两种方法检测奶牛结核病的比较分析

采用常规皮内变态反应(TST方法)对唐山、秦皇岛、沧州和石家庄四个市共计2 204头奶牛进行牛结核病检测,采用IFN-γ试验方法对PPD检测结果为阳性、可疑的牛以及部分阴性牛进行复检。将IFN-γ试验与TST检测结果相比较,其敏感性为78.26%,特异性为89.09%,符合率为84.33%。研究表明单独使用PPD皮内变态反应方法在牛结核病的诊断方面存在特异性差的缺点,用IFN-γ试验对TST试验阳性和可疑牛进行复检可以提高特异性,避免造成假阳性牛的误杀;该方法在临床上适合推广应用。     

田阳县应用皮内变态反应监测牛结核病

PPD(结核菌素纯蛋白衍生物)颈部皮内变态反应法是国家检测结核病的法定方法,我县在组织开展牛结核病PPD检测方法及迟缓型变态反应时,准确把握PPD检测法和皮内变态反应方法,现将临床应用和结果进行分析,供同行参考。     

不同方法检测奶牛结核病的结果分析

采用常规皮内变态反应(TST)、牛全血干扰素试验方法(IFN-γ)及胶体金3种检测方法,对2个奶牛场的600头奶牛进行结核病检测,发现结核病阳性率均偏高,表明奶牛场结核病感染较严重。将3种方法检测结果的敏感性、特异性及符合率进行比较,结果表明:单独使用PPD皮内变态反应方法或胶体金方法在牛结核病的诊断方面存在特异性及符合率差的缺点;用IFN-γ实验进行检测,可以提高特异性,避免因假阳性造成的误杀,适合在本地区推广应用。     

两种奶牛结核病检测方法的比较

本研究中采用常规皮内变态反应(TST)对广西南宁、柳州共计2 818头黑白花奶牛进行牛结核病检测,采用抗原特异性IFN-γ试验方法对PPD检测结果为阳性、可疑的牛以及部分阴性牛进行复检。将抗原特异性IFN-γ试验与TST检测结果相比,其敏感性为53.33%,特异性为70.49%,符合率为67.11%。本研究证实单独使用PPD皮内变态反应方法在牛结核病的诊断方面存在非特异性强等缺点,易造成假阳性牛的误杀;用抗原特异性IFN-γ试验对TST试验阳性和可疑牛进行复检可以提高特异性,该方法在临床上有重要应用价值。     

牛结核病污染场4种检测方法评估

为寻找最适合牛结核病污染场的检测方法,加快牛结核病净化,采用牛结核菌素(PPD)皮内变态反应试验(TST)、比较皮内变态反应试验(SICTT)、牛结核病ELISA-γ-干扰素试验(IFN-γ-ELISA)、牛结核病抗体ELISA试验4种方法,对某结核病污染牛场300头奶牛进行检测,并以TST为参考标准,对不同检测方法的检测结果进行对比分析。结果显示,这4种方法中,IFN-γ-ELISA的阳性检出率最高,与TST相比,差异显著(P0.05),而其他方法的阳性检出率均低于TST。结果表明,IFN-γ-ELISA方法的检测敏感性较高,在牛结核病污染场净化初期使用,可以尽量避免阳性牛漏检,从而加快牛结核病净化进程。     

上海市某奶牛场牛型结核菌素皮内变态反应疑似结果影响因素分析

为评估奶牛年龄分布、棚舍位置、胎次、泌乳等因素,对牛型结核菌素(PPD)皮内变态反应试验特异性的影响,对上海市某奶牛场2 031头奶牛的PPD皮内变态反应试验初筛结果进行分析和回顾性问卷调查,并利用Epi info~(TM) 7软件,对初筛结果和问卷调查结果进行单因素分析。初筛和问卷调查结果显示:该奶牛场PPD皮内变态反应试验初筛疑似反应率为15.75%,疑似反应主要集中在2岁以下、0～2胎的青年奶牛,其中1.5～2岁青牛奶牛疑似反应率最高;随着年龄、泌乳量和胎次的增加,疑似反应率呈下降趋势。单因素分析结果显示:未配种奶牛(OR=2.02,P0.01),未怀孕奶牛(OR=1.37,P0.01),未泌乳奶牛或干奶牛(OR=3.72,P0.01)是导致奶牛场出现PPD皮内变态反应试验疑似反应的危害性风险因素。结果表明:2岁以下青年奶牛易出现PPD皮内变态反应试验疑似反应,呈现出显著的年龄相关性;是否配种、怀孕以及泌乳量多少均能够影响PPD皮内变态反应试验的特异性。建议奶牛场将犊牛结核病首次检测日龄提前至90日龄以内,并对初筛出现疑似反应的奶牛立即隔离,采用γ-干扰素试验和PPD比较皮内变态反应试验的平行检测策略进行确诊。本研究全面了解了奶牛场初筛疑似反应奶牛的个体特征和风险因素,有助于提高检测方法的敏感性,减少假阴性。     

