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1.
A comparison was made between serological and parasitological techniques for the diagnosis of bovine trypanosomiasis in Zambia. Overall sero-prevalence rates as determined by IFAT and ELISA were respectively 2.7-fold and 2.9-fold greater then the percentage of samples found positive with the dark ground/phase contrast buffy coat technique (DG). The results obtained by the two serological techniques were found to be closely correlated (94.2%) agreement) and titres obtained by ELISA tended to be slightly higher than those obtained by IFAT. Linear regression analysis of the results obtained by the IFAT and DG techniques revealed a highly significant correlation. This finding would permit the use of only one of the techniques in an epidemiological survey and to extrapolate the results from the regression line.  相似文献   

2.
This work reports the results obtained by a new enzyme-linked immunosorbent assay (ELISA) test developed for the serological diagnosis of canine leishmaniasis.The new ELISA is based on a recombinant protein obtained by joining different antigens of Leishmania infantum.Test performances have been evaluated through the screening 227 sera of dogs, infected and uninfected by L. infantum. The new ELISA test has been compared to the indirect immunofluorescent-antibody test (IFAT) as a reference assay of canine leishmaniasis, and to a commercial ELISA.Excluding from the total number of IFAT positive sera the 27 sera with IFAT titre 1:40 (considered doubtful), the recombinant ELISA showed 97.0% specificity, 93.9% sensitivity and 95.5% agreement with IFAT. The commercial ELISA showed 78.2% specificity, 94.9% sensitivity and 86.5% agreement with IFAT.The results demonstrate a higher performance of the new recombinant ELISA test for the detection of negative samples, with a greater agreement with the reference test (IFAT).  相似文献   

3.
A total of 173 aborted ovine foetuses and seven aborted caprine foetuses, submitted from different points of north and central Spain, were analysed to determine the role of T. gondii in abortion and to compare the utility of the most widely used techniques in diagnosis of the congenital infection (histopathology, serology--IFAT and ELISA--and a nested-PCR). Parasite infection was diagnosed in 40 (23.1%; n = 173) ovine foetuses by at least one of the diagnostic techniques used. A higher percentage of foetuses were diagnosed using serological techniques (IFAT and ELISA) (28.3%; n = 106) than by histologic examination (8.7%; n = 173) or PCR (6.9%; n = 173). No significant association between infection and the foetal age categories was found (P > 0.05). In this study, 106 aborted foetuses were analysed by all of the three diagnostic techniques. When we compared serological results, perfect agreement between ELISA and IFAT was obtained. On the contrary, slight to fair agreements were observed when histology results were compared with those obtained by serology and PCR techniques. All the positive foetuses were aborted in the mid (60%) or last (40%) term of pregnancy, but no significant differences were found between ages of the infected and non-infected foetuses (P > 0.05). This report indicates that toxoplasmosis may be a common cause of small ruminant abortion and neonatal death in Spain and points out the necessity of using different and complementary techniques to increase the probability of detecting Toxoplasma infection in an aborted foetus.  相似文献   

4.
The aim of this study was to investigate the need for different cut-off points, according to animal age and the purpose of testing, for two of the most widely used serological techniques in bovine neosporosis, IFAT and a crude antigen ELISA (Civtest, HIPRA). Therefore, the population reference sera used were defined using a combination of multiple criteria such as epidemiological/clinical and histopathological parameters and an immunoblot test. Firstly, foetuses and breeding cattle (heifers and cows) were considered as separate subpopulations for serological evaluation. Secondly, cut-off points for each serological technique (IFAT and ELISA) according to age group (foetuses and breeding cattle) and the different practical applications (detection of infection and abortion) were calculated following the receiver operating characteristics (ROC) analysis. Cut-off points were defined, for IFAT and ELISA for aborted breeding cattle and for IFAT alone in the case of the foetuses, assuming an equivalent cost of false positive and negative results. In infected breeding cattle, for IFAT and ELISA and in foetuses for ELISA, two possible cut-off values were obtained, one for a maximum sensitivity and one for a maximum specificity and the intervals of unclear results were defined. In this case, a cut-off value for equal sensitivity and specificity was also estimated. When cut-off points for infected breeding cattle, 1:100-1:250 for IFAT and 0.306-0.451 for ELISA were applied to a target population, optimal and similar negative and positive predictive values together with similar apparent and true prevalence results were observed suggesting the possibility of using both tests interchangeably.  相似文献   

