排除牛结核PPD-ELISA非特异性反应研究
摘要
本试验利用凝胶层析和亲和层析等不同的免疫学方法对PPD抗原进行处理，制备了层析抗原1、层析抗原2和亲和抗原同时通过ELISA方法比较了PPD处理抗原的抗原特异性，并用草分枝杆菌吸收抗原吸收待检血清中的非特异性抗体。结果表明亲和抗原最具特异性，用草分枝杆菌吸收抗原吸收差异显著。
本试验确定了最佳ELISA检测方法并对141头结核变态反应阳性牛和124头变态反应阴性牛进行牛结核ELISA检测，认为由于PPD亲和抗原特异性的增强和排除血清中草分枝杆菌等非特异性的干扰，使试验达到了排除非特异性反应的目的。本试验将为牛结核ELISA诊断提供重要的理论依据。
Obtained gum chromatography antigen 1, gum chromatography antigen 2 and immune-affinity antigen make use of the immunology methods of the gum chromatography and immune-affinity chromatography to handle to PPD antigen and compared the specify of antigens in this experiment, at same time absorbed non-specific antibody with the absorption antigen of M.phlei. It proved that PPD immune-affinity chromatography antigen is the best antigen. It is prominent difference when absorbed the serum with other types specific antigen.
Make sure of the best method of ELISA examination at the same time in this experiments. Examined 141 heads of cow with tubercles allergy positive reaction and 124 heads of cow tubercles allergy negative reaction, This experiments achieved the purpose for eliminiate non-specific responding through eliminated interference of other types specify antibody such as M.phlei and strengthen the antigen specify. This experiment is the importance theories basis for the cow tubercle ELISA examination.
本试验确定了最佳ELISA检测方法并对141头结核变态反应阳性牛和124头变态反应阴性牛进行牛结核ELISA检测，认为由于PPD亲和抗原特异性的增强和排除血清中草分枝杆菌等非特异性的干扰，使试验达到了排除非特异性反应的目的。本试验将为牛结核ELISA诊断提供重要的理论依据。
Obtained gum chromatography antigen 1, gum chromatography antigen 2 and immune-affinity antigen make use of the immunology methods of the gum chromatography and immune-affinity chromatography to handle to PPD antigen and compared the specify of antigens in this experiment, at same time absorbed non-specific antibody with the absorption antigen of M.phlei. It proved that PPD immune-affinity chromatography antigen is the best antigen. It is prominent difference when absorbed the serum with other types specific antigen.
Make sure of the best method of ELISA examination at the same time in this experiments. Examined 141 heads of cow with tubercles allergy positive reaction and 124 heads of cow tubercles allergy negative reaction, This experiments achieved the purpose for eliminiate non-specific responding through eliminated interference of other types specify antibody such as M.phlei and strengthen the antigen specify. This experiment is the importance theories basis for the cow tubercle ELISA examination.
引文
[1]蔡宝祥．家畜传染病(第三版)．中国农业出版社，2000，81-84
[2]张崇贤．什么阻止我们消除结核病?结核与肺部疾病杂志·中文版，1997，1(1)：3-12
[3]何昭阳．牛结核病研究进展．预防兽医学研究进展，2001，3(4)：34-39
[4]戴云海．多重耐药性结核病现状、机制与诊治，国外医学微生物分册，1995，18(5)：32-34
[5]蔡宏．牛结核病流行病学和分子生物学研究进展，中国兽医杂志，1996，22(10)：45-47
[6]张高迪．内蒙古哲里木盟人与牛结核相互传播关系的研究，内蒙古畜牧兽医，1996，2：(3)：47-49
[7]Jorge Hernandez, Mv Z, et al., Effect of Tuberculosis on milk production in dairy cows, JAVMA, 1998,213(6):851-854