皮内变态反应试验检测奶牛结核病的假阳性率初步测定及对策

笔者用牛型提纯结核菌素皮内变态反应试验对5425头奶牛进行结核病监测检测,然后间隔42d对前者检测出的阳性牛或可疑牛再次进行结核病检测,比较前后两次的阳性牛或可疑牛符合率.结果显示,前次应用皮内变态反应试验共检测出结核病阳性牛或可疑牛共计142头;再次检测呈阳性或可疑的142头奶牛中,结果仍为阳性牛121头.结果表明,皮内变态反应试验存在一定的假阳性,假阳性率约为14.8%(21/142),建议用皮内变态反试验监测检测奶牛结核病时,对检测出的阳性牛开展1次复核,以提高结核病检测的准确性和降低养殖业主的经济损失.     

应用皮内变态反应和γ-干扰素体外释放试验检测奶牛结核病的比较研究

本研究首先应用皮内变态反应对10200头奶牛进行结核病检测,然后应用γ-干扰素体外释放试验对前者检测出的阳性牛和可疑牛再次进行结核病检测,比较两者的阳性符合率。结果显示,应用皮内变态反应共检测出结核病阳性牛96头、可疑牛4头;γ-干扰素体外释放试验检测皮内变态反应呈阳性的96头奶牛,结果为阳性牛94头、阴性牛2头,而检测皮内变态反应为可疑的4头奶牛,结果全为阳性。结果表明,两种方法的阳性符合率为97．92％（94／96）,虽然皮内变态反应存在一定的假阳性,且费时、费力,但考虑到γ-干扰素试剂盒比较昂贵,建议用皮内变态反应作为初筛试验,γ-干扰素试验用于初筛阳性样品的复核,以提高结核病检测的准确性。     

γ-干扰素ELISA检测方法在牛结核病流行病学调查中的应用

[目的 ]了解宁夏贺兰县奶牛结核病的流行情况,为制定宁夏地区牛结核病净化方案提供参考。[方法 ]2015年采用皮内变态反应和γ-干扰素ELISA检测方法,对宁夏贺兰县2个规模化奶牛场开展牛结核病流行病学调查,共检测奶牛2 230头。[结果 ]在2个规模化奶牛场累计检测出皮内变态反应阳性奶牛79头,皮内变态反应和γ-干扰素ELISA双阳性奶牛72头,双阳性率为3.23%(72/2 230)。[结论 ]γ-干扰素ELISA检测方法准确度较高;在对牛群进行结核病检测时,先以皮内变态反应方法筛测,再结合γ-干扰素ELISA检测结果确诊,更有利于牛结核病的检测与净化。     

两种方法检测云南奶水牛结核病的应用研究

将PPD皮内变态反应法和抗γ-干扰素ELISA法相结合,对云南奶水牛进行牛结核病检测。应用PPD皮内变态反应法检测304头奶水牛,检出32头阳性、25头疑似反应牛,检出阳性和可疑比例高达18．75％,阳性和可疑牛只的分布并无地域和场的趋向性。对PPD皮内变态反应法检出的阳性和疑似反应牛经采集抗凝全血样品54份,应用抗γ-干扰素ELISA法复检,未检检出阳性牛只。结合对受抗γ-干扰素ELISA法检测牛的临床观察,初步认为PPD皮内变态反应法检出阳性和可疑牛系非特异性反应所致。     

二种牛型提纯结核菌素变态反应试验结果比较

为了解不同厂家生产的牛型提纯结核菌素（PPD）对奶牛结核病检疫皮内变态反应试验结果的影响,随机选取二个不同规模奶牛场的100头青年母牛,分别采用国产和荷兰产PPD诊断液2000IU和3000IU的剂量,在每头参试牛的左右侧颈部同时进行皮内变态反应试验,并对判定结果进行比较;同时对所有阳性牛肺门淋巴结样品应用细菌培养的方法进行复核。结果显示：二种PPD诊断液均存在非特异性反应,但国产PPD的非特异性反应显著大于荷兰产PPD（P〈O．01）。     