5.
Eighty foetuses from some of the main cattle-producing regions in Spain were analysed to investigate the participation of Neospora caninum in cases of bovine abortion. Diagnosis of the infection was determined by histopathological analysis complemented with immunohistochemistry, serology (IFAT and ELISA) and PCR tests. A total of 38.8% of the bovine foetuses analysed were considered to be infected by at least one of the diagnostic techniques used. Microscopic lesions consistent with Neospora infection in brain were identified in 31.3% of the samples, whereas only 10.7 and 15.3% were positive using serological and PCR analysis, respectively. Perfect agreement was shown between IFAT and ELISA, although there was little agreement among results of the other diagnostic techniques. Gestational age of aborted foetuses checked ranged from <3 to 9 months, with a mean of 5.9 months, and no difference in age was found between infected and non-infected foetuses (P>0.05). This study confirms the importance of N. caninum as a cause of abortion in Spain and underlines the need to use different diagnostic techniques to increase the chance to detect the infection in aborted foetuses.  相似文献   

6.
Babesia divergens antigen obtained from bovine and gerbil erythrocytes was compared for the diagnosis of bovine babesiosis by the indirect immunofluorescence antibody test (IFAT) and the enzyme-linked immunosorbent assay (ELISA). Using a range of sera from the field and from experimental cattle, it was found that antigen from the two sources did not differ significantly in sensitivity or specificity. Antigen from gerbils may, therefore, be used for serological surveys of bovine babesiosis caused by B. divergens. The IFAT and ELISA were also compared and it was concluded that they are equally effective in the detection of positive samples. For small-scale surveys, the IFAT using gerbil-derived antigen is adequate, but for large surveys the ELISA is preferable.  相似文献   

7.
Diagnosis of Neospora caninum infection in dogs is based on serological assays such as the indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assays (ELISA). This study evaluated two serological tests (IFAT and ELISA) for the detection of IgG antibodies to N. caninum in 300 serum samples of dogs through the optimization of cut off titers by using the two-graph receiver-operating characteristic (TG-ROC) curve. In addition, the identification of major cross-reactive antigens with Toxoplasma gondii was investigated by inhibition ELISA and immunoblotting (IB) assays. IFAT and ELISA results showed 74% agreement, with a good negative concordance (P(neg)=0.83), but a poor positive concordance (P(pos)=0.42). The great majority (86%) of sera with positive concordant results (IFAT+/ELISA+) recognized at least two out of three N. caninum immunodominant antigens, particularly the 29-32 and 35-37 kDa bands. Optimization of cut off titers in IFAT and ELISA was performed considering the reactivity to at least two out of three N. caninum immunodominant antigens as infection markers, obtaining a titer of 50 for IFAT and 200 for ELISA. Seropositivity to N. caninum was significantly associated with T. gondii-seropositive samples, particularly in ELISA (55.4%). Inhibition ELISA curves for N. caninum showed a partial heterologous inhibition, indicating some degree of cross-reactivity between N. caninum and T. gondii antigens. Inhibition IB assays showed a moderate heterologous inhibition for N. caninum antigens above 45-50 kDa. These results indicate that ELISA should be used critically when crude tachyzoite antigen preparations are employed, due to possible cross-reactivity with other related parasites as T. gondii. Also, the cut off dilution of 1:50 in IFAT showed to be the most appropriated for N. caninum serology in dogs. Therefore, we suggest that N. caninum immunodominant antigens, specially the 17 and 29-32 kDa proteins, should be selected markers in serological assays for canine neosporosis.  相似文献   

8.
Summary

Two stocks of large Babesiae from dogs originating in France, transmitted by Dermacentor reticulatus, two from North Africa, having Rhipicephalus sanguineus as vector, and one from South Africa, transmitted by Haemaphysalis leachi, were compared in cross‐immunity tests in dogs and in the indirect fluorescent antibody test (IFAT).

The French and North African stocks did not immunise against the South African one, while the North African stocks did not protect against a French one. The South African stock partially protected against a French one.

The three groups could be clearly distinguished in the IFAT These differences have practical implications for existing and future vaccines against canine babesiosis and for the serological diagnosis of atypical and chronic cases.

