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1.
A monoclonal blocking enzyme-linked immunosorbent assay (ELISA) and an indirect haemagglutination assay (IHA) were applied to serum samples from 124 specific pathogen-free (SPF) breeding and multiplying herds, which participate in the routine serological surveillance of the Danish SPF programme. Clinical and pathological observations of the herds and microbiological culturing of Mycoplasma hyopneumoniae were used to calculate herd sensitivity, herd specificity and herd predictive values for the two serological assays. The ELISA was superior to the IHA in herd sensitivity and herd specificity, with values of 93 per cent and 96 per cent, respectively, for the ELISA, and 61 per cent and 92 per cent for the IHA. During the six month period of evaluation 2.5 per cent of the herds were infected with M hyopneumoniae each month. At this level the IHA was found to have a positive herd predictive value of 16 per cent, compared with 39 per cent for the ELISA. The negative herd-predictive value on the same level was 99.8 per cent for the ELISA and 98.9 per cent for the IHA. If the assays were applied to a group of herds with a herd prevalence of M hyopneumoniae infection of 30 per cent (as is the case with the production herds in the Danish SPF programme) the predictive value of a positive herd diagnosis would be 91 per cent for the ELISA and 76 per cent for the IHA, and the predictive value of a negative herd diagnosis would be 97 per cent with the ELISA and 85 per cent with the IHA.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

2.
Measurements of the conductivity of quarter milk samples were made in 31 cows in a 70-cow herd in southeast England, for a period of 15 weeks. Over this period, 42 per cent of cow-weeks and 20 per cent of quarter-weeks had an increase in quarter milk conductivity of 10 per cent of more compared with the mean conductivity of the previous 14 milkings. Fourteen per cent of quarter-weeks had an increase in conductivity of 15 per cent or more. The geometric mean somatic cell count (cell count) was higher in quarter-weeks with a 10 per cent or greater increase in conductivity than in quarter-weeks with a conductivity change of less than 10 per cent. At a conductivity threshold of 10 or 15 per cent and a cell count threshold of 200,000 or 400,000 cells/ml the specificity of this system was estimated to be 85 to 92 per cent, the sensitivity 40 to 54 per cent, the negative predictive value 87 to 93 per cent and the positive predictive value 33 to 55 per cent. The positive predictive value of the individual quarter milk conductivity was insufficiently accurate to be used as the sole criterion for the selection of quarters for early antibiotic treatment.  相似文献   

3.
The diagnostic performance of an ELISA for the detection of antibodies to the small ruminant lentiviruses (SRLVs) maedi-visna virus and caprine arthritis-encephalitis virus in milk and corresponding blood samples was evaluated in 50 sheep. The agreement between ELISA results in blood and milk was 90 per cent, and the κ value was 0.79. In addition, a serological survey in the central zone of Spain was performed using milk samples from 413 animals (250 sheep and 163 goats) from 12 flocks/herds. All flocks/herds had some animals that were positive for SRLV. Among the animals, 60.0 per cent of the sheep and 8.0 per cent of the goats tested were seropositive. Each sample was also tested using a PCR technique, which increased the percentage of positive animals detected. Using a combination of ELISA and PCR gave a total of 72.2 per cent of sheep and 28.8 per cent of goats positive for SRLV.  相似文献   

4.
The performances of a competitive enzyme-linked immunosorbent assay (ELISA) using a group specific monoclonal antibody against bluetongue virus, an indirect ELISA and the standard agar gel immunodiffusion (AGID) test were compared in the detection of serum antibody against bluetongue virus. Test sera consisted of 1300 bovine, 530 ovine and 160 carpine samples from bluetongue-free areas of Canada, 605 bovine and ovine field samples from the USA and Barbados and 464 samples from 79 cattle and sheep experimentally infected with 19 South African and five USA serotypes of bluetongue virus. The diagnostic specificity of the competitive ELISA, as determined for the bluetongue virus-free cattle sera was superior (99.92 per cent) to that of the indirect ELISA (99.85 per cent) and the AGID (99.0 per cent). The specificities of the competitive ELISA for sheep (99.63 per cent) and goats (100.0 per cent) sera were also higher than those of the AGID test. The performance of the ELISA tests was similar whether a gamma-ray-irradiated (2.0 Mrad) or a non-irradiated bluetongue virus antigen preparation was used. The competitive ELISA results for bovine field sera from endemic areas demonstrated a relatively low level of agreement (92.04 per cent) with AGID test results, with 9.7 per cent false negatives. The possible presence in these sera of antibody to cross-reacting antigens or to other orbiviruses, eg, epizootic haemorrhagic disease virus, which react in the AGID but not in the competitive ELISA may account for this lack of agreement.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

