臺灣博碩士論文加值系統

新型30分鐘西方點墨法
西方點墨法 (Western blot)是一重要且廣泛運用於特定蛋白質分析的技術。通常蛋白質需在SDS-PAGE凝膠上經電泳方式分離後，再轉漬於硝化纖維膜 (nitrocellulose membrane)或是polyvinylidene difluoride (PVDF)進一步作西方點墨法分析。使用傳統的西方點墨法分析蛋白質通常室溫下需3~4小時操作時間，本研究利用奈米金膠(colloidal gold)自我組成特性(auto-assembly)與抗體形成免疫金膠(immunalgold)，並結合創新的層析技術以達到發展一快速新型西方點墨法的目的。在此利用生乳與人類血漿作為實驗模式，用快速新型西方點墨法與金膠標定��-乳球蛋白(��-lactoglobulin ; ��-LG)的單株抗體可在30分鐘內分析出生乳中的��-乳球蛋白，在此方法中使用另一金膠標定載脂蛋白AІ (apolipoprotein-AІ ; Apo-AІ)與白蛋白(Albumin)的多株抗體的也可快速完成分析人類血漿中的脂蛋白AІ與白蛋白，這創新的”西方金墨法”不但可達到與傳統西方點墨法相同的特異性，更可減少實驗操做時間和節省實驗所需試劑與抗體。

發展幽門螺旋桿菌單株抗體之快速檢測試劑
幽門螺旋桿菌(Helicobacter pylori)會造成胃炎和消化性潰瘍，近年來有研究指出幽門螺旋桿菌也可能是造成胃癌的一個致病因素。檢測幽門螺旋桿菌感染的方法很多，最常用的有五種：細菌培養、組織染色、尿素酶試驗、呼氣試驗及血清學檢查。前三種皆需做胃鏡檢查，屬侵入性檢測方法，後兩種屬非侵入性檢查。運用 (enzyme immunoassay) 直接檢測糞便中幽門螺旋桿菌抗原是另一種新發展的非侵入性檢測法。這種檢測法通常使用辨識幽門螺旋桿菌的兔子多株抗體。
　　本研究目的為發展單株抗體免疫金膠，並結合快速免疫層析法檢測幽門螺旋桿菌抗原，以改進使用多株抗體時的缺點，包含較低的特異性及不穩定的再現性。因此，以幽門螺旋桿菌為抗原給予BALB/c小鼠，在酵素免疫分析法單株抗體篩選過程中以大腸桿菌(E. coli)、耶爾辛氏腸炎桿菌(Y. enterocolitica)、及腸炎弧菌(V. parahaemolyticus)作為陰性控制，而製備出13株不同的幽門螺旋桿菌單株抗體，並以微陣列機(microarray)來尋找最適合的單株抗體配對，最後以單株抗體12F4作為捕捉抗體，而單株抗體11E3作為免疫金膠抗體是最佳組合，以幽門螺旋桿菌抗原為檢體時，可檢測的濃度為10ng/�愮。

第一章 新型30分鐘西方點墨法
目 錄
頁次
目錄----------------------------------------------------------------------------------i
圖目錄-----------------------------------------------------------------------------iii
英文摘要---------------------------------------------------------------------------1
中文摘要---------------------------------------------------------------------------2
第一節 前言---------------------------------------------------------------------3
1.1 西方點墨概論----------------------------------------------------------------3
1.2 奈米科技----------------------------------------------------------------------4
1.3 膠體金-------------------------------------------------------------------------5
1.4 免疫金膠----------------------------------------------------------------------5
1.5 免疫層析----------------------------------------------------------------------6
1.6 本研究之目的----------------------------------------------------------------7