It is proposed to use a trinomial system of nomenclature for these groups: Babesia canis canis (Piana and Galli‐Valerio, 1895), Babesia canis vogeli Reichenow, 1937, and Babesia canis rossi (Nuttall, 1910), having Dermacentor, Rhipicephalus and Haemaphysalis ticks as their vectors respectively.  相似文献   

9.
A total of 719 serum samples collected from clinically healthy cattle from eight provinces located in different districts of South Africa were examined by the indirect enzyme-linked immunosorbent assay (ELISA) and the standard indirect fluorescent antibody test (IFAT) to determine the serological prevalence of Babesia bovis and Babesia bigemina. The results showed that 35.3% and 39.7% of cattle were positive for B. bovis and 30% and 36.5% were positive for B. bigemina antibodies on ELISA and IFAT, respectively. Mixed infections were detected in 18.2% and 26.3% of the samples using ELISA and IFAT, respectively. Consequently, the ELISAs with recombinant B. bovis spherical body protein-4 (BbSBP-4) and B. bigemina C-terminal rhoptry-associated protein-1 (BbigRAP-1/CT) were proven to be highly reliable in the serological diagnoses of bovine babesiosis in South African cattle, as evidenced by the significant concordance rates when the results were compared to those of IFAT. Moreover, the serological prevalence was significantly different among the tested provinces, in which the ranges exhibited between 15% and 73% for B. bovis infection and between 13% and 54% for B. bigemina infection. High sero-positive rates were present in Mpumalanga and KwaZulu-Natal provinces, while the lowest rate was in the North West province. Our data provide important information regarding the current seroprevalence of bovine babesiosis in South Africa, which might be beneficial in developing rational strategies for disease control and management.  相似文献   

10.
Leishmania infantum (syn. Leishmania chagasi) is the etiological agent of a widespread serious zoonotic disease that affects both humans and dogs. Prevalence and incidence of the canine infection are important parameters to determine the risk and the ways to control this reemergent zoonosis. Unfortunately, there is not a gold standard test for Leishmania infection. Our aim was to assess the operative validity of commercial tests used to detect antibodies to Leishmania in serum samples from experimental infections. Three ELISA tests (LEISCAN® Leishmania ELISA Test, INGEZIM® LEISHMANIA, and INGEZIM® LEISHMANIA VET), three immunochromatographic tests (INGEZIM® LEISHMACROM, SNAP® Leishmania, and WITNESS® Leishmania), and one IFAT were evaluated. LEISCAN® Leishmania ELISA test achieved the highest sensitivity and accuracy (both 0.98). Specificity was 1 for all tests except for IFAT. All tests but IFAT obtained a positive predictive value of 1, while the maximum negative predictive value was achieved by LEISCAN® Leishmania ELISA Test (0.93). The best positive likelihood ratio was obtained by INGEZIM® LEISHMANIA VET (30.26), while the best negative likelihood ratio was obtained by LEISCAN® Leishmania ELISA Test (0.02). The highest diagnostic odds ratio was achieved by LEISCAN® Leishmania ELISA Test (729.00). The largest area under the ROC curve was obtained by LEISCAN® Leishmania ELISA Test (0.981). Quantitative ELISA based tests performmed better than qualitative tests (“Rapid Tests”), and the test best suited to detect Leishmania in infected dogs and to provide clinically useful information was LEISCAN® Leishmania ELISA Test. This and other results point also to the need of revising the status of IFAT as a gold standard for the diagnosis of leishmaniasis.  相似文献   

11.
Objective The ability of a new commercial ELISA to detect pigs with subclinical proliferative enteropathy (PE) was compared with the traditional indirect fluorescent antibody test (IFAT). Methods Serum samples were selected from pigs with known Lawsonia intracellularis infection status and clinical signs of PE, but the sample population consisted predominantly of pigs subclinically affected by PE. Results Significant association and agreement were shown between the ELISA and IFAT assays. ELISA results correlated well with the duration of L. intracellularis shedding as detected by polymerase chain reaction. Conclusion ELISA can be successfully used to monitor L. intracellularis infection in pigs.  相似文献   

12.
Comparative serological examination of 300 serum samples from sheep slaughtered in the main abattoir in Cairo, Egypt revealed a higher prevalence of toxoplasmosis (43.7%) with the modified agglutination test (MAT), followed by the enzyme linked immune-sorbant assay (ELISA) (41.7%) and the indirect fluorescent antibody test (IFAT) (37%), while the lowest prevalence was detected with the dye test (DT) (34%). When the data from the first three serological tests were compared with that of the DT test, which was used as a reference test for toxoplasmosis, MAT had the highest sensitivity (96%), followed by ELISA (90.1%) and IFAT, which demonstrated the lowest sensitivity (80.4%). Conversely, IFAT had the highest specificity (91.4%), followed by MAT (88.9%) and ELISA (85.9%).  相似文献   