[8]Renteria Evangelista TB, Hernandez de Anda J.Tuberculosis in dairy calves:risk of Mycobacterium spp exposure associated with management of colostrums and milk, Prey Vet Med, 1996,27(1):23-27
[9]Brown WH, Hernandez de Anda J.Tuberculosis in adult beef cattle of Mixican origin shipped direct-to-slaughter into Texas, J Am Vet Med Assoc, 1998,212(11):557-559
[10]何昭阳．奶牛结核病的防制．吉林畜牧兽医，1992，3：37-40
[11]何昭阳，胡希荣，等．结核变态阳性牛的非典型分枝杆菌的分离与鉴定．畜牧兽医学报，1994，25(3)：247-251
[12]王振国、罗雁非，等．牛源非典型分枝杆菌的分离与鉴定．吉林农业大学学报，1998，20(2)：73-77
[13]柳春华．应用提纯牛型结核菌素(PPD)和结核菌素(OT)皮内试验对成年奶牛检疫的观察．家畜传染病，1986，26(1)：27-28
[14]Giron D O. Diagnosis of bovine tuberculosis. Noticampo 1997,4(1):22-28
[15]Joardar S N, Sikdar A, et al. Immunodiagnostic approaches to detect bovine tuberculosis. Current Science, 1996,71 (5): 376-378
[16]Machackova M, Lamka J. et al. Bovine tuberculosis in hoofed game animals. Veterinarstvi 2000 50(9):349-55
[17]Ryan T J, Buddle B M, et al. An evaluation of the gamma interferon test for detecting bovine tuberculosis in cattle 8 to 28 days after tuberculin skin testing. Research in Veterinary Science. 2000,69(1):57-61
[18]Scacchia M, Lelli R, et al. Use of innovative methods in the eradication of bovine tuberculosis. Journal of Veterinary Medicine.2000,47(5): 321-27
[19]Pollock J M, Girvin R M, et al. Assessment of defined antigens for the diagnosis of bovine tuberculosis in skin test-reactor cattle. Veterinary Record 2000,146(23):659-665
[20]Briones V, Juan L DE, et al. Bovine tuberculosis and the endanged Iberian Iynx. Emerging Infectious Diseases, 2000,6(2): 189-191
[21]Abernetihy D A, Pfeiffer D U J, et al. Case control study examining the role of livestock markets in the transmission of bovine tuberculosis. Society for Veterinary Epidemiology and Preventive Medicine 1999,10(1):108-114
[22]Scacchia M, Lelli R. wt al. Evaluation of new and attendant diagnostic tests for surveillance in the final part of eradication program for bovine tuberculosis. Veterinaria ltaliana, 1996,32(22):51-56
[23]Nelson A M. The cost of disease eradication: smallpox and bovine tuberculosis. Annals of the New York Academy of science, 1999,894(1):83-91
[24]Emmerzaal A, Deleu S, et al. Cattle tuberculosis [caused by Mycobacterium bovine] is difficult to detect and to treat. Veehouder en Dierenarts, 2000,14(2):4-7
[25]王发锁．结核病的实验诊断进展，海军军事医学，1995，16(4)：46-48
[26]O.R.Gonzalez Llamazares, C.B.Gutierrez Martin, et al,. Field evaluation of the single intradermal cervical tuberculin test and the interferon-γ assay for detection and eradication of bovine tuberculosis in Spain, Veterinary Microbiology, 1999,70(1):55-66
[27]Milian-Suazo F, Salman M, et al,. Molecular epidemiologic analysis of Mycobacterium bovis isolates from Mexico, American Journal of Vterinary Research, 2000,61(1):90-95
[28]吴雪琼．分枝杆菌DNA探针研究及应用概况，微生物学通报，1992，19(4)：230-233
[29]庄玉辉，吴雪琼，等．聚合酶链式反应检测结核杆菌的研究，微生物学通报，1992,32(5)：364-369
