牛布鲁菌病补体结合试验测量不确定度评估

本试验应用补体结合试验方法对牛布鲁菌病进行检测,分别从溶血素效价测定、补体效价测定、抗原效价测定及血清效价测定4个方面分析了补体结合试验测量不确定度来源,并计算了合成不确定度、扩展不确定度,最终样品测量结果报告值为:X=(1.06±0.043) mg/mL(95％置信区间),即本试验中判定的可以抑制50％溶血的血清最高效价所对应的溶血素浓度为1.06mg/mL,不确定度U为0.043.     

An intradermal test for the diagnosis of brucellosis in extensively managed cattle herds

The application of a delayed hypersensitivity test for the diagnosis of bovine brucellosis was examined in a series of field experiments. The test is based on the intradermal injection of ‘Brucellin’, a lipopolysaccharide-free protein extract of Brucella abortus strain 45/20.

The Brucellin test was compared with the complement fixation (CF) test in 8656 cows of mixed age and known vaccination and herd status. An intradermal injection of 0.1 ml of the allergen was made in either the cervical region or the caudal fold. The injection site was examined 72 h after administration of Brucellin and any increase in skin thickness of 2 mm was regarded as positive.

When administered into the caudal fold site the Brucellin test had a sensitivity relative to the CF test of between 52 ± 14% and 61 ± 6%. The relative specificity of the test exceeded 99%.

Calfhood vaccination with B. abortus strain 19 did not result in positive Brucellin test results. There was no evidence that the injection of Brucellin induced a serological response.

Despite the low relative sensitivity of the Brucellin test, it is a useful low-cost tool for identifying infected herds. It is a least as effective as slaughterhouse surveillance systems.     

The detection of bovine herpesvirus type 1 (BHV 1) using an intradermal test. II. Experimental studies

A purified concentrated BHV1 antigen that had been tested in field trials was also tested under isolated conditions in BHV1 positive and negative cattle. The antigen proved to act specifically even after storage for 2 years at +4 degrees C or for 6 months at 37 degrees C. The best results were obtained when the test was read 48-72 hours after the injection, which is in agreement with the field trials. The increase in skin thickness decreased unless boosted by infection or vaccination gradually. The test is unsuitable to control a vaccination programme. There was no correlation between the increase in thickness and humoral antibody titers. Repeated application of the intradermal injection of the antigen did not result in seroconversion in seronegative cattle. The biological limits of the test evaluation are discussed.     

The detection of bovine herpesvirus type 1 (BHV 1) using an intradermal test. I. Field studies

An intradermal test (delayed hypersensitivity test) for the diagnosis of BHV1 infection was evaluated in 791 cattle of 16 dairy farms. The skin reactions were compared with the results of serological examinations using a commercial BHV1 ELISA kit (Trachitest). As antigen concentrated, purified and inactivated BHV1 was used. The skin reaction (increase of the skin fold thickness) was used for the interpretation of test results. The best results were obtained with the control of the skin reaction on the third day after injection of the antigen. From 393 serologically BHV1 negative cattle with an age of more than 6 months 391 (99.5%) had a skin reaction up to 1.0 mm and 2 animals (0.5%) had a reaction of 1.3 and 1.9 mm, respectively. The mean increase of skin fold thickness was 0.2 mm. Out of 291 serologically BHV1 positive cattle with an age of more than 6 months 270 had antibodies from natural infection and, partially, from additional vaccination with inactivated BHV1 vaccine. 266 (98.5%) of these animals showed a skin reaction of more than 2.0 mm, in 3 animals (1.1%) a skin reaction up to 1.0 mm was observed and 1 animal (0.4%) had a reaction of 2.0 mm. The mean increase of the skin fold thickness was 6.3 mm. 21 animals had BHV1 antibodies only because of vaccination with inactivated BHV1 vaccine. Only 4 animals had a skin reaction of more than 2.0 mm. Among 107 animals with an age up to 6 months 30 were serologically BHV1 positive and 77 were BHV1 negative. In all animals the skin reaction was less than 1.0 mm, the mean was 0.2 mm.     