13.
Appraisal of the indirect fluorescent antibody test (IFAT) and antigen enzyme linked immunosorbent assay (ELISA) serological tests as carried out to detect cattle exposed to Theileria parva at the National Veterinary Research Centre, Muguga (NVRC), Kenya is reported. Using sera from T. parva naive cattle and cattle experimentally exposed to T. parva, the two tests were appraised in terms of their sensitivity and specificity. IFAT and ELISA had the same sensitivity of 90% while ELISA had a higher specificity (90%) than IFAT (80%). A comparison was also made of the capability of the two tests to detect exposure of dairy cattle to T. parva prior to immunization against East Coast fever (ECF). The positive outcome from the IFAT was significantly higher (chi 2 = 30.36; P < 0.001) than that from the ELISA. The agreement between the two tests was low (Kappa = 0.21). The two tests indicated a higher risk of ECF in the study area than was expected. Indications are that the ELISA has been effectively adopted at NVRC.  相似文献   

14.
Infection with the intracellular microsporidium Encephalitozoon cuniculi can cause serious disease, encephalitozoonosis, in the blue fox (Alopex lagopus). The disease diagnosis is based on clinical signs and pathological findings, and detection of E. cuniculi or circulating antibodies directed against the parasite. Indirect immunofluorescence (IFAT) and carbon immunoassay (CIA) are the most commonly used serological methods for diagnosis in this species. In the present study, an indirect ELISA (enzyme linked immunosorbent assay) was established and evaluated against IFAT by testing of 205 field samples from blue foxes. There was high agreement between the results of the ELISA and CIA (kappa=0.99), and the ELISA and IFAT (kappa=0.958). There was no significant statistical difference between the tests (p>0.05). It was concluded that the ELISA could be used to identify seropositive farmed blue foxes. The advantage of the ELISA lies in the potential of screening large numbers of animals with the goal of eradicating E. cuniculi infection in the farms.  相似文献   

15.
We carried out an inter-laboratory trial to compare the serological tests commonly used for the detection of specific Neospora caninum antibodies in cattle in Ibero-American countries. A total of eight laboratories participated from the following countries: Argentina (n = 4), Brazil (n = 1), Peru (n = 1), Mexico (n = 1), and Spain (n = 1). A blind panel of well-characterized cattle sera (n = 143) and sera representative of the target population (n = 351) was tested by seven in-house indirect fluorescent antibody tests (IFATs 1–7) and three enzyme-linked immunosorbent assays (ELISAs 1–3; two in-house and one commercial). Diagnostic performance of the serological tests was calculated and compared according to the following criteria: (1) the “Pre-test information,” which uses previous epidemiological and serological data; (2) the “Majority of tests,” which classifies a serum as positive or negative according to the results obtained by most tests evaluated. Unexpectedly, six tests showed either sensitivity (Se) or specificity (Sp) values lower than 90%. In contrast, the best tests in terms of Se, Sp, and area under the ROC curve (AUC) values were IFAT 1 and optimized ELISA 1 and ELISA 2. We evaluated a high number of IFATs, which are the most widely used tests in Ibero-America. The significant discordances observed among the tests regardless of the criteria employed hinder control programs and urge the use of a common test or with similar performances to either the optimized IFAT 1 and ELISAs 1 and 2.  相似文献   

16.
The aim of this study was to compare two serologic tests used to support a diagnosis of equine protozoal myeloencephalitis (EPM). Serum and cerebrospinal fluid (CSF) samples were analyzed for antibodies to Sarcocystis neurona and Neospora hughesi by indirect fluorescent antibody testing (IFAT) and surface antigens of S. neurona and N. hughesi by enzyme-linked immunosorbent assay (ELISA). The samples originated from neurologic horses with confirmed and suspected EPM (nine S. neurona, three N. hughesi), from neurologic horses with confirmed neurologic diseases other than EPM (16 horses) and from healthy horses (10). The IFAT on CSF and ELISA titer ratios showed equal sensitivity in diagnosing EPM caused by S. neurona. The ELISA titer ratios showed slightly greater specificity in diagnosing EPM than the IFAT on CSF. Overall agreement between the IFAT on CSF and ELISA titer ratio was 90.9%. The IFAT on CSF and ELISA serum/CSF ratio are indicated to help support a laboratory diagnosis of EPM.  相似文献   

17.
Bovine neosporosis is an important disease in dairy herds in Argentina. Embryo transfer (ET) technology offers the opportunity to minimize vertical transmission of neosporosis. The objective of this work was to compare the serological status and the incidence of abortion between heifers obtained via natural service and heifers obtained by ET from Neospora‐infected cows. Progeny from eight Neospora‐infected cows belonging to a commercial dairy farm where ET was routinely performed were studied. Between 1996 and 2001, 16 and 15 heifers were obtained by natural service and ET, respectively. Serostatus and reproductive performance were recorded in both progeny. Serostatus was determined by indirect fluorescent antibody test (IFAT). All heifers obtained by natural service were seropositive and six of 16 pregnancies ended in abortion. All heifers obtained by ET were seronegative and only two abortions of 15 pregnancies were recorded.  相似文献   

18.

