCR positive. Among the 34 patients with liver disease, 10 (29%) were ELISA positive and six (18%) were PCR positive. All PCR-positive patients were also ELISA positive. There was no significant difference in the disease severity score (CCLI) or the duration of clinical disease in PCR-positive versus PCR-negative patients with liver disease. However, PCR-positive patients were significantly younger (43 +/- 6 vs. 55 +/- 10 yr, p = 0.001), indicating an earlier onset of severe disease in PCR-positive patients. There were no false-negative ELISA tests in either those with or those without liver disease. Among the 34 patients with liver disease, four of 10 patients with positive antibody were negative by PCR. Neither individual immunoglobulin levels (IgG, IgM, IgA) nor total globulins were significantly different between the ELISA-positive/PCR-negative patients and ELISA-positive/PCR-positive patients. When the entire group of 34 patients with liver disease was considered, we could not detect a significant correlation between ELISA absorbance and total globulins, and only a weak correlation between absorbance and immunoglobulin G (p = 0.49). These data show that the majority of alcoholic patients with liver disease and positive antibody to hepatitis C also have demonstrable viremia by PCR, and may require further evaluation and treatment. Elevated immunoglobulins in these patients do not correlate strongly with ELISA absorbance for anti-HCV. The presence of clinically advanced disease at a significantly younger age in the PCR-positive group is consistent with the concept of synergy between active viral infection and alcohol abuse in the development of liver disease in alcoholic patients.

Alcoholism↗
PubMed2000

Test characteristics of immunofluorescence and ELISA tests in 856 consecutive patients with possible ANCA-associated conditions.

OBJECTIVE: To examine the test characteristics of immunofluorescence (IF) and enzyme-linked immunosorbent assays (ELISA) in a consecutive series of patients under evaluation for anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV). METHODS: Using stored sera, we performed a cross-sectional study on 856 consecutive patients tested prospectively for ANCA by IF, Based on guidelines from the 1994 Chapel Hill Consensus Conference (CHCC), we determined each patient's underlying diagnosis by a medical records review without regard to their ANCA status (the CHCC guidelines do not require ANCA as a prerequisite for diagnosis). We grouped patients with forms of vasculitis commonly associated with ANCA into one of 4 types of AAV: Wegener's granulomatosis (n = 45), microscopic polyangiitis (n = 12), Churg-Strauss syndrome (n = 4), and pauci-immune glomerulonephritis (n = 8). We also classified patients without clinical evidence of AAV (92% of all patients tested) into 5 predefined categories of disease (including "other") and an additional category for no identifiable disease. In a blinded fashion, we then performed ELISAs on the stored serum for antibodies to proteinase-3 (PR3) and myeloperoxidase (MPO) and calculated the test characteristics for both ANCA assay techniques. RESULTS: Sixty-nine of the 856 patients (8.1%) had clinical diagnoses of AAV based on CHCC guidelines. The positive predictive value (PPV) of ELISA for AAV was superior to that of IF, 83% versus 45%. For patients with both positive IF tests and positive ELISA tests, the PPV increased to 88%. Both IF and ELISA had high negative predictive values (97% and 96%, respectively). Positive ELISA tests were associated with higher likelihood ratios (LR) than IF (54.2 [95% CI = 26.3, 111.5] versus 9.4 [95% CI = 6.9, 12.7]). The LR of both a positive IF and a positive ELISA was 82.1 (95% CI = 33.3, 202.5). CONCLUSIONS: Compared with IF, an ELISA test fo ANCA was associated with a substantially higher PPV and LR for AAV. This fact, combined with the greater sensitivity of IF, suggests that an effective testing strategy is to perform ELISA tests only on samples that are positive for ANCA by IF.

Antibodies, Antineutrophil Cytoplasmic↗
PubMed1993

Antibodies to Sm, RNP and SSB detected by solid-phase ELISAs using recombinant antigens: a comparison study with counter immunoelectrophoresis and immunoblotting.

