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Haemophilus somnus: a comparison among three serological tests and a serological survey in beef and dairy cattle.

Serological tests for the detection of antibodies against Haemophilus somnus were carried out in herds of beef and dairy cattle using three different techniques: agglutination, complement fixation and counterimmunoelectrophoresis. The agglutination test appeared to detect more seroreactors than the complement fixation and counterimmunoelectrophoresis tests. Results of the three tests indicated that there were more positive reactors in beef cattle and dairy cattle from infected herds than in dairy cattle from clinically normal herds.

Agglutination Tests↗

Electroaddressed immobilization of recombinant HIV-1 P24 capsid protein onto screen-printed arrays for serological testing.

A serological chemiluminescent biochip was designed based on screen-printed electrode arrays composed of nine 1-mm(2) electrodes. Arrays were shown to be produced with good batch-to-batch reproducibility (standard deviations of 4.4 and 12.0% for ferricyanide oxidation potential and current, respectively) and very good reproducibility within a particular array (2.0 and 7.5% standard deviations for the same controls). Electrode arrays were used to electroaddress various bioconjugate structures comprising a recombinant HIV-1 P24 capsid protein (RH24K) in polypyrrole film. Entrapment of RH24K preimmobilized onto maleic anhydride-alt-methyl vinyl ether copolymer was shown to be the more efficient immobilization procedure. This addressed sensing layer enabled the detection of anti-P24 antibodies at a concentration of 3.5 ng/ml through peroxidase-labeled anti-human immunoglobulin G reaction. The biochip was used to perform an HIV-1 serological test in human sera. HIV-1 seropositive and seronegative sera were easily discriminated using serum dilutions greater than 1/10,000.

Amino Acid Sequence↗

[Urease test, microbiologic tests, histologic and serologic tests for the evaluation of Helicobacter pylori infections in persons with peptic ulcer and gastritis].

The presence of Helicobacter pylori in the gastric mucosa of 150 patients with gastric, duodenal ulcer and endoscopic gastritis was investigated by rapid urease-based test, culture, histological examination and serology. Overall, 78% were positive for Helicobacter pylori by rapid urease-based test, 69.3% by culture, 53.7% by histological staining and 93.2% by serology (p < 0.001). The frequency of Helicobacter pylori was higher in patients with gastric or duodenal ulcer compared with those with endoscopic gastritis and those after gastric resection. However, the differences were significant only with reference to rapid urease-based test, (p < 0.02). The EIA (Roche) serological test had the highest sensitivity and the lowest specificity. This indicates the seed for serological kit to be independently evaluated on the population to be studied. The rapid urease-based test was simple, rapid and inexpensive, and it was more sensitive and equally specific compared with culture and Giemsa stain.

Adolescent↗

The prevalence of antibodies against B. burgdorferi, an indicator for Lyme borreliosis in dogs? A comparison of serological tests.

Five serological tests for the detection of IgM and IgG antibodies to Borrelia burgdorferi, the causative microorganism of Lyme borreliosis (LB), were compared in 1177 sera from Dutch dogs: 401 healthy working hunting dogs, 100 healthy city pet dogs, 629 city dogs suspected of having LB with various clinical symptoms, and 47 hunting dogs with intermittent lameness. The results of the in-house species-independent enzyme immunoassay (i.e. an EIA which can be used to test serum samples from different animal species) showed a strong agreement (kappa: 0.78-0.81) with those of an experimental and a commercially available EIA (Genzyme Virotech, Rüsselsheim, Germany) for the detection of canine IgG antibodies to B. burgdorferi. Furthermore, the sensitivity of the in-house EIAs for the detection of antibodies to B. burgdorferi was independent of the antigenic heterogeneity, as demonstrated by the results of sera from dogs suspected of LB with various clinical symptoms: lameness (n = 60), and neurological (n = 60) and skin disorders (n = 52). Because of its high sensitivity for IgM antibodies, the indirect assay (Diagast, Pessac, France) proved to be an interesting tool for the detection of an acute Lyme infection in dogs. However, in this study a positive serological result could not be linked to any clinical symptom that has been related to LB in dogs. Results showed no difference in seroprevalence between dogs considered at high or at low risk of a B. burgdorferi infection. It was concluded that LB is an uncommon disease in the Dutch dog population despite the fact that many of Dutch dogs are infected with B. burgdorferi. Because of this low prevalence, the use of any immunoassay to support the clinical diagnosis of LB in dogs might be of limited value. Nevertheless, the species-independent EIA could be valuable in seroepidemiological studies when sera of several different animal species need to be tested.

Animals↗

Antibody dynamics in Holstein Friesian heifers vaccinated with Brucella abortus strain 19, using seven serological tests.