[30]刘雪平．气相色谱检测脑脊液中结核分枝杆菌硬脂酸对结核性脑膜炎诊断的研究，中华结核和呼吸杂志，1994,17(1)：32-35
[31]Thon C O, Hall M B, et al. Proc Anim Health assoc usa 1983,87(1):603-610
[32]逄魁春．牛结核ELISA及其抗原的比较研究，预防兽医学硕士研究生论文集(2)
[33]柴方静．BA-ELISA检测血．清抗体的研究，中国畜禽传染病，1991，5(1)：31-34
[34]赵新泰．应用Dot-ELISA检测结核病牛血清抗体效价，中国畜禽传染病，1988，3(1)：33-34
[35]胡忠义．用生物素标记结核菌单克隆抗体检测脑脊液结核菌抗原的研究，微生物学通报 1990，17，(4)，226-228
[36]王彩平．BA-Dot-ELISA检测结核病牛血清抗体的研究，吉林农业大学学报，1992，14(3)：75-78
[37]宋立彪，韩美华．ABC-ELISA法检测脑脊液抗PPD-IgG及其临床意义，中华结核和呼吸杂志，1994，17(2)：82-83
[38]邹伟文．酶联免疫吸附试验检测结核抗体实验条件的比较，江西医学院学报，1995，35(3)30-34
[39]Julian E. Matas A, et al,. Evaluation of a new serodiagnostic tuberculosis test based on immuneoglobulin A detection against Kp-90 antigen, International Journal of tuberculosis and Lung Disease, 2000,4(11):1082-1085
[40]Alifano M, De-Pascalis R, et al,. Detection of IgG and IgA against the mycobacterial antigen A60 in patients with extrapulmonary tuberculosis, Thorax, 1998, 53(5):377-380
[41]胡姣荣．脑脊液铁蛋白检测方法的改进(微量酶联免疫吸附法)，湖北医科大学学报，1994，15(3)：276-279
[42]侯伟．两种糖脂抗原在结核病血清诊断中的应用，中国医学科学院学报，1994，16(5)374-377
[43]张景道．检测血清中BCG-A60抗体对结核病诊断的临床意义，中华结核和呼吸杂志，1994，17(3)：175-178
[44]林松柏．脂阿拉伯甘露糖(LAM)用于血清学诊断活动性结核病的评价，中国防痨杂志，1995、17(4)：168-169
[45]郝世家．结明(MycoDotTM)快捷试验对菌阴肺结核诊断的价值，中国防痨杂志，1997，19(4)：191-192
[46]侯伟．Tb-NT-P-BSA抗原在结核病血清诊断的初步评价，中国人兽共患病杂志，1999，15，(1)：43-44
[47]Chan ED, Reves R, et al,. Diagnosis of tuberculosis by a visually detectable immunoassay for lipoarabinomannan, 2000, 161(5):1713-1719
[48]Tessema TA, Hamasur B, et al,. Diagnostic evaluation of urinary lipoarabinomannan at an Ethiopian tuberculosis center, Scandinavian Journal of Infectious diseases, 2001,33(4):279-284
[49]熊礼宽．血清38KDIgG抗体和分枝杆菌IgG抗体检测对肺结核诊断价值的探讨，深圳医
学，1996，9(5) :7-8
[50] Sakai .J, Matsuzawa S, et al,. New diagnostic approach for ocular tuberculosis by ELISA using the cord factor as antigen, British Journal of Ophthalmology, 2001,85(2) : 130-133
[51] Fifis T, Costopoulos C, et al,. Purification and characterization of a major antigen from Mycobacterium bovine culture filtrate, Infect Immum, 1990,59(5) : 800-807
[52] Lyashchenko K, Pollock J, et al,. Diversity of antigen recognition by serum antibodies in experimental bovine tuberculosis, 1998,66(11) :5344-5349
[53] Pollock J, Andersen P,. The potential of the ESAT-6 antigen secreted by virulent mycobacteria for specific diagnosis of tuberculosis, Journal of Infectious Diseases, 1997,175(5) :1251-1254
[54] Lightbody K, Skuce R, et al,. Mycobacterial antigen-specific antibody responses in bovine tuberculosis: an ELISA with potential to confirm disease status, Veterinary Record, 1998, 1 42(12) :295-300
[55] Rhodes SG, Gavier-Widen D, et al,. Antigen specificity in experimental bovine tuberculosis, 2000,68(5) :2573-2578
[2]张崇贤．什么阻止我们消除结核病?结核与肺部疾病杂志·中文版，1997，1(1)：3-12
[3]何昭阳．牛结核病研究进展．预防兽医学研究进展，2001，3(4)：34-39
[4]戴云海．多重耐药性结核病现状、机制与诊治，国外医学微生物分册，1995，18(5)：32-34
[5]蔡宏．牛结核病流行病学和分子生物学研究进展，中国兽医杂志，1996，22(10)：45-47