第二節 材料與方法------------------------------------------------------------8
2.1 抗源製備----------------------------------------------------------------------8
2.2 抗體製備-----------------------------------------------------------------------8
2.3 硝化纖維膜-------------------------------------------------------------------8
2.4 操作架設計與製作----------------------------------------------------------8
2.5 SDS-PAGE-------------------------------------------------------------------9
2.6 西方點墨 (Western blot) ---------------------------------------------------9
2.7 新型西方點墨法-----------------------------------------------------------10
2.8 Amido Black 染色---------------------------------------------------------10
2.9 金膠之製作-----------------------------------------------------------------10
2.10 免疫金膠製備--------------------------------------------------------------11

第三節 結果與討論-----------------------------------------------------------11
3.1層析測試---------------------------------------------------------------------11
3.2 8�慆硝化纖維膜測試------------------------------------------------------12
3.3傳統與新型西方點墨法特異性比較------------------------------------12
3.4傳統與新型西方點墨法靈敏度比較------------------------------------13
3.5人類血漿模式---------------------------------------------------------------14

第四節 結論--------------------------------------------------------------------15

第五節 參考文獻---------------------------------------------------------------16


圖 目 錄
頁次
圖1：金膠奈米微粒於溶液中之假說圖-----------------------------------21
圖2：金膠奈米微粒與抗體之間作用力假說圖--------------------------22
圖3：快速新型西方點墨法操作架-----------------------------------------23
圖4：新型西方點墨法操作示意圖-----------------------------------------24
圖5：0.2 �慆硝化纖維膜與8 �慆硝化纖維膜層析測試---------------25
圖6：生乳轉漬至8�慆硝化纖維膜測試結果----------------------------26
圖7：傳統西方點墨法與新型西方點墨法特異性比較結果-----------27
圖8：傳統西方點墨法與新型西方點墨法靈敏度比較結果-----------28
圖9：0.2�慆硝化纖維膜與8�慆硝化纖維膜電顯圖---------------------29
圖10：人類血漿經新型西方點墨法分析結果----------------------------30
圖11：傳統西方點墨法與”西方金墨法”操作時間比較流程圖-------31


第二章 幽門螺旋桿菌單株抗體及其免疫金膠製備
目錄----------------------------------------------------------------------------------i
圖目錄-----------------------------------------------------------------------------iv
表目錄-----------------------------------------------------------------------------iv
附錄---------------------------------------------------------------------------------v
英文摘要---------------------------------------------------------------------------1
中文摘要---------------------------------------------------------------------------2
第一節 前言---------------------------------------------------------------------3
1.1 幽門螺旋桿菌概論----------------------------------------------------------3
1.2 幽門螺旋桿菌之感染-------------------------------------------------------3
1.3 幽門螺旋桿菌相關疾病----------------------------------------------------4
　 1.3.1 胃炎---------------------------------------------------------------------4
1.3.2 消化性潰瘍------------------------------------------------------------5
1.3.3 胃癌---------------------------------------------------------------------6
1.3.4 胃黏膜組織相關淋巴瘤---------------------------------------------7
1.4 幽門螺旋桿菌的清除與治療----------------------------------------------7
1.5 幽門螺旋桿菌的檢測-------------------------------------------------------8
1.5.1幽門螺旋桿菌侵入性檢測方法-------------------------------------9
1.5.2幽門螺旋桿菌非侵入性檢測方法--------------------------------10
1.6 本研究之目的---------------------------------------------------------------11
第二節 材料與方法-----------------------------------------------------------12
2.1 抗源的製備-----------------------------------------------------------------12
2.2 單株抗體之製作-----------------------------------------------------------13
2.2.1 免疫反應--------------------------------------------------------------15
2.2.2 骨髓癌細胞-----------------------------------------------------------15
2.2.3 細胞融合--------------------------------------------------------------16
2.2.4 細胞增殖與取樣篩檢-----------------------------------------------16
2.2.5 融合細胞之單株化--------------------------------------------------17
2.2.6 單株抗體之生產-----------------------------------------------------17
2.2.7 培養液及試劑--------------------------------------------------------18
2.2.8 腹水抗體製備--------------------------------------------------------19
2.3 幽門螺旋桿菌抗體的測定-----------------------------------------------19
2.3.1 ELISA------------------------------------------------------------------19
2.3.2 SDS-PAGE------------------------------------------------------------20
2.3.3 Western blot-----------------------------------------------------------22
2.3.4 抗體isotype-----------------------------------------------------------22
2.4 應用幽門螺旋桿菌單株抗體於免疫層析-----------------------------23
2.4.1 製備免疫金膠--------------------------------------------------------23
2.4.2 製備免疫層析法所使用之檢測片--------------------------------24
2.4.3 幽門螺旋桿菌抗原濃度檢測--------------------------------------24
2.4.4 臨床糞便檢體檢測--------------------------------------------------24
2.5 微陣列系統-----------------------------------------------------------------25
第三節 結果與討論-----------------------------------------------------------25
3.1 單株抗體之製作-----------------------------------------------------------25
3.2 微陣列系統-----------------------------------------------------------------28
3.3 應用幽門螺旋桿菌單株抗體於免疫層析-----------------------------30
3.3.1 免疫層析法原理---------------------------------------------------30
3.3.2 金膠之製造及特性分析------------------------------------------30
3.3.3 免疫層析法之應用------------------------------------------------31
3.3.4幽門螺旋桿菌抗原濃度檢測-------------------------------------31
3.3.5 臨床檢體之檢測---------------------------------------------------32
第四節 結論--------------------------------------------------------------------33
第五節 參考文獻---------------------------------------------------------------34


