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PubMed2003

Validation of FPA and cELISA for the detection of antibodies to Brucella abortus in cattle sera and comparison to SAT, CFT, and iELISA.

The fluorescence polarisation assay (FPA) is a recently described test for the serological diagnosis of Brucella infection. It has many methodological advantages over older, more established tests and can be performed in a fraction of the time. To validate the FPA, serum samples from 146 confirmed (by culture) Brucella-infected cattle were tested in conjunction with serum samples from 1947 noninfected cattle. The competitive ELISA (cELISA) was validated using these positive reference samples and 1440 negative samples, while data for the indirect ELISA (iELISA) was generated from 6957 negative samples plus the positive sera. Published diagnostic specificity (DSp) data for the complement fixation test (CFT) and serum agglutination test (SAT) was used in conjunction with the test results on the positive sera to obtain diagnostic specificity plus diagnostic sensitivity (DSn). After selection of a cutoff for the FPA and cELISA, the diagnostic specificity and sensitivity total for each test were compared. The results, with 95% confidence intervals, were: FPA (195.7+/-2.79), iELISA (195.0+/-2.70), cELISA (194.9+/-3.48), CFT (191.7+/-4.45), and SAT (180.4+/-6.33). The data presented supports the use of the FPA in diagnosis of brucellosis and questions the use of the SAT and CFT for either screening or confirmatory testing.

Agglutination Tests↗
PubMed1979

Solid-phase radioimmunoassay techniques for the detection of African swine fever antigen and antibody.

A solid phase radioimmunoassay (RIA) has been successfully developed to measure both African swine fever virus (ASFV) antigen and antibody. Studies show that the assay is reproducible and will detect limiting antigen concentrations equivalent to 50--500 HAD50/ml. Both direct and indirect antibody RIA have been developed and have proved to be approximately 100 times fore sensitive than the complement fixation test at present available and 1000 times more sensitive than the immuno-electro-osmophoresis test for the detection of ASFV antibody.

African Swine Fever Virus↗
PubMed1995

[Evaluation of conventional and new generation tests for testing the humeral response to Mycoplasma pneumoniae antigens in natural infection in humans. I. Occurrence and level of mycoplasma antibodies in clinically healthy subjects].

The aim of the first stage of investigation was to determine, using the complement fixation test (CFT), immuno-electroprecipitation test (IEPT) and ELISA, the level of mycoplasmal antibodies in serum samples obtained from 114 clinically healthy persons (49 samples from children and youth up to 20 and 65 samples from blood donors). On the basis of obtained findings, antibody titer assumed to be diagnostically significant was determined on the means + 3 SD level in each test. Thus in the CF test the assumed titer > or = 20, in the IEP test--titer > or = 2, in the ELISA-IgA--titer > or = 100, in the ELISA- IgG--titer > or = 1600, in the ELISA-IgM--titer > or = 1200, in the ELISA -IgA + G + M--titer > or = 3200. The study allowed to establish that on with age the level of mycoplasmal antibodies in immunoglobulin of classes A and G rose while the titer of these antibodies of class IgM remained on the same level in persons up to 20 and then decreased almost twice in adults. It was also found that in all age groups the mean values of titer of antibodies against M. pneumoniae calculated for each test were lower than titer assumed to be diagnostically significant.

Adolescent↗
PubMed1991

Comparative evaluation of enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of dourine.

The detection of antibodies against Trypanosoma equiperdum in 689 equid sera was compared by enzyme-linked immunosorbent assay (ELISA), the complement fixation test (CFT) and an indirect immunofluorescent test (IIF). CFT was the least sensitive technique, susceptible to anti-complementary factors and the most technically demanding. IIF was more sensitive, but was only suitable for testing limited numbers of samples. In this study, ELISA was the most sensitive test, the least labour intensive and lends itself to a considerable degree of automation. It is suggested that ELISA would be relatively easy to standardise between laboratories and an ELISA protocol could be adopted as the internationally approved test for equine health certification.

Animals↗
PubMedDate not supplied

Evaluation of eight antibody tests and one antigen test for the diagnosis of invasive aspergillosis.

Eight Aspergillus antibody detection assays--three indirect haemagglutination assays (IHA-LD, IHA-Roche, IHA-Fumouze), three enzyme immunoassays (EIA-IgG, EIA-IgM, EIA-IgA, DDV) and two complement fixation tests (CF-metabolic and CF-somatic, Virion--and one latex agglutination test (LAT) for Aspergillus galactomannan antigen detection (Sanofi Pasteur) were evaluated in 14 patients with proven invasive aspergillosis (a total of 47 serum samples and one cerebrospinal fluid sample) and in 68 selected control individuals (one selected serum sample each). For the antibody tests, sensitivity ranged from 14% to 36% and specificity from 72% to 99%. The antigen detection test had a sensitivity of 36% and a specificity of 100%. Currently commercially available antibody detection assays for the serodiagnosis of invasive aspergillosis are inadequate. The antigen detection test appears to be highly specific, but lacks sufficient sensitivity.