Background

Lawsonia intracellularis is a common cause of chronic diarrhoea and poor performance in young growing pigs. Diagnosis of this obligate intracellular bacterium is based on the demonstration of the microbe or microbial DNA in tissue specimens or faecal samples, or the demonstration of L. intracellularis-specific antibodies in sera. The aim of the present study was to evaluate a blocking ELISA in the detection of serum antibodies to L. intracellularis, by comparison to the previously widely used immunofluorescent antibody test (IFAT).

Methods

Sera were collected from 176 pigs aged 8-12 weeks originating from 24 herds with or without problems with diarrhoea and poor performance in young growing pigs. Sera were analyzed by the blocking ELISA and by IFAT. Bayesian modelling techniques were used to account for the absence of a gold standard test and the results of the blocking ELISA was modelled against the IFAT test with a "2 dependent tests, 2 populations, no gold standard" model.

Results

At the finally selected cut-off value of percent inhibition (PI) 35, the diagnostic sensitivity of the blocking ELISA was 72% and the diagnostic specificity was 93%. The positive predictive value was 0.82 and the negative predictive value was 0.89, at the observed prevalence of 33.5%.

Conclusion

The sensitivity and specificity as evaluated by Bayesian statistic techniques differed from that previously reported. Properties of diagnostic tests may well vary between countries, laboratories and among populations of animals. In the absence of a true gold standard, the importance of validating new methods by appropriate statistical methods and with respect to the target population must be emphasized.  相似文献   

19.
Two rapid tests evaluated in dogs considered to be of high risk of infection with the Chagas parasite Trypanosoma cruzi using two immunochromatographic assays: Trypanosoma Detect™ for canine, InBios, Seattle, WA and CHAGAS STAT-PAK™ assay, Chembio Diagnostic Systems, Medford, NY, in south central Louisiana. For this purpose a serological survey was carried out in a total of 122 dogs and a serum bank was created. These 122 animals were first tested by IFAT that was used as the standard test. From the serum bank 50 samples were tested using the two rapid Chagas assays and results compared to the standard test IFAT. The serological survey using IFAT showed a prevalence of T. cruzi infection in 22.1% of the tested dogs. In the immunochromatographic assays, 13 and 11 animals were positive on rapid assay: Trypanosoma Detect™ for canine, InBios and CHAGAS STAT-PAK™, Chembio Diagnostic Systems, respectively compared to 11 positive by IFAT. These two immunochromatographic tests have shown high susceptibility and specificity compared to our standard method IFAT. The rapid, easy and accurate screening assays used in conjunction with confirmatory tests, would be an excellent tool for veterinarians to diagnose T. cruzi infection. Early detection of T. cruzi infection may prevent complications through an effective treatment. Greater awareness by veterinarians of the risk, clinical findings, history along with diagnostic methods will contribute greatly to an understanding of the true prevalence of Chagas disease in dogs in Louisiana.  相似文献   

20.
Human visceral leishmaniasis is endemic in the northeast of Brazil, where the domestic dog is an important parasite reservoir in the infectious cycle of Leishmania chagasi. In this study, we evaluated the clinical signs of canine visceral leishmaniasis (CVL), serum protein profile and the antileishmanial IgG antibody production in 86 dogs living in northeast endemic areas of leishmaniasis. Thirty dogs from a leishmaniasis-free area were used as a control group. The major clinical signs of CVL seen were emaciation and skin ulcers (80%), followed by onychogryphosis and conjunctivitis (73%). Depilation was observed in 60% of animals while lymphadenomegaly, splenomegaly, liver enlargement or kidney involvement was less frequent (< or =20%). VL seropositive dogs presented with serum hyperproteinemia, hypoalbuminemia, hypergammaglobulinemia and decreased albumin/globulin ratio. A lower sensitivity and higher specificity was observed for promastigote indirect fluorescent antibody test (IFAT) (83 and 100%, respectively) compared with enzyme-linked immunosorbent assay (ELISA) (94 and 90%), which uses a crude extract of Leishmania. There was a positive correlation between IFAT and ELISA titers of antileishmanial IgG antibodies (Spearman test, P < 0.05), which was augmented in CVL dogs. This study found that the determination of serum protein, A/G ratio and the use of two different leishmanial serological tests like IFAT and ELISA are essential in CVL screening.  相似文献   

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