圖 目 錄
頁次
圖1：微陣列打點機(microarray)系統--------------------------------------41
圖2：單株抗體製作流程-----------------------------------------------------42
圖3：實驗接種小鼠血清力價之測試--------------------------------------43
圖4：融合成功細胞產生的抗體對幽們螺旋桿菌抗原反應篩選-----44
圖5：幽門螺旋桿菌和大腸桿菌的15% SDS-PAGE與幽門螺旋桿菌
單株抗體西方點墨法結果--------------------------------------------45
圖6：幽門螺旋桿菌和大腸桿菌的15% SDS-PAGE與幽門螺旋桿菌
單株抗體西方點墨法結果--------------------------------------------46
圖7：. 腹水抗體力價之測試-------------------------------------------------47
圖8：應用微陣列系統偵測幽門螺旋桿菌單株抗體配對--------------48
圖9：免疫層析法之應用-----------------------------------------------------49
圖10：單株抗體可檢測幽門螺旋桿菌的抗原濃度----------------------50
圖11：抗原與免疫金膠抗體結合示意圖----------------------------------51

表 目 錄
頁次
表1：幽門螺旋桿菌單株抗體之特異性-----------------------------------52
表2：幽門螺旋桿菌單株抗體isotype--------------------------------------53
表3：用免疫金膠檢測臨床糞便檢體結果--------------------------------54





附 錄
頁次
附錄1：實驗設計流程圖-----------------------------------------------------12
附錄2：單株抗體製作流程圖-----------------------------------------------14
附錄3：抗體-抗原-抗體三明治結構示意圖-----------------------------29