[6]张高迪．内蒙古哲里木盟人与牛结核相互传播关系的研究，内蒙古畜牧兽医，1996，2：(3)：47-49
[7]Jorge Hernandez, Mv Z, et al., Effect of Tuberculosis on milk production in dairy cows, JAVMA, 1998,213(6):851-854
[8]Renteria Evangelista TB, Hernandez de Anda J.Tuberculosis in dairy calves:risk of Mycobacterium spp exposure associated with management of colostrums and milk, Prey Vet Med, 1996,27(1):23-27
[9]Brown WH, Hernandez de Anda J.Tuberculosis in adult beef cattle of Mixican origin shipped direct-to-slaughter into Texas, J Am Vet Med Assoc, 1998,212(11):557-559
[10]何昭阳．奶牛结核病的防制．吉林畜牧兽医，1992，3：37-40
[11]何昭阳，胡希荣，等．结核变态阳性牛的非典型分枝杆菌的分离与鉴定．畜牧兽医学报，1994，25(3)：247-251
[12]王振国、罗雁非，等．牛源非典型分枝杆菌的分离与鉴定．吉林农业大学学报，1998，20(2)：73-77
[13]柳春华．应用提纯牛型结核菌素(PPD)和结核菌素(OT)皮内试验对成年奶牛检疫的观察．家畜传染病，1986，26(1)：27-28
[14]Giron D O. Diagnosis of bovine tuberculosis. Noticampo 1997,4(1):22-28
[15]Joardar S N, Sikdar A, et al. Immunodiagnostic approaches to detect bovine tuberculosis. Current Science, 1996,71 (5): 376-378
[16]Machackova M, Lamka J. et al. Bovine tuberculosis in hoofed game animals. Veterinarstvi 2000 50(9):349-55
[17]Ryan T J, Buddle B M, et al. An evaluation of the gamma interferon test for detecting bovine tuberculosis in cattle 8 to 28 days after tuberculin skin testing. Research in Veterinary Science. 2000,69(1):57-61
[18]Scacchia M, Lelli R, et al. Use of innovative methods in the eradication of bovine tuberculosis. Journal of Veterinary Medicine.2000,47(5): 321-27
[19]Pollock J M, Girvin R M, et al. Assessment of defined antigens for the diagnosis of bovine tuberculosis in skin test-reactor cattle. Veterinary Record 2000,146(23):659-665
[20]Briones V, Juan L DE, et al. Bovine tuberculosis and the endanged Iberian Iynx. Emerging Infectious Diseases, 2000,6(2): 189-191
[21]Abernetihy D A, Pfeiffer D U J, et al. Case control study examining the role of livestock markets in the transmission of bovine tuberculosis. Society for Veterinary Epidemiology and Preventive Medicine 1999,10(1):108-114
[22]Scacchia M, Lelli R. wt al. Evaluation of new and attendant diagnostic tests for surveillance in the final part of eradication program for bovine tuberculosis. Veterinaria ltaliana, 1996,32(22):51-56
[23]Nelson A M. The cost of disease eradication: smallpox and bovine tuberculosis. Annals of the New York Academy of science, 1999,894(1):83-91
[24]Emmerzaal A, Deleu S, et al. Cattle tuberculosis [caused by Mycobacterium bovine] is difficult to detect and to treat. Veehouder en Dierenarts, 2000,14(2):4-7
[25]王发锁．结核病的实验诊断进展，海军军事医学，1995，16(4)：46-48
[26]O.R.Gonzalez Llamazares, C.B.Gutierrez Martin, et al,. Field evaluation of the single intradermal cervical tuberculin test and the interferon-γ assay for detection and eradication of bovine tuberculosis in Spain, Veterinary Microbiology, 1999,70(1):55-66
[27]Milian-Suazo F, Salman M, et al,. Molecular epidemiologic analysis of Mycobacterium bovis isolates from Mexico, American Journal of Vterinary Research, 2000,61(1):90-95