The vaginal mucus agglutination test in the diagnosis of bovine brucellosis

Of 1140 vaginal mucus agglutination tests (VMAT) on specimens obtained in 1971-72 from 663 dairy cows in seven herds infected with brucellosis, 97 were positive. When the VMAT was positive one or more serological tests were also positive. Of the 97 corresponding serum agglutination tests 80 sera had titres of more than 533 international units. Only 69.8 per cent of VMAT from serologically positive cows were positive. No evidence was found of non-specific agglutinins in vaginal mucus and positive VMAT reactions appeared to be specific for field infection. Three cows showed evidence of local agglutinins in the vagina. Hence herd testing by VMAT has no advantage over tests of blood serum but the test could be an aid in establishing whether individual cattle are infected.     

Field evaluation of the single intradermal cervical tuberculin test and the interferon-gamma assay for detection and eradication of bovine tuberculosis in Spain

A field comparison of the interferon-gamma (IFN-gamma) assay and the single intradermal cervical tuberculin (SICT) test for the diagnosis of bovine tuberculosis was conducted. A total of 1136 cattle belonging to 85 herds placed in 'Castilla y León' (northwestern Spain) were chosen, and 21 of these herds were subjected to the diagnostic assays two or three times at intervals of at least 4 months. All the animals positive to any of the tests were slaughtered and tuberculosis was confirmed by culture isolation method (CIM) and further identification by means of PCR. Only 10.6% of cattle reacted with the bovine PPD in the SICT test, a percentage that increased to 12.8% in the IFN-gamma assay. The sensitivity of the IFN-gamma assay compared to CIM was shown to be higher (84.9%) than that of the SICT test (80.2%), but the combination of both tests offered the highest sensitivity (92.9%). The number of false positive reactors (those animals in which CIM was negative) was considerably higher for the IFN-gamma assay than for the SICT test and, conversely, the number of false negative animals (M. bovis isolation but negative immunological result) was higher for the skin test than for the interferon assay. In the herds tested twice, tuberculosis was eradicated after the second cycle of testing in 50%, and in 75% after the third cycle in herds tested three times. The combination of these two techniques instead of separately seems, therefore, to be useful in eradication programmes against bovine tuberculosis.     

应用斑点酶联免疫试验快速检测绵羊布鲁氏菌病

Dot- ELISA首先为 Hankes和 Harbrink等建立。Pappas等正式称其为 Dot- ELISA。鉴于本方法无需复杂设备 ,目前已作为一种免疫酶技术在医学领域广泛应用。将本技术应用于绵羊布鲁氏菌( Brucella ovis)病的诊断 ,目前尚未见资料报道。本文旨在建立本学科的试验方法 ,并证明其方法的可靠性和可行性。1　材料与方法1 .1　载体　混合纤维素酶微孔滤膜 (上海产 ,批号870 82 9)。1 .2　试剂　 Br.ovis菌体抗原、热酚抗原、超声提取抗原、碱提取抗原均为新疆流研所刘志文制备。酶标记兔抗羊 Ig G由上海生物制品研究所提供。1 .3　制膜　取前…     

Diagnosis of bovine brucellosis by skin test: conditions for the test and evaluation of its performance.

Brucellergene OCB (Rh?ne-Mérieux) was used as an allergen to define the intrinsic parameters of a skin test and to compare its properties with serology for the diagnosis of bovine brucellosis. The skin test was also evaluated for its capacity to solve problems associated with false positive reactions in serological tests. The optimal reading delay for the skin test was 72 hours. The brucellosis allergic reaction was two to three times less intense than the tuberculosis allergic reaction. An increase of 1.1 mm or more in the skin thickness was therefore considered to be an adequate cut-off. The specificity calculated for 1192 brucellosis-free animals (including animals from brucellosis-free herds in which false positive serological reactions had been reported) was 99-83 per cent (95 per cent confidence interval [CI] 99-40 to 99-98 per cent). The sensitivity determined from 27 experimentally infected heifers ranged from 93 per cent (95 per cent CI 76 to 100 per cent) to 78 per cent (95 per cent CI 58 to 91 per cent) when measured respectively one and six months after the infection. Allergic reactions could be detected in vaccinated animals up to four-and-a-half years after the vaccination. On the other hand, no sensitisation was recorded in na?ve animals after up to eight monthly injections of the allergen. The skin test gave valuable information, in combination with the serological tests, in both acute and chronic brucellosis. The skin test discriminated brucellosis clearly from false positive serological reactions due to infections with Yersinia enterocolitica O9.