Sera from 64 patients with systemic lupus erythematosus (SLE) were tested for the presence of anti-Sm, anti-RNP, and anti-SSB antibodies using commercially available solid-phase enzyme-linked immunosorbent assays (ELISAs) and recombinant nuclear proteins as substrates. The results were compared to those obtained with counterimmunoelectrophoresis (CIE) and immunoblotting (IBT) using a rabbit thymus extract (RTE) as the substrate. The ELISAs detected antibodies to Sm, RNP, and SSB in, respectively, 25%, 36%, and 15% of the SLE sera. Neither IBT-positive/ELISA-negative nor CIE-positive/ELISA-negative sera were found, regardless of the specificity considered, suggesting that ELISAs using recombinant nuclear antigens are highly sensitive. Discrepancies were observed between the results obtained with these different techniques. In addition to sera positive by both IBT and ELISA but negative by CIE, a substantial number of sera had ELISA-detectable anti-Sm, anti-RNP, and anti-SSB antibodies which failed to react with the corresponding polypeptides by IBT. The reasons for ELISA/IBT discrepancies were explored; however, no single explanation was found. Instead, a higher sensitivity of the ELISA to detect antibodies directed against certain polypeptides, the possible inability of IBT using RTE as the substrate to detect antibodies reacting with conformational antigenic determinants, and false-positive reactions in the ELISAs were suggested. Thus, it is still advisable to perform both IBT and ELISAs simultaneously in well-defined autoimmune diseases to further analyze the potential advantages of ELISAs using recombinant antigens.

Animals↗
PubMed1987

Site-directed ELISA with synthetic peptides representing the HIV transmembrane glycoprotein.

Two partially overlapping 19 and 22 amino acids long peptides representing a highly immunogenic site of the transmembranous glycoprotein (gp41) of human immunodeficiency virus (HIV) were used as antigen in ELISA tests. The results of antibody determination with this assay were compared with those of three or more conventional ELISAs and Western blot (WB) tests and radioimmunoprecipitation assay. Twenty-six sera from patients with AIDS or LAS and from asymptomatic carriers of HIV infection all showed a pronounced reaction in the peptide ELISA as well as positive results with other tests. In contrast, 27 sera from laboratory workers and blood donors were negative by all tests. A group of 39 blood donor sera, which had shown false positive or ambiguous results in the ELISAs and sometimes in WB tests employed for confirmation, also were negative in all cases with the peptide ELISA. Consecutive samples collected from individuals with primary HIV infection were also analyzed. In 6 out of 9 cases, the peptide ELISA revealed an antibody response within one month after onset of clinical symptoms and sensitivity for antibody detection equaled that of other ELISA tests. Eight sera from five West African persons infected with HIV-related viruses did not react in the peptide ELISA, reflecting differences in properties of the envelope components. The peptide ELISA used in this study appears to represent a simple technique employing chemically synthesized antigen for accurate and sensitive estimation of antibodies to the HIV group of nontransforming human retroviruses.

AIDS-Related Complex↗
PubMedDate not supplied

Diagnostic tests for antiribosomal p protein antibodies: a comparative evaluation of immunoblotting and ELISA assays.

We compared the clinical sensitivity and specificity of three different methods for the detection of serum antiribosomal P protein (anti-P) antibodies in systemic lupus erythematosus (SLE). Sera from 60 unselected SLE patients, 100 healthy subjects and 100 patients with other rheumatic inflammatory diseases were screened for anti-P antibodies by immunoblotting (IB) on P proteins from Raji cells and by two ELISA assays, one using the C-terminal 22 aminoacid long synthetic peptide (C-22) of P proteins, the other using a multiple antigen peptide (MAP) carrying four copies of the C-terminal 13 aminoacid long P peptide.Anti-P antibodies were found in 20% lupus sera by IB, 16.7% by MAP ELISA and 11.7% by C-22 ELISA. The specificity for SLE diagnosis of the three tests in healthy subjects and other rheumatic diseases was: 100% by IB, 100% (vs healthy subjects) and 97% (vs rheumatic diseases) by C-22 ELISA, 100% by MAP ELISA. The agreement between methods was good; differences in concordance rates were restricted to weak positivities. We observed a high concordance in the results of IB and ELISA methods for anti-P antibody detection. IB on P proteins extracted from human lymphoid cells is more sensitive than both ELISAs; IB and MAP ELISA perform better than the C-22 ELISA in determining weakly positive sera.