新型30分鐘西方點墨法
Adjei, A.A., Armah, H., Duah, O.A., Adiku, T. and Hesse, I.F. (2003) Evaluation of a rapid serological chromatographic immunoassay for the diagnosis of pulmonary tuberculosis in Accra, Ghana. Jpn J Infect Dis 56, 161-4.
Bergendahl, V., Glaser, B.T. and Burgess, R.R. (2003) A fast Western blot procedure improved for quantitative analysis by direct fluorescence labeling of primary antibodies. J Immunol Methods 277, 117-25.
Biggs, S. (1993) The role of colloidal stability in the formation of gold sols. J. Colloid Interface Sci. 160, 511-513.
Brainina, K., Kozitsina, A. and Beikin, J. (2003) Electrochemical immunosensor for Forest-Spring encephalitis based on protein A labeled with colloidal gold. Anal Bioanal Chem 376, 481-5.
Chen W.T., H.M.T., Liu H.C., Li C.W., Mao S.J. (2004) Distinction between dry and raw milk using monoclonal antibodies prepared against dry milk proteins. Journal of Dairy Science. In press.
De Roe, C., Courtoy, P.J. and Baudhuin, P. (1987) A model of protein-colloidal gold interactions. J Histochem Cytochem 35, 1191-8.
Dennis-Sykes, C.A., Miller, W.J. and McAleer, W.J. (1985) A quantitative Western Blot method for protein measurement. J Biol Stand 13, 309-14.
Dubitsky, A., DeCollibus, D. and Ortolano, G.A. (2002) Sensitive fluorescent detection of protein on nylon membranes. J Biochem Biophys Methods 51, 47-56.
Goodman, S.L., Hodges, G.M., Trejdosiewicz, L.K. and Livingston, D.C. (1979) Colloidal gold probes--a further evaluation. Scan Electron Microsc, 619-28.
Inatomi, K. (2003) Application of Amido black staining to enumerating bacteria grown on membrane filters. J Microbiol Methods 55, 393-7.
Johne, B., Hansen, K., Mork, E. and Holtlund, J. (1995) Colloidal gold conjugated monoclonal antibodies, studied in the BIAcore biosensor and in the Nycocard immunoassay format. J Immunol Methods 183, 167-74.
Knecht, D.A. and Dimond, R.L. (1984) Visualization of antigenic proteins on Western blots. Anal Biochem 136, 180-4.
Kurien, B.T. and Scofield, R.H. (2003) Protein blotting: a review. J Immunol Methods 274, 1-15.
Leung, W., Chan, P., Bosgoed, F., Lehmann, K., Renneberg, I., Lehmann, M. and Renneberg, R. (2003) One-step quantitative cortisol dipstick with proportional reading. J Immunol Methods 281, 109-18.
Lindmark, R., Thoren-Tolling, K. and Sjoquist, J. (1983) Binding of immunoglobulins to protein A and immunoglobulin levels in mammalian sera. J Immunol Methods 62, 1-13.
M.A., H. (1989) Colloidal Gold : Principles, Methods, and Applications., 1-17.
Magnusson M.H., D.K., Malm J.O., Bovin J.O. and Samuelson L. (1999) Gold nanoparticles: production, reshaping, and thermal charging. J. Nanoparticle Res. 1, 243-251.
Manaresi, E., Pasini, P., Gallinella, G., Gentilomi, G., Venturoli, S., Roda, A., Zerbini, M. and Musiani, M. (1999) Chemiluminescence Western blot assay for the detection of immunity against parvovirus B19 VP1 and VP2 linear epitopes using a videocamera based luminograph. J Virol Methods 81, 91-9.
Moeremans, M., Daneels, G. and De Mey, J. (1985) Sensitive colloidal metal (gold or silver) staining of protein blots on nitrocellulose membranes. Anal Biochem 145, 315-21.