Adult↗
PubMed1981

Experimental bovine trypanosomiasis (Trypanosoma vivax and T. congolense). III. Serum levels of immunoglobulins, heterophile antibodies, and antibodies to T. vivax.

Twenty-five steers were infected with T. vivax (EATRO 1721), 25 steers with T. congolense (EATRO 1800) and 25 steers kept as controls. Serum levels of immunoglobulins (IgG1, IgG2, IgM), natural antibodies to erythrocytes of chicken and sheep, and complement-fixing antibodies to T. vivax were measured. A significant decrease of serum IgM and natural antibodies to chicken erythrocytes occurred in the T. vivax group at day 9, i.e. at the decline of the first parasitemic wave. This was followed by a transient moderate increase of IgM accompanied by a transient decrease of IgG2. The T. congolense group had moderate decreases (less than 30%) of the mean IgG1 levels and moderate increases (less than or equal to 50%) of the mean IgG2, levels. It was concluded that there was little evidence for polyclonal activation of lymphocytes and that the decreased IgG1 levels in the T. congolense group might have been a reflection of immunosuppression. The complement-fixation test proved to be a sensitive tool for monitoring the antibody response to T. vivax.

Animals↗
PubMed1976

The diagnostic value of determination of IgM antibodies against Mycoplasma pneumoniae by the indirect immunofluorescent antibody test.

The indirect fluorescent antibody technique for specific IgM and IgG antibodies was applied to paired sera from 33 patients with a current M. pneumoniae infection, along with the complement fixation test. A roughly parallel increase in antibody titres in all the three tests was observed; the ratio of IgM/IgG titres was, however, higher in patients below 20 years of age than in older patients. Rises in antibody titres were regularly observed in spite of the fact that most of the patients were treated with tetracycline or erythromycin during the acute phase of the disease. Serum specimens from 15 other patients lacking clinical data of a recent M. pneumoniae infection, but with stationary complement fixation titres, all showed IgG antibodies with stationary titers and, with the exception of three patients, a negative IgM antibody test. Sera from 20 individuals lacking complement fixing antibodies were also without demonstrable IgG and IgM antibodies. Sampling of serum from 9 of the pneumonia patients was repeated. The last IgM positive sample was collected up to six months after onset of the disease and all were found to be negative in the IgM test 8 to 10 months after onset of the illness in these treated patients. The implications of these findings for the serological diagnosis of M. pneumoniae infections are discussed.

Adolescent↗
PubMed1973

Some observations on the serology of Pneumocystis carinii infections in the United States.

An indirect immunofluorescent (IF) test with antigen of partially cleaned Pneumocystis carinii cysts was evaluated for sensitivity and specificity. The test was reactive at the 1:8 and above level for 44% of the persons who had suspected and confirmed infections, for 7.3% of the healthy contacts, and for 1.4% of the general population. Results of tests with sera from people with measurable antibodies to other diseases and those on immunosuppressant therapy suggest that only titers of 1:20 and above are specific. At this level, tests with sera of 32% of the cases, 4% of the contacts, 0% of the healthy controls, and from 0% to 37% (average 5%) of individuals on long-term immunosuppressive therapy or with antibodies to other diseases were positive. The high percentage of positive reactions in cytomegalovirus and in fungal infections could represent double infections. The complement fixation test was less sensitive than the IF test. Antibodies measured by the IF test were immunoglobulin G. No differences in antigenicity were demonstrated by the IF test among the cysts from eight human infections or from induced rat infections.

Animals↗
PubMed2002

Humoral immune responses following experimental infection of goats with Mycoplasma capricolum subsp. capripneumoniae.

Goats housed in microbiologically secure facilities were experimentally endobronchially infected with Mycoplasma capricolum subsp. capripneumoniae (Mccp), causal agent of contagious caprine pleuropneumonia (CCPP). The animals were monitored over an 8-week period post-infection (p.i.). Elevated temperatures were observed 2-7 days p.i., reaching a maximum of 41.5 degrees C in one animal (1884). By 8 weeks p.i. the infection was successfully cleared, with no Mccp being recovered from the lungs, serum or nasal passages. Mccp was not isolated from serum throughout the experiment, either directly by culture or indirectly via polymerase chain reaction (PCR). Humoral immune responses against Mccp capsular polysaccharide (CPS) were generally poor when measured by ELISA. CPS antigen was present in the serum of all infected animals early in the infection (day 14 p.i.), although in one animal (1855) CPS antigen persisted throughout. This was the only animal to exhibit a serious cough (day 5-19 p.i.). Successful diagnosis of CCPP was achieved using two different types of latex agglutination test (CPS antibody and CPS antigen detection test), immunoblotting and a blocking ELISA, although the latter lacked sensitivity until later in the infection (35-40 days p.i.). Only a single animal (1855) was detected positive using the current complement fixation test (CFT). Strong immune responses to protein antigens were detected by IgG and IgM immunoblotting from the first time point at day 14 p.i. IgM immunodominant bands of 220, 85, 62 and 40kDa were observed in the 3 infected animals and from CFT-positive CCPP field sera. Band intensity gradually diminished throughout the experiment. IgG immunodominant bands of 108, 70, 62, 44, 40 and 23kDa were shared between experimentally-infected and field sera, with band intensity either remaining unchanged or increasing from day 14 p.i. These bands were not present using pre-infection sera. Of the diagnostic tests used, only the CPS antibody detection latex agglutination test and IgG immunoblotting gave positive diagnoses throughout the entire period post-infection (days 14-53 p.i.).