PPD的批次与注射方式对PPD皮内变态反应检疫的影响

为了探究在牛结核病检疫中出现的使用不同批次的PPD和皮下注射PPD等情况是否会对检疫结果的判定造成干扰。本试验首次通过在豚鼠背部采用皮下注射法和皮内交替注射同一厂家不同批次的结核菌素提纯蛋白衍化物(Purified protein derivative of tuberculin,PPD)的方法,参照《结核菌素提纯蛋白衍化物(PPD)制造及检定规程》于每次注射后24h、48h和72h观察皮试处是否产生迟发型变态反应。试验结果为皮内组(不同批次PPD皮内注射)和皮下组(同一批次PPD皮下注射)经3次注射后的皮试结果与标准参照组的皮试结果无差异,且在每次皮试后的24h、48h和72h皮试处均未出现局部红斑硬结。本试验证明了PPD皮内变态反应检疫结果判定不受PPD皮下接种及PPD的不同批次的干扰。     

牛型结核菌素皮内变态反应试验和γ-干扰素ELISA试验检测奶牛结核病的比较研究

本研究采用PPD皮内变态反应试验和γ-干扰素ELISA试验,对甘肃省3个地区的1585头奶牛进行结核病检测.结果表明,PPD皮内变态反应共检出7份阳性样品;经γ-干扰素ELISA检测2份为阳性、其余5份为假阳性或禽型阳性,假阳性或禽型阳性样品再经细菌分离鉴定表现为阴性;PPD皮内变态反应检出的21份疑似样品再经γ-干扰素ELISA检测,表现为禽型阳性、假阳性或阴性;PPD皮内变态反应阴性样品经γ-干扰素ELISA试验检测,结果为阴性或禽型阳性.在检测奶牛结核病时,PPD皮内变态反应试验特异性较差,γ-干扰素ELISA试验结果与牛结核分枝杆菌细菌分离鉴定结果一致,而且该技术敏感性、特异性和鉴别假阳性均优于PPD皮内变态反应试验.     

一起奶牛结核病检疫阳性的处理

文章旨在通过介绍一起奶牛养殖场在结核病检疫中,用皮内变态反应试验对261头奶牛进行检测,检测出3头阳性牛,并对其进行无害化处理,采取综合防制措施,净化奶牛场结核病,提高养殖效益。     

Comparison between IMMUNODOT tests and the intradermal skin test in atopic dogs

This study evaluated a new perspective in the diagnosis of dermatitis in dogs with signs suggestive of allergic skin disease. The results obtained with CMG IMMUNODOT tests using the technique of allergen-specific strip tests, as employed for human allergy diagnosis, were compared with those obtained by the intradermal skin test (IDST). Forty-eight cases completed the diagnostic evaluation, which included IDST, flea-control program, exclusion of sarcoptes and, for some cases, a 1- to 2-month stabilization period on a restricted protein source diet and testing the serum in the presence of allergen-specific IgE and total IgE. The most common disorders included house and storage dust mites, allergic dermatitis and flea-allergic dermatitis together with atopy. This was confirmed serologically. In the case of positive IDST to pollens, Aspergillus spp. and cat epithelium, CMG IMMUNODOT strip tests were negative. A total of 25% of cases were considered to be primarily associated with food hypersensitivity, but only 4% were confirmed serologically. This study emphasizes the value of CMG IMMUNODOT tests as a support in the diagnosis of dog allergy.     

Diagnosis of flea allergy dermatitis: comparison of intradermal testing with flea allergens and a FcepsilonRI alpha-based IgE assay in response to flea control

The objective of this study was to evaluate the accuracy of in vivo and in vitro tests in the diagnosis of flea allergy dermatitis in comparison with history, clinical signs and response to flea control. Intradermal testing using four different sources of flea allergens and FcepsilonRIalpha-based immunoglobulin (Ig)E assays were performed in 15 flea-allergic dogs, 15 atopic dogs and 15 dogs infested with fleas but showing no clinical signs of skin disease. Sensitivity, specificity, negative predictive value, positive predictive value and accuracy were calculated for all five tests and results varied greatly. Sensitivity, specificity and overall accuracy were 27, 83 and 64%, respectively, for one extract (Isotec), 67, 90 and 82% for another extract (Greer), 93, 90 and 91% for flea saliva, 40, 90 and 73% for the recombinant Cte f 1 both produced by Heska Corp. and 87, 53 and 64% for a FcepsilonRIalpha-based IgE assay. These results indicate that intradermal testing with flea extracts is more accurate in the diagnosis of flea allergy dermatitis than in vitro tests. Moreover, pure flea saliva used as a reagent for intradermal testing provided the best results in terms of sensitivity, specificity and overall accuracy although the Greer extract, a whole body flea extract, also allowed a good correlation between intradermal testing results and clinical approach to flea allergy dermatitis diagnosis.     