Adolescent↗
PubMed1994

Use of HPV 1 capsids produced by recombinant vaccinia viruses in an ELISA to detect serum antibodies in people with foot warts.

A sandwich ELISA was developed to detect HPV antibodies using HPV 1 capsids that were purified from recombinant vaccinia virus-infected cells and a monoclonal antibody to the HPV 1 L1 protein. Sera from 91 college-aged women who had been previously screened for HPV 1 antibodies by immune precipitation of capsid proteins were tested by ELISA. A cutoff point was established independently of other criteria based on the assumption that the ELISA values came from a mixture of two Normal distributions representing seropositive and seronegative individuals. It was found that the data fit this model best when the natural log of the ELISA (+0.5 to make all of the values positive) was used. Positive sera were shown to react with a conformational epitope(s) on the L1 protein. In the population reporting foot warts, 16 of 18 (89%) had ELISA values above the cutoff. This compared to 38 of 73 (53%) positives in the population reporting no history of foot warts. The odds ratio for the association of the ELISA reactivity with foot warts was 7.23 (95% CI 1.53, 69.4; P < 0.01). There was no significant association between the ELISA reactivity and wart infections reported at other sites. The average of the log ELISA values for individuals never reporting foot warts was -0.223 (SD 0.468), whereas the average value for individuals reporting foot warts within 10 years was 0.191 (SD 0.450) (P = 0.001). There was a negative correlation between the magnitude of ELISA reactivity and the time elapsed since the last appearance of foot warts. This apparent loss of seroreactivity over time may indicate that HPV 1 is usually eliminated from the host after infection or that inadequate levels of HPV 1 capsid antigen are produced during latent foot warts to maintain antibody levels.

Adolescent↗
PubMed1992

Improved detection of antibodies to hepatitis C virus using a second generation ELISA.

A screening assay for the detection of antibodies to hepatitis C virus (HCV); ORTHO HCV ELISA Test System, Second Generation, was compared with the currently licensed c100-3 based test (ORTHO HCV ELISA Test System). The second generation ELISA differs from the c100-3 based assay in that it detects circulating antibodies to both structural (nucleocapsid) and non-structural (NS3/NS4) HCV proteins. Specimens tested consisted of a cohort of 35 patients diagnosed with non-A, non-B hepatitis (NANBH) and 3971 presumably healthy volunteer blood donors. Second generation ELISA demonstrated significantly greater clinical sensitivity in patients with acute phase NANBH (80% vs. 60%) as well as chronic disease (88% vs. 72%). Additional specimens reactive only in second generation ELISA, demonstrated reactivity to HCV antigens c33c and/or c22-3 in supplemental testing by the Chiron HCV RIBA Assay System. The second generation ELISA also detected additional RIBA reactive volunteer blood donors (0.18% of the population tested) that were nonreactive in first generation ELISA. This data indicated that second generation ELISA would detect approximately 2 additional anti-HCV reactive donors per 1,000 screened. Specificities obtained with this low risk population were 99.6% for first generation and 99.7% for second generation ELISA.

Blood Donors↗
PubMed1995

Pristane induces high titers of anti-Su and anti-nRNP/Sm autoantibodies in BALB/c mice. Quantitation by antigen capture ELISAs based on monospecific human autoimmune sera.