5.
Echinococcus granulosus is an important zoonotic infection of dogs. The purpose of the present study assessed the performance of two laboratory diagnostic methods with arecoline purgation and necropsy in infected dogs. In total 65 dogs were successfully experimentally infected with protoscoleces of E. granulosus from ovine infection. At 14-34 days post-infection groups of dogs were purged with arecoline hydrobromide and then necropsied. Faecal samples were tested at weekly intervals by coproantigen ELISA and coproPCR. The necropsy infection rate with E. granulosus was 89.2 per cent. Only 43 per cent of dogs were successfully purged after one arecoline dose; this percentage increased to 76.9% for two doses of arecoline purgation. E. granulosus coproantigen was detected by coproELISA in 82.8% of faeces. The positive and negative predictive values for coproantigen ELISA were 96 and 44.4% respectively. E. granulosus DNA was detected in pre-patent faecal samples by coproPCR in 25.9% of dogs. These results indicate that coproELISA is more sensitive than arecoline purgation for the detection of pre-patent E. granulosus infection in dogs. CoproPCR detected E. granulosus DNA in dog faeces by 21 days post-infection before egg production.  相似文献   

6.
An ELISA developed to diagnose Fasciola hepatica infection in cattle by detecting serum antibodies was adapted and validated for use with samples of bulk-tank milk. The prevalence of the infection in 61 dairy herds was established by using serum antibody levels or faecal egg counts measured in a proportion of the cows in each herd. The correlation between the results of the ELISA and the herd seroprevalence was 0.83. Using a cut-off value of 27 per cent positive, the bulk-tank ELISA identified herds in which more than 25 per cent of the cows were infected with a diagnostic sensitivity of 96 per cent (95 per cent confidence interval 89 to 100 per cent) and a diagnostic specificity of 80 per cent (95 per cent confidence interval 66 to 94 per cent). By applying the ELISA to 623 herds in England and 445 herds in Wales, the prevalence of F hepatica infection in England was estimated to be 48 per cent (95 per cent confidence interval 46 to 54 per cent), and in Wales 86 per cent (95 per cent confidence interval 84 to 90 per cent).  相似文献   

7.
A recombinant LipL 32 antigen-based dipstick ELISA was developed as a screening test for the detection of leptospiral antibodies in serum samples from dogs. The antibodies were detected by a change in the colour of the substrate solution when the recombinant antigen-coated dipsticks were dipped into it. The relative sensitivity, specificity and accuracy of the test, compared with the standard microscopic agglutination test, were 95.9 per cent, 93.8 per cent and 94.8 per cent, respectively.  相似文献   

8.
A comparison was made between an indirect enzyme-linked immunosorbent assay (ELISA) and the virus neutralisation test (VNT) for the detection of antibodies to rinderpest virus in field sera. The results did not agree for 6 per cent of the sera tested where 3 per cent of the samples gave ELISA positive/VNT negative and 3 per cent gave ELISA negative/VNT positive. The latter sera all had high levels of IgM antibody, which may indicate animals being at an early stage of infection or detection of a non-specific reaction. The ELISA results give a representative picture of the immune status for field surveys and a greater number of sera can be assayed with relative ease, compared to the traditional serum neutralisation test.  相似文献   

9.
The present study was designed to evaluate a commercial ELISA kit (Institut Pourquier) for the diagnosis of ovine and caprine paratuberculosis under Australian conditions and to compare its accuracy with the existing AGID test. The sensitivity of the ELISA in sheep and goats was 34.9% and 56.4%, with a specificity of 98.8% and 100.0%, respectively. Sensitivity of AGID was 13.8% for sheep and 39.5% for goats, with specificity of 100.0% for both species. The sensitivity of the ELISA in sheep depended on the category of histological lesions. AGID and ELISA were conditionally independent, and appeared to detect overlapping but distinct subgroups of infected animals. The ELISA was significantly more sensitive than the AGID. The ELISA was simple to perform, robust and repeatable. Coefficients of variation of <12.0% were observed for positive and negative controls included on 193 plates over a 10-month period and there was a high level of intraassay repeatability with 12.0% of the duplicate samples having CV of >15.0%.  相似文献   