Morris, R.E. and Saelinger, C.B. (1982) Combined use of immunoferritin and immunocolloidal gold for the simultaneous demonstration of two antigens by immuno-electron microscopy. Stain Technol 57, 225-37.
Oliver, C. (1999a) Conjugation of colloidal gold to proteins. Methods Mol Biol 115, 331-4.
Oliver, C. (1999b) Preparation of colloidal gold. Methods Mol Biol 115, 327-30.
Osikowicz, G., Beggs, M., Brookhart, P., Caplan, D., Ching, S., Eck, P., Gordon, J., Richerson, R., Sampedro, S., Stimpson, D. and et al. (1990) One-step chromatographic immunoassay for qualitative determination of choriogonadotropin in urine. Clin Chem 36, 1586.
Otto, J.J. (1993) Immunoblotting. Methods Cell Biol 37, 105-17.
Rembaum, A. and Dreyer, W.J. (1980) Immunomicrospheres: reagents for cell labeling and separation. Science 208, 364-8.
Robenek, H., Rassat, J., Hesz, A. and Grunwald, J. (1982) A correlative study on the topographical distribution of the receptors for low density lipoprotein (LDL) conjugated to colloid gold in cultured human skin fibroblasts employing thin section, freeze-fracture, deep-etching, and surface replication techniques. Eur J Cell Biol 27, 242-50.
S.C., L. (1998) Biotechnology for nanotechnology. Trends in biotechnology 16, 239-240.
Schneider, B.H., Dickinson, E.L., Vach, M.D., Hoijer, J.V. and Howard, L.V. (2000) Optical chip immunoassay for hCG in human whole blood. Biosens Bioelectron 15, 597-604.
Song, W. and Dou, C. (2003) One-step immunoassay for acetaminophen and salicylate in serum, plasma, and whole blood. J Anal Toxicol 27, 366-71.
Spielberg, F., Kabeya, C.M., Ryder, R.W., Kifuani, N.K., Harris, J., Bender, T.R., Heyward, W.L. and Quinn, T.C. (1989) Field testing and comparative evaluation of rapid, visually read screening assays for antibody to human immunodeficiency virus. Lancet 1, 580-4.
Sundaram, P. (1996) A simplified 'coupled' probing approach for the detection of proteins on dot and western blots. J Immunol Methods 195, 149-52.
Too, C.K., Murphy, P.R. and Croll, R.P. (1994) Western blotting of formaldehyde-fixed neuropeptides as small as 400 daltons on gelatin-coated nitrocellulose paper. Anal Biochem 219, 341-8.
Towbin, H., Staehelin, T. and Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 76, 4350-4.
Valkirs, G.E. and Barton, R. (1985) ImmunoConcentration--a new format for solid-phase immunoassays. Clin Chem 31, 1427-31.
Warchol, J.B. and Brelinska, R. (1980) Some aspects of labelling of proteins with colloidal gold. Acta Med Pol 21, 413-4.
Warchol, J.B., Brelinska, R. and Herbert, D.C. (1982) Analysis of colloidal gold methods for labelling proteins. Histochemistry 76, 567-75.
Wilchek, M. and Bayer, E.A. (1988) The avidin-biotin complex in bioanalytical applications. Anal Biochem 171, 1-32.
Yakunin, A.F. and Hallenbeck, P.C. (1998) A luminol/iodophenol chemiluminescent detection system for western immunoblots. Anal Biochem 258, 146-9.
Yang, S.J.a.S., J. T. Mao. (1999) A simple HPLC Purification Procedure for Porcine Plasma Haptoglobin. J. Chromatagr. B. 731, 395-402.
柯英潔，奈米科技於醫學檢驗之應用：發展快速單一步驟免疫層析法檢測蝦白斑病毒之研究，國立交通大學碩士論文 (2002)。
吳怡荏，發展快速單一步驟免疫層析法檢測血液中低密度脂蛋白濃度之研究，國立交通大學碩士論文 (2003)。