[28]吴雪琼．分枝杆菌DNA探针研究及应用概况，微生物学通报，1992，19(4)：230-233
[29]庄玉辉，吴雪琼，等．聚合酶链式反应检测结核杆菌的研究，微生物学通报，1992,32(5)：364-369
[30]刘雪平．气相色谱检测脑脊液中结核分枝杆菌硬脂酸对结核性脑膜炎诊断的研究，中华结核和呼吸杂志，1994,17(1)：32-35
[31]Thon C O, Hall M B, et al. Proc Anim Health assoc usa 1983,87(1):603-610
[32]逄魁春．牛结核ELISA及其抗原的比较研究，预防兽医学硕士研究生论文集(2)
[33]柴方静．BA-ELISA检测血．清抗体的研究，中国畜禽传染病，1991，5(1)：31-34
[34]赵新泰．应用Dot-ELISA检测结核病牛血清抗体效价，中国畜禽传染病，1988，3(1)：33-34
[35]胡忠义．用生物素标记结核菌单克隆抗体检测脑脊液结核菌抗原的研究，微生物学通报 1990，17，(4)，226-228
[36]王彩平．BA-Dot-ELISA检测结核病牛血清抗体的研究，吉林农业大学学报，1992，14(3)：75-78
[37]宋立彪，韩美华．ABC-ELISA法检测脑脊液抗PPD-IgG及其临床意义，中华结核和呼吸杂志，1994，17(2)：82-83
[38]邹伟文．酶联免疫吸附试验检测结核抗体实验条件的比较，江西医学院学报，1995，35(3)30-34
[39]Julian E. Matas A, et al,. Evaluation of a new serodiagnostic tuberculosis test based on immuneoglobulin A detection against Kp-90 antigen, International Journal of tuberculosis and Lung Disease, 2000,4(11):1082-1085
[40]Alifano M, De-Pascalis R, et al,. Detection of IgG and IgA against the mycobacterial antigen A60 in patients with extrapulmonary tuberculosis, Thorax, 1998, 53(5):377-380
[41]胡姣荣．脑脊液铁蛋白检测方法的改进(微量酶联免疫吸附法)，湖北医科大学学报，1994，15(3)：276-279
[42]侯伟．两种糖脂抗原在结核病血清诊断中的应用，中国医学科学院学报，1994，16(5)374-377
[43]张景道．检测血清中BCG-A60抗体对结核病诊断的临床意义，中华结核和呼吸杂志，1994，17(3)：175-178
[44]林松柏．脂阿拉伯甘露糖(LAM)用于血清学诊断活动性结核病的评价，中国防痨杂志，1995、17(4)：168-169
[45]郝世家．结明(MycoDotTM)快捷试验对菌阴肺结核诊断的价值，中国防痨杂志，1997，19(4)：191-192
[46]侯伟．Tb-NT-P-BSA抗原在结核病血清诊断的初步评价，中国人兽共患病杂志，1999，15，(1)：43-44
[47]Chan ED, Reves R, et al,. Diagnosis of tuberculosis by a visually detectable immunoassay for lipoarabinomannan, 2000, 161(5):1713-1719
[48]Tessema TA, Hamasur B, et al,. Diagnostic evaluation of urinary lipoarabinomannan at an Ethiopian tuberculosis center, Scandinavian Journal of Infectious diseases, 2001,33(4):279-284
[49]熊礼宽．血清38KDIgG抗体和分枝杆菌IgG抗体检测对肺结核诊断价值的探讨，深圳医
学，1996，9(5) :7-8
[50] Sakai .J, Matsuzawa S, et al,. New diagnostic approach for ocular tuberculosis by ELISA using the cord factor as antigen, British Journal of Ophthalmology, 2001,85(2) : 130-133
[51] Fifis T, Costopoulos C, et al,. Purification and characterization of a major antigen from Mycobacterium bovine culture filtrate, Infect Immum, 1990,59(5) : 800-807
[52] Lyashchenko K, Pollock J, et al,. Diversity of antigen recognition by serum antibodies in experimental bovine tuberculosis, 1998,66(11) :5344-5349
[53] Pollock J, Andersen P,. The potential of the ESAT-6 antigen secreted by virulent mycobacteria for specific diagnosis of tuberculosis, Journal of Infectious Diseases, 1997,175(5) :1251-1254
[54] Lightbody K, Skuce R, et al,. Mycobacterial antigen-specific antibody responses in bovine tuberculosis: an ELISA with potential to confirm disease status, Veterinary Record, 1998, 1 42(12) :295-300
[55] Rhodes SG, Gavier-Widen D, et al,. Antigen specificity in experimental bovine tuberculosis, 2000,68(5) :2573-2578