10.
In 2007, a human Q fever epidemic started, mainly in the south eastern part of The Netherlands with a suspected indirect relation to dairy goats, and, to a lesser degree, to dairy sheep. This article describes the Q fever prevalences in Dutch dairy goat and dairy sheep bulk tank milk (BTM) samples, using a real-time (RT) PCR and ELISA. Results of BTM PCR and ELISA were compared with the serological status of individual animals, and correlations with a history of Q fever abortion were determined. When compared with ELISA results, the optimal cut-off value for the RT-PCR was 100 bacteria/ml. In 2008, there were 392 farms with more than 200 dairy goats, of which 292 submitted a BTM sample. Of these samples, 96 (32.9 per cent) were PCR positive and 87 (29.8 per cent) were ELISA positive. All farms with a history of Q fever abortion (n=17) were ELISA positive, 16 out of 17 were also PCR positive. BTM PCR or ELISA positive farms had significantly higher within-herd seroprevalences than BTM negative farms. In the south eastern provinces, the area where the human Q fever outbreak started in 2007, a significantly larger proportion of the BTM samples was PCR and ELISA positive compared to the rest of The Netherlands. None of the BTM samples from dairy sheep farms (n=16) were PCR positive but three of these farms were ELISA positive. The higher percentage of BTM positive farms in the area where the human Q fever outbreak started, supports the suspected relation between human cases and infected dairy goat farms.  相似文献   

11.
Percutaneous fine-needle biopsy was used to investigate thoracic and abdominal masses in the dog and cat. One hundred and thirty-two cases were included in the study; 20 cases were excluded from the comparative study due to poor cellularity or blood contamination (retrieval rate 86.8 per cent). One hundred samples (56 dogs and 44 cats) were classified by cytology as neoplastic. All the cytological diagnoses of neoplasia were confirmed by histological samples obtained either by non-surgical methods, at surgery or during postmortem examination. No false positive diagnoses of neoplasia were made. Thirty-two samples were cytologically classified as 'negative for neoplasia'. Subsequent histological examination revealed 18 true negative and 14 false negative results. The procedure had an overall 89.4 per cent (118 cases out of 132) agreement between the diagnosis of inflammatory disease versus neoplasia, with a sensitivity of 87.8 per cent, a specificity of 100 per cent, a predictive value of a positive test of 100 per cent and a predictive value of a negative test of 56.3 per cent.  相似文献   

12.
Brush cytology was used as a diagnostic aid in 85 cats affected with chronic intranasal disease. Fifty-three of these cases, sampled over a five-year period, were included in this study, while the other cases were excluded due to poor cellularity of the cytological samples (nine cases) or a lack of histological or follow-up data (23 cases). Thirty-six brush samples were classified by cytology as inflammatory. Subsequent histological examination revealed a false negative diagnosis of neoplasia in six cats, two of which had malignant tumours (one adenocarcinoma and one lymphoma), the remaining four having benign tumours (two adenomas and two osteochondromas). Seventeen samples were classified by brush cytology as neoplastic. This was confirmed in 16 of these cases by histology or follow-up (nine epithelial malignant tumours, six lymphomas and one osteosarcoma). In the remaining case, a false positive diagnosis of lymphoma was made. The procedure had an overall 86.8 per cent (46/53) agreement between the diagnosis of inflammatory conditions versus neoplasia, with a sensitivity of 72.7 per cent, a specificity of 96.8 per cent, a predictive value of a positive test of 94.1 per cent and a predictive value of a negative test of 83.3 per cent.  相似文献   