Autoantibodies to Su and anti-nRNP/Sm are common in human and murine systemic lupus erythematosus (SLE), and are also produced by BALB/c mice with SLE-like autoimmunity induced by pristane. Antigen capture ELISAs employing monospecific human autoimmune IgG were developed to quantitate the production of anti-Su and anti-nRNP/Sm autoantibodies in 77 sera from BALB/c mice with pristane-induced autoimmunity. The sensitivity and specificity of the anti-Su antigen capture ELISA were 100% compared with immunoprecipitation of 35S-labeled cellular proteins. All 16 immunoprecipitation positive sera were positive in the anti-nRNP/Sm antigen capture ELISA (100% sensitivity), whereas 55/61 immunoprecipitation negative sera were negative by ELISA (90% specificity). The 6/61 immunoprecipitation negative sera that were ELISA positive were probably true positives because subsequent sera obtained from the same mice were positive by both techniques. Thus, the antigen capture ELISA may be somewhat more sensitive than immunoprecipitation. The titers of anti-Su and anti-nRNP/Sm positive antibodies in the sera were as high as 1:25,000-1:250,000 by ELISA, suggesting that autoantibodies may be produced in pristane-primed BALB/c mice at levels comparable to those seen in spontaneous autoimmune disease. We conclude that antigen capture ELISAs based on human autoimmune sera were highly sensitive and specific for detecting murine anti-Su and anti-nRNP/Sm antibodies. This technique will be useful for quantitating antibodies in murine autoimmune disease models, since antigen capture ELISA avoids the use of denatured or recombinant antigens, permitting antibodies recognizing tertiary and quaternary structures to be detected.

Animals↗
PubMed1988

Enzyme-linked immunosorbant assay (ELISA) of size-selected crotalid venom antigens by Wyeth's polyvalent antivenom.

The binding of Antivenom (Crotalidae) Polyvalent to fractions from crude venoms of eight crotalid and one viperid snake, obtained by high performance size-exclusion chromatography, was determined with an indirect enzyme-linked immunosorbent assay (ELISA). Most of the large (greater than 30,000 mol. wt) molecular mass crotalid venom fractions were associated with high (greater than 0.7 absorbance units) ELISA values. Similarly, the medium (13,000-30,000 mol. wt) and small (less than 14,000 mol. wt) molecular mass crotalid venom fractions were coincident with moderate (0.3-0.7 absorbance units) and low (less than 0.3 absorbance units) ELISA levels. Some variability in this pattern was seen with individual venom fractions. A distinctly different pattern of ELISA values were observed with two rattlesnake venoms: the South American (Crotalus durissus terrificus) and Mojave desert (Crotalus scutulatus scutulatus) rattlesnakes. The elution profile from these venoms showed a progression of low to moderate ELISA values within the large molecular mass fractions. This pattern was followed by a decline to low ELISA values throughout the remainder of the elution profile. When saw scaled viper (Echis carinatus leucogaster) venom fractions were tested, only background ELISA values were detected with antivenom. Similarly, background ELISA values were associated with the small molecular mass fractions of all venoms tested. In addition, the elution position for the basic peptides of southern Pacific (Crotalus viridis helleri) and timber (Crotalus h. horridus) rattlesnake venoms showed minimal ELISA values. These data support the view that except for the venom of C. durissus terrificus and C. s. scutulatus, most antivenom antibodies bind large (greater than 30,000 mol. wt) venom fractions. Thus, antivenom contains minimal levels of antibodies to the basic peptides in these venoms.

Animals↗
PubMed1987

Modification of the ELISA for the estimation of tetanus antitoxin in human sera.

The use of indirect ELISA for the quantitation of tetanus toxin neutralizing antibodies in human sera is limited by marked overestimations in low titered sera. The reasons for the discrepancy between the results obtained by ELISA and by in vivo assay and modifications of the ELISA to overcome the problem were investigated. Catching ELISA and indirect ELISA using trays coated with the contaminant proteins in toxoid preparations indicated that antibodies to contaminants were only partly responsible for the discrepancy and the introduction of these modifications did not solve the problem. In ELISA competition experiments with toxin neutralizing monoclonal antibodies, the human immunoglobulins irrelevant in toxin neutralization, but detectable in indirect ELISA, were found to be difficult to inhibit in their binding to the solid antigen phase. These might represent antitoxins bound bivalently to the solid phase but with affinities in monovalent binding insufficient for toxin neutralization or other coupled antibodies due to conformational changes of the antigen. A competition ELISA with toxin in solution was therefore developed to assess selectively the antitoxin capable of binding the antigen in solution and by this approach the in vivo activities of even low titered sera were accurately predicted. This antigen competition ELISA may be easily introduced into routine tetanus serology and the principle may also be of value for the in vitro detection of functional antibodies to other antigens.