Effects of propofol-induced sedation on intradermal test reactions in dogs with atopic dermatitis

We compared the effect of propofol and saline control on intradermal test reactions in dogs with atopic dermatitis undergoing outpatient intradermal testing (IDT). Nineteen dogs were used in this clinical study. Patients were randomly allocated to receive either intravenous (IV) propofol or IV 0.9% saline, and IDT was performed on the right or left (randomized) lateral thorax. One investigator, unaware of the treatments, interpreted all IDT results. Injection sites were analysed using a subjective and objective method. A value of P or= 1+ on all dogs, significantly more positive sites were apparent during propofol sedation than during saline administration. In addition, the greater number of individual dogs experiencing more positive reactions >or= 1+ during propofol sedation was significant. When subjectively analysing reactions >or= 2+, the greater number of positive reactions and the greater number of dogs with more positive reactions observed during propofol treatment was not significantly different from the saline control. When analysed objectively, the greater number of positive reactions observed during propofol sedation was not significant. A greater number of dogs had higher subjective scores and larger objective measurements during propofol sedation compared with saline administration. In summary, propofol sedation was associated with an overall greater number of positive IDT reactions compared with the saline control. Although not always significant, this difference should be considered when choosing propofol for skin testing dogs with atopic dermatitis.     

Evaluation of relationship between plasma fibrinogen concentration and tuberculin testing in red deer

After intradermal injection of bovine purified derivative (PPD), increases in plasma fibrinogen concentration and plasma viscosity developed in red deer (Cervus elaphus) with a history of tuberculosis caused by Mycobacterium bovis. Serum haptoglobin concentrations were also found to increase under similar circumstances. The increases were reproducible and did not appear to be related to mustering, stress, or the handling associated with injection of PPD. A significant (P less than 0.05) direct relationship was found between the increase in plasma fibrinogen concentration and various markers of bovine tuberculosis infection, such as stimulation of lymphocyte transformation in response to bovine PPD and the diameter of intradermal tuberculin skin test reactions. A stronger correlation (P less than 0.01) was found with the volume of intradermal tuberculin skin test reactivity, and the strongest correlation (P less than 0.001) was with the presence of circulating antibovine PPD antibody.     

Evaluation of two cocktails containing ESAT-6, CFP-10 and Rv-3615c in the intradermal test and the interferon-γ assay for diagnosis of bovine tuberculosis

The intradermal tuberculin tests and the interferon-gamma (IFN-γ) assay are the principal tests used worldwide for the ante-mortem diagnosis of bovine tuberculosis. The conventional reagent currently in use in these tests is purified protein derivative (PPD) tuberculin obtained from Mycobacterium bovis culture. The components of PPD are poorly characterized and difficult to standardize. To overcome this issue, antigens specific to the Mycobacterium tuberculosis complex are being studied. Here we have assessed the biological potency of ESAT-6, CFP-10 and Rv-3615c presented as peptide or recombinant protein cocktails in comparison with the standard bovine PPD used routinely in Spanish eradication campaigns. The study was performed in cattle (n=23) from a herd with natural M. bovis infection. Animals were simultaneously injected with PPD and the peptide and protein cocktails. The percentages of cattle reacting positively to single intradermal test were 60.9% (bovine PPD), 47.8% (peptide cocktail) and 60.9% (protein cocktail), with no significant difference between the actual skin fold thickness increases (p>0.05). The IFN-γ assay detected 60.9% of animals when stimulation was performed with bovine PPD, but decreased to 52.2% when stimulation was performed with the peptide cocktail and to 47.8% when stimulation was performed with the protein cocktail. However, no significant differences were found between IFN-γ responder frequencies (p>0.05). These results show a potential use of these defined reagents for in vivo tuberculosis diagnosis.     

Evaluation of an ELISA-PPD for the diagnosis of bovine tuberculosis in field trials in Brazil.

Bovine tuberculosis is a major health problem in Brazil. The intradermal tuberculin test is the standard test for its detection, but it can lack both sensitivity and specificity. The purpose of this study was to evaluate a bovine enzyme-linked immunosorbent assay- (ELISA - PPD) under field conditions in Brazil. A total of 1632 animals from 13 dairy farms were tested with the intradermal tuberculin test (ITT). Two hundred and seven cows gave a positive reaction, which represents 12.7 per cent of the cattle studied. The sensitivity and specificity rates to ITT were 87.7 per cent and 95.2 per cent, respectively. From the 1632 animals 15 per cent of each herd (220 in total) were selected to be tested by the ELISA. Differences between mean optical density (OD) of the control group, ITT -positive and ITT -negative groups were all significant (P<0.01). The sensitivity rates to ELISA - PPD were 86.7 per cent, while specificity was 90.6 per cent. The use of ELISA - PPD is suggested for situations where the investigation of the whole herd is more important than the individual testing of each cow. In addition, the ELISA - PPD can also be helpful when a collective diagnosis is desired to elucidate clinical suspicions of disease, or in the first steps of a control program, for identification of foci.