Animals↗
PubMed1974

Production and reactivity of immune sera specific for HADEN virus polypeptide antigens.

Antisera were prepared against the three structural polypeptides of HADEN virus dissociated with sodium dodecyl sulfate. These immunological reagents were used in immunofluorescence tests to study the kinetics and location of polypeptide antigen appearance in infected cells. These sera did not neutralize virus infectivity, did not cross-react with adenovirus-associated virus-infected cells, and reacted in complement fixation tests with sodium dodecyl sulfate-dissociated virus, but not with complete virus antigen. The polypeptide antigens were heat labile, and all appeared in infected cells at least 2 h prior to whole-virus antigen.

Adenoviridae↗
PubMed1980

Differentiation of porcine enterovirus serotypes by complement fixation.

A complement fixation test in microtitre plates for the differentiation of porcine enterovirus serotypes is described, employing guinea pig antisera prepared using inactivated purified viruses. Eleven porcine enterovirus serotypes and swine vesicular disease virus were compared and clearly distinguished from each other. In addition, 71 porcine enterovirus strains and isolates were tested and each was identified as belonging to one of the 11 serotypes.

Animals↗
PubMed1992

Productivity and health of camels (Camelus dromedarius) in Somalia: associations with trypanosomosis and brucellosis.

In Somalia, one of the world's largest dromedary populations of about 5.3 million animals are kept by nomadic pastoralists under traditional management. Interest in the development potential of camel herds in the semi-arid areas of central Somalia initiated an investigation to determine the productivity of herds, their major diseases and likely associations among these parameters. Using a systems approach, data were collected for herd production parameters, environmental factors, management and production systems, and health variables. One thousand and thirty nine camels in 33 herds were studied in the central regions of Somalia. Trypanosoma evansi prevalence ranged from 1.7% in blood-smears to 56.4% using enzyme-linked immunosorbent micro-assay (microELISA). Seroprevalence for brucellosis was determined as 1.9% by the standard agglutination test (SAT) and 0.3% by the complement fixation test (CFT). Using multiple regression, 15% of the total variation of the general fertility rate was explained by the results of the microhaematocrit centrifugation technique (MHCT) and the microELISA for T. evansi, CFT results for brucellosis, herdsize, and young stock death rate. Among herd production variables, herd size differed significantly for different management units. Young stock death rates, as well as general fertility rates varied in the ecological subzones with a marked effect in the zones labeled "Inland". Various other associations were noted among demographic, husbandry and disease variables. The importance of trypanosomosis and brucellosis to the productivity of herds and measures to control their limiting effects on production were discussed.

Animal Husbandry↗
PubMed1964

MEASLES MYOCARDITIS.

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Alanine Transaminase↗
PubMed1979

The bovine immune response to Brucella abortus. II. Elimination of some sporadic serological reactions by chelation of divalent cations.

The standard agglutination tests for detecting antibody to Brucella abortus were modified by addition of chelating agents (EDTA and EGTA) to the antigens. Approximately 80% of "singleton" agglutination test reactions, negative on the diagnostic complement fixation test, obtained with cattle sera were eliminated while no decrease in titer was apparent when sera from B. abortus infected or vaccinated cattle were tested.

Agglutination Tests↗
PubMed1956

Use of synthetic, crystalline, L-alpha-dimyristoyl lecithin in cardiolipin antigens.

Experiments were carried out by the authors to determine whether synthetic, crystalline, L-alpha-dimyristoyl lecithin could replace natural purified lecithins in the preparation of cardiolipin antigens. These experiments were designed specifically to find out whether it was possible to obtain the same serological reactions, qualitatively and quantitatively, with the test antigen as with a reference antigen containing natural lecithin, and whether the test antigen had the same keeping qualities as the reference antigen.The tests used were the quantitative complement-fixation test as modified by Mørch in 1933, and the VDRL slide flocculation test.The results showed that synthetic, crystalline, L-alpha-dimyristoyl lecithin could replace natural lecithin in the preparation of cardiolipin antigens, but that the antigens prepared with the synthetic lecithin were significantly less sensitive than those prepared with an equimolar amount of natural lecithin. The authors consider that further investigation is required before the use of synthetic lecithin is finally adopted.

Cardiolipins↗

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