Antibodies, Monoclonal↗
PubMed1996

Ovine lentivirus antibody detection in serum, colostrum and milk using a recombinant transmembrane protein ELISA.

An enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies against ovine lentivirus (OLV) in serum, colostrum, and milk from naturally infected sheep. The assay used OLV recombinant transmembrane envelope protein (rTM) as a test antigen. Matched serum/colostrum and serum/milk samples were collected at 24h, 4 weeks (mid-lactation), and 8 weeks (weaning) post-lambing. Among 129 paired samples collected at 24 h post-lambing, there was overall test agreement (concordance) of 82.9% and a kappa value of 0.658 between serum and colostrum rTM ELISA results. Among 130 mid-lactation samples, the milk ELISA had 100% specificity and 64.9% sensitivity relative to the serum ELISA, there was concordance of 79.2%, and a kappa value of 0.602. At mid-lactation, the serum agar gel immunodiffusion test had a sensitivity of 0.390 and 0.560 relative to the serum and milk rTM ELISAs, respectively. Matched serum and milk rTM ELISA results at weaning were very similar to those at mid-lactation. Finally, increased occurrence and severity of subclinical mastitis at weaning was found in ELISA-seropositive compared with ELISA-seronegative ewes. Both subclinical mastitis and ewe OLV infection had a negative impact on lamb growth and weaning weights. Compared with blood, colostrum and milk are easier and less expensive to sample and store. These results suggest that rTM ELISA testing of colostrum and milk could be used to supplement serologic testing in OLV screening or eradication programs.

Animals↗
PubMed1985

Plant virus detection using a new form of indirect ELISA.

A novel form of indirect enzyme-linked immunosorbent assay (ELISA) has been devised for the detection of viruses in plants. The method uses protein A in two applications to sandwich antibody-antigen-antibody layers. The first applied layer of protein A prepares the plate for the coating antibody layer. The second layer of protein A is conjugated to the enzyme and detects the second antibody layer. The orientation of the IgG induced in the coating layer of antibody prevents later unwanted reaction with the conjugated protein A. Using seven antisera, protein A sandwich ELISA (PAS-ELISA) detected homologous virus isolates in standard dilutions of infected plant homogenates at A405 values which were at least one absorbance unit greater than those of healthy controls. The PAS-ELISA method was more sensitive than the direct double antibody sandwich form of ELISA (DAS-ELISA), e.g. not only were A405 values for homologous reactions greater in PAS-ELISA but also an antiserum to a birch isolate of cherry leaf roll virus detected four related isolates with the new method against only one with DAS-ELISA. However, dilution end points for the homologous virus were about the same in both methods. In a practical application, PAS-ELISA detected prune dwarf virus in 18-36% of tested Prunus avium seeds.

Animals↗
PubMed1994

Suitability of autoclaved tap water for preparation of ELISA reagents and washing buffer.

The suitability of autoclaved tap water for the preparation of ELISA reagents and washing buffer was compared with that of ultrapure water, in a standard indirect ELISA for the detection of antibodies to pseudorabies virus (PRV). The performance of the assay, using autoclaved tap water (AT-ELISA) compared favourably to that of the standard assay, using ultrapure water (UP-ELISA) in detecting anti-PRV antibodies in sequential serum samples from a pig experimentally infected with PRV. While both the UP-ELISA and AT-ELISA proved reliable in detecting anti-PRV antibodies in a coded proficiency serum panel (n = 60), the AT-ELISA detected fewer positive sera than the UP-ELISA in evaluating a limited number (n = 80) of field samples. The results suggest that autoclaved tap water may be substituted for ultrapure water for the preparation of ELISA reagents when or where ultrapure water may not be available.