發展幽門螺旋桿菌單株抗體之快速檢測試劑
Alemohammad, M.M., Foley, T.J. and Cohen, H. (1993) Detection of immunoglobulin G antibodies to Helicobacter pylori in urine by an enzyme immunoassay method. J Clin Microbiol 31, 2174-7.
Austin, J.W., Doig, P., Stewart, M. and Trust, T.J. (1991) Macromolecular structure and aggregation states of Helicobacter pylori urease. J Bacteriol 173, 5663-7.
Bhaskar, S., Singh, S. and Sharma, M. (1996) A single-step immunochromatographic test for the detection of Entamoeba histolytica antigen in stool samples. J Immunol Methods 196, 193-8.
Bodger, K., Wyatt, J.I. and Heatley, R.V. (1997) Gastric mucosal secretion of interleukin-10: relations to histopathology, Helicobacter pylori status, and tumour necrosis factor-alpha secretion. Gut 40, 739-44.
Caletti, G., Togliani, T., Fusaroli, P., Sabattini, E., Khodadadian, E., Gamberi, B., Gobbi, M. and Pileri, S. (2003) Consecutive regression of concurrent laryngeal and gastric MALT lymphoma after anti-Helicobacter pylori therapy. Gastroenterology 124, 537-43.
Chu, C., Yu, Y.J., Kong, M.S. and Ou, J.T. (2003) Rate of Helicobacter pylori infection in children and clonality of Taiwan strains. Microbiol Immunol 47, 813-21.
Engvall, E. and Perlman, P. (1971) Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobulin G. Immunochemistry 8, 871-4.
Forne, M., Dominguez, J., Fernandez-Banares, F., Lite, J., Esteve, M., Gali, N., Espinos, J.C., Quintana, S. and Viver, J.M. (2000) Accuracy of an enzyme immunoassay for the detection of Helicobacter pylori in stool specimens in the diagnosis of infection and posttreatment check-up. Am J Gastroenterol 95, 2200-5.
Fukuda, S., Tanaka, M., Soma, Y., Shimoyama, T., Mikami, T., Crabtree, J.E., Saito, H., Munakata, A. and Yoshida, Y. (2000) Histological analysis of gastritis and Helicobacter pylori infection in patients with early gastric cancer: a case-control study. J Gastroenterol Hepatol 15, 1370-6.
Ge, H. (2000) UPA, a universal protein array system for quantitative detection of protein-protein, protein-DNA, protein-RNA and protein-ligand interactions. Nucleic Acids Res 28, e3.
Goh, K.L., Parasakthi, N., Peh, S.C. and Ong, K.K. (1995) 14C-urea breath test: a useful non-invasive test in the diagnosis of Helicobacter pylori infection. Med J Malaysia 50, 208-11.
Goodman, S.L., Hodges, G.M., Trejdosiewicz, L.K. and Livingston, D.C. (1979) Colloidal gold probes--a further evaluation. Scan Electron Microsc, 619-28.
Gotz, J.M., Vergouwe, Y., Verspaget, H.W., Biemond, I., Sier, C.F., Lamers, C.B. and Veenendaal, R.A. (1996) Gastric mucosal plasminogen activators in Helicobacter pylori infection. Dig Dis Sci 41, 1577-82.
Hamlet, A.K., Erlandsson, K.I., Olbe, L., Svennerholm, A.M., Backman, V.E. and Pettersson, A.B. (1995) A simple, rapid, and highly reliable capsule-based 14C urea breath test for diagnosis of Helicobacter pylori infection. Scand J Gastroenterol 30, 1058-63.
Haruma, K., Okamoto, S., Sumii, K., Yoshihara, M., Tari, A., Teshima, H., Shimamoto, T., Takehara, Y., Tanaka, S., Yamamoto, G. and et al. (1992) Helicobacter pylori infection and gastroduodenal disease: a comparison of endoscopic findings, histology, and urease test data. Hiroshima J Med Sci 41, 65-70.
Hazell, S.L., Evans, D.J., Jr. and Graham, D.Y. (1991) Helicobacter pylori catalase. J Gen Microbiol 137 ( Pt 1), 57-61.
Huang, Q., Lan, X., Tong, T., Wu, X., Chen, M., Feng, X., Liu, R., Tang, Y. and Zhu, Z. (1996) Dot-immunogold filtration assay as a screening test for syphilis. J Clin Microbiol 34, 2011-3.
Kassa, E., Tsega, E. and Gebre, W. (1996) Comparison of diagnostic methods for detection of Helicobacter pylori. East