13.
Serological surveys were carried out to determine the prevalence of pestiviral infections in sheep and pigs in Northern Ireland. Sera from 918 ewes in 92 flocks from 10 regions were tested by ELISA for antibodies to border disease virus and positive results were obtained from 49 ewes (5.3 per cent) in 28 flocks (30.4 per cent). There were highly significant geographical variations in its flock prevalence ranging from 0 per cent in the Enniskillen region to 70 per cent in the Coleraine region. There was no significant association between the proportion of seropositive flocks and the presence of cattle on the farm (P = 0.583). In the positive flocks, the average rate of seroprevalence was 17.5 per cent, and the highest was 40 per cent. Comparative neutralisation studies on 14 positive sera with bovine viral diarrhoea virus type I (BVDV I) and border disease virus revealed higher titres (> or = four-fold) to BVDV I in all cases. Only one positive result was obtained when fluids from 186 aborted ovine fetuses were tested for border disease virus by ELISA. Serum samples from 680 pigs in 46 herds were tested for virus neutralising antibodies to border disease virus. Twenty sera (2.9 per cent) were cytotoxic, and only one of the remaining 660 sera gave a positive result. This sample tested negative for classical swine fever by ELISA, and comparative neutralisation studies showed that it had a four-fold higher titre to BVDV I than to border disease virus.  相似文献   

14.
The significant economical consequences of infections with Ascaris suum in pigs are already well documented. However, due to the subclinical nature of the disease and the lack of practical diagnostic means, ascariasis often remains undiagnosed. Here we describe the development and evaluation of a novel indirect ELISA using the purified A. suum haemoglobin (AsHb) molecule as an antigen. Initial validation using sera from 190 pigs experimentally infected twice a week with A. suum and Trichuris suis (25 and 5eggskg(-1)day(-1) respectively) demonstrated that the AsHb ELISA is able to detect long-term exposure to A. suum with a high sensitivity and specificity (99.5% and 100.0% respectively). Furthermore, this serological technique proved to be more sensitive than faecal examination on week 7 and 14 of the experiment (99.5% and 100% compared to 59.5% and 68.4% respectively). Cross-reactivity caused by T. suis infection was shown to be limited after analysing sera from pigs with an experimental T. suis mono-infection. Seroconversion was shown to occur from week 6 onwards in pigs receiving 100 A. suum eggs 5 times a week. Preliminary testing of the ELISA on six randomly selected farms confirmed the results obtained in the artificial infection trials, showing a higher sensitivity of the serologic method compared to faecal examination. Finally, the ELISA was used to investigate Ascaris infection rates on 101 conventional Flemish pig farms. The results showed that on 38.6% of the farms less than 20% of the tested samples were seropositive, while in 19.8% of the farms 80-100% of all pigs were seropositive. The results of this study suggest that the AsHb ELISA could provide pig farmers and veterinarians with an easier and more sensitive way to estimate the overall prevalence of A. suum on their farm.  相似文献   

15.
Recombinant p40 produced by baculovirus was used in an ELISA to screen samples of serum taken from 80 cats in Istanbul. The sera were also analysed for feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV). Antibodies to Borna disease virus- (BDV) p40 were detected in 34 (42-5 per cent) of the 80 cats. Seventy-three per cent of the sera which were positive for FIV and 26 per cent of the sera which were negative for FIV had antibodies to BDV. There was no difference in the percentage of sera which were positive for BDV between the cats that were positive or negative for FeLV. Three of the cats had neurological disease and two of these had antibodies to BDV. Six sera with low, medium or high optical densities (ODS) by ELISA were analysed by Western blotting. Only the sera with medium and high ODS reacted specifically with p40 at a dilution of 1 in 1,000.  相似文献   

16.
Objectives : To determine the reliability of early passive hip laxity examination in predicting moderate and severe hip dysplasia. Methods : The passive hip laxity of 104 Estrela Mountain dogs was evaluated using the Ortolani method, conventional hip‐extended radiography and the PennHIP method, when the animals were between four and 12 months of age. After the age of one, dogs were re‐examined for hip dysplasia using the Fédération Cynologique Internationale scoring system. The passive hip laxity results were subsequently compared with the Fédération Cynologique Internationale scores for sensitivity, specificity, positive predictive value, negative predictive value and accuracy. Results : The PennHIP method achieved the best results in specificity (93 per cent), positive predictive value (91 per cent) and accuracy (88 per cent), and the Ortolani method was the best in sensitivity (92 per cent) and negative predictive value (92 per cent). Clinical Significance : The Fédération Cynologique Internationale grades D and E can be predicted with confidence even before animals are one year old.  相似文献   