The serological response induced by Brucella abortus strain 19 was evaluated in 52 Holstein females from a brucellosis-free herd using seven serological tests. Each calf was vaccinated at an age of 4 and 8 months old with 3 x 10(10) CFU B. abortus S19 and the antibody response was determined as the proportion of positive results to each test. The antibody dynamics, measured with the buffered plate antigen (BPA) test and the rapid automated presumptive (RAP) test, were similar. The proportion of positive reactions in these tests reached 100% one week after vaccination and remained at this level for seven weeks, after which the proportion of positive samples slowly declined to 8% (BPA) and 2% (RAP) at week 50. The response in the indirect enzyme immunoassay (i-ELISA) was similar, but shorter than that observed with the BPA/RAP. The antibody dynamic, measured using the seroagglutination test (SAT) in parallel with the 2-mercaptoethanol (2-Me) test and the complement fixation test (CFT) were similar to the RAP/BPA, but of slightly shorter duration. The competitive ELISA (c-ELISA) was positive in all animals for 3 weeks, followed by a rapid decline. The fluorescence polarization assay (FPA) reached a maximum of 68.5% positive animals at week 4 and then declined. Based on these data, the c-ELISA and FPA discriminated residual antibody activity due to vaccination more efficiently than the other tests.

Animals↗

Comparison of a latex agglutination test with other serological tests for the measurement of antibodies to Toxoplasma gondii.

One hundred sera from 49 patients with glandular toxoplasmosis were examined by a latex agglutination test, the dye test, an indirect haemagglutination test, and a double antibody sandwich enzyme linked immunosorbent assay (ELISA) for antitoxoplasma IgM. The results support previous findings that the dye test, indirect haemagglutination test, and latex agglutination test measure different antibodies to Toxoplasma gondii. In early glandular toxoplasmosis, when specific IgM was detected, the titres of both the latex agglutination test and the indirect haemagglutination test were lower than the dye test. Repeat specimens from 11 of the patients showed four cases in which the latex agglutination test titres never exceeded 1/256, whereas both the dye test and the indirect haemagglutination test showed significant titres and specific IgM was detected in every case. We conclude that the latex agglutination test should not be used as a substitute for the dye test in the serological diagnosis of glandular toxoplasmosis. All sera giving a positive latex agglutination test result should be referred for further tests. A combination of the dye test and double antibody sandwich ELISA gives the most reliable serological diagnosis of early glandular toxoplasmosis.

Antibodies↗

Development, implementation, and impact of acceptability criteria for serologic tests for infectious diseases.

Serologic testing is essential for the diagnosis of some infectious diseases and yet is fraught with potential pitfalls. All parts of the diagnostic process must be optimized to ensure that serologic tests perform adequately. Recognizing that a lack of clinical data and correctly timed, paired sera frequently led to uninterpretable serology results at our laboratory, we developed and implemented simple acceptability criteria for serologic tests. We assessed the impact of these criteria by comparing submissions and results for the year before and the year after implementation of the criteria. The number of serologic tests performed declined by 25% after implementation of the acceptability criteria, despite an increase in requests for serologic tests. Inappropriate testing of acute-phase sera alone fell from 49 to 0% (P < 0.001) for the tests monitored. Appropriate submission of paired sera rose from 9 to 19% (P = 0.006). The proportion of results classified as interpretable rose from 52 to 100% (P < 0.001). We recommend that acceptability criteria be developed and applied to samples submitted to clinical microbiology laboratories for serologic testing.

Bacterial Infections↗

Agar-gel precipitin-inhibition test for coccidioidomycosis. II. Serological test antigen studies in agar-gel.

In this study of saprophytic and parasitic growth-phase extracts of Coccidioides immitis, an antigen from the spherule culture supernatant fluid detected a specific antibody in heretofore serologically negative suspect coccidioidomycosis human sera when diffused in agar-gel. This antigen-antibody reaction occurred also in some of the serologically positive human coccidioidomycosis sera. This study indicates that this antigen-antibody reaction should be utilized as a possible routine serological test for complete serodiagnosis.

Antibodies↗

The serological responses of calves to live Salmonella dublin vaccine--a comparison of different serological tests.

The serological responses to live Salmonella dublin vaccine was assessed in three groups of calves; three-day-old colostrum-deprived (3DO C-), three-day-old colostrum-fed (3DO C+) and three-month-old (3MO), by the following tests; serum agglutination test (SAT), indirect haemagglutination test (IHA), complement-fixation test (CFT) and antiglobulin test (AGT). Serological activity was detected by all the tests in the 3MO calves. In the 3DO C+ calves no serological activity was detected by either the somatic SAT or IHA but low levels of CF and somatic AGT antibodies were produced. In 3DO C- calves serological activity, often at low levels, was detected by all the tests except the somatic SAT. High levels of flagellar agglutinins were detected in both groups of 3DO calves. It was concluded that with the exception of the flagellar SAT the tests were affected by the age of the calf and in 3DO calves also by the presence of colostral antibodies. However, the use of the SAT in 3MO calves would provide an indication as to the potency of salmonella vaccines.

Agglutination Tests↗