Animals↗
PubMed1988

Preparation and evaluation of the specificity of Parafilaria bovicola antigen for detection of specific antibodies by ELISA.

An enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies in bovine sera against Parafilaria bovicola nematodes was developed and its sensitivity was compared with the immunodiffusion (ID) method. An exoantigen of P. bovicola which was shown to contain four major polypeptides was used in these procedures. The serological reactivity of the antigen polypeptides was defined by using the enzyme-linked immunoelectrotransfer blot technique (EITB) and whole-worm extract proteins. It identified only four serologically reactive polypeptides with sera from one experimentally infected calf and a verified field case. These two positive sera reacted mainly with four major antigens which coincided in molecular weights of the polypeptides of the exoantigenic preparation, namely, 43, 39, 28 and 25 KDa. Calves experimentally infected with P. bovicola showed a positive reaction with ELISA at 4 months after inoculation, and after this period a rapid increase in serum antibody response occurred. In these cases the ID reaction was observed for the first time at 7 months after inoculation. The specificity of an ELISA method using crude exoantigen preparation of P. bovicola was tested for the diagnosis of bovine parafilariasis. No cross-reactivity was detected when the P. bovicola exoantigen preparation was tested against sera from calves experimentally infected with Onchocerca lienalis, as well as against the sera from cattle naturally infected with Dictyocaulus viviparus or from cattle chronically infected with Ostertagia ostertagi. In addition, testing of 740 field sera from cattle in areas non-endemic and endemic for P. bovicola indicated a specificity of the antigen preparation used. Forty sera from laboratory-confirmed field cases of P. bovicola infection were tested by ELISA and immunodiffusion. All of these sera were ELISA positive, whereas only 70% of these were positive in the ID test. Seven (2.1%) of 328 sera from 21 herds from non-endemic P. bovicola areas were ELISA positive, as opposed to none in the ID test. Of the 94 sera from six herds in areas endemic for P. bovicola infection, 51 (54%) were ELISA positive whereas only 24 (26%) were positive in the ID test. When 56 slaughtered cattle, with varying degrees of meat condemnations due to parafilariasis, were tested for P. bovicola specific antibody, 91% of the serum samples were positive by ELISA. These results suggest that the exoantigen of P. bovicola can be used in a sensitive and reliable serological detection of parafilariasis by ELISA.

Animals↗
PubMedDate not supplied

ELISA for detection of IgM and IgG antibodies to sandfly fever Sicilian virus.

An enzyme-linked immunosorbent assay (ELISA) was developed to detect specific human immunoglobulin G and M antibodies to sandfly fever Sicilian (SFS) virus. Acute and early convalescent serum pairs with greater than or equal to 7 days between the 2 specimens were available from 20 patients and all showed significant optical density (OD) increase and significant titre rise (greater than or equal to 4-fold) by IgG ELISA. However, negative or borderline-positive sera were found as late as 11 days after onset of symptoms when tested by IgG ELISA. Specific IgM antibodies were detected during the first week of symptoms, and maximum OD values were obtained during the first 4 weeks after onset of disease. The IgM OD values declined over the following 3-9 months. All sera collected later than 14 months post-onset were negative by IgM ELISA. The combination of early antibody response and the need to test only one serum specimen gives IgM ELISA an advantage over IgG ELISA in patients diagnosis. The IgG ELISA was also evaluated as a seroepidemiological tool and compared to a plaque reduction neutralization test (PRNT) using sera from a normal Cypriot population. Of 183 sera tested, 34 (19%) were positive in plaque reduction neutralization tests (PRNT) and 113 (62%) by IgG ELISA. A number of PRNT-negative sera were strongly positive by IgG ELISA and also by indirect immunofluorescence test, which may suggest the presence of a virus related to SFS in Cyprus which has not yet been isolated.

Acute Disease↗