17.
Serially collected epithelial samples from lesions in the mouth and on the feet of calves experimentally infected with foot-and-mouth disease (FMD) type O1 BFS 1860 were assayed for the presence of FMD viral antigen using a double antibody sandwich enzyme-linked immunosorbent assay (ELISA) and a complement fixation (CF) test. The amount of infectious virus in each sample was also determined. FMD viral antigen was detected by ELISA in 70 per cent of the mouth samples and 92 per cent of samples from the feet. The CF test was less sensitive; it detected antigen in 44 per cent of mouth and 85 per cent of foot samples. In mouth samples the amount of antigen decreased rapidly becoming undetectable by the fourth day of sampling whereas in foot samples the quantity of antigen declined more slowly, and could be detected until the seventh day of sampling. Therefore it was concluded that the age of lesion and the site from which epithelial samples are collected are both important determinants in the laboratory diagnosis of FMD. In cattle, foot lesions are more likely than mouth lesions to yield antigen and to remain positive for a longer period.  相似文献   

18.
An indirect micro enzyme-linked immunosorbent assay (ELISA) for detecting antibodies to Aujeszky's disease virus in pigs is described. A control antigen prepared from infected cells was included for each serum tested. Of 243 sera from serologically positive farms, 175 (72 per cent) and 147 (60 per cent) were positive by the ELISA test and microtitre serum neutralisation test, respectively. Failure to include a control antigen for each serum would have resulted in 14 sera (6 per cent) being differently recorded. Results for sera from experimental and field infections indicated that seroconversion was more quickly detected by the ELISA test than the microtitre serum neutralisation test. In addition to greater sensitivity the ELISA test has other advantages over the serum neutralisation test. ELISA is a rapid, cheap test which is not dependent on a continuous supply of cell cultures and which can be readily automated.  相似文献   

19.
Farmers recorded the clinical signs of cows with clinical mastitis and submitted milk samples for bacteriological examination, so that the clinical signs could be correlated with the bacteriological findings. Odds ratios for the demeanour of the cow, the appearance of the milk, milk yield, udder texture, and the administration of parenteral antibiotics were calculated for mastitis cases classified in terms of their microbiology as either enterobacteriaceae, major Gram-positive pathogens, minor pathogens, 'no growths' or 'all other pathogens'. Animals infected with enterobacteriaceae had the highest odds of being reported as having a reduced milk yield, swollen or hard udders, watery milk and/or being systemically sick. A logistic regression model was used to predict the Gram-staining characteristics of the bacteria causing clinical mastitis. The clinical findings found to be significant predictors in the model were the demeanour of the cow and its milk yield. The regression model was used as a basis for a predictive test. Using a test data set, the sensitivity of the test was 28 per cent, its specificity was 96 per cent, the positive predictive value was 74 per cent and the negative predictive value was 80 per cent. The overall accuracy of these predictions was 79 per cent.  相似文献   

20.
OBJECTIVE: To evaluate the efficacy of a novel monoclonal antibody (MAb)-based dot-blot ELISA for detection of Leptospira antigens in urine samples of cattle. SAMPLE POPULATION: Blood and urine samples of 45 test cattle from 5 farms in Chonburi province and 20 control cattle from 2 farms in Khon Kaen province in Thailand. PROCEDURE: Blood and urine samples were assayed (microscopic agglutination test and urine antigen test) for Leptospira infection by use of an MAb-based dot-blot ELISA, and results for the ELISA were compared with those for dark-field microscopy (DFM), microbial culture, and a polymerase chain reaction (PCR) assay. RESULTS: All urine samples with positive results for DFM, microbial culture, PCR assay, or > 1 of these tests also had positive results when tested by use of the MAb-based dot-blot ELISA, except for 1 sample that had positive results only for the PCR assay. Detection limits of the dot-blot ELISA were 10(3) leptospires/mL of urine and 9.3 ng of Leptospira homogenate. Comparison revealed that the diagnostic sensitivity, specificity, efficacy (accuracy), positive predictive value, and negative predictive value for the ELISA were in agreement with results for DFM (100%, 72.72%, 80%, 57.14%, and 100%, respectively), microbial culture (100%, 61.54%, 66.62%, 28.57%, and 100%, respectively), and PCR assay (95.45%, 100%, 91.77%, 100%, and 95.83%, respectively). CONCLUSIONS AND CLINICAL RELEVANCE: The MAb-based dot-blot ELISA is suitable as a tool for detecting leptospires in urine samples of cattle.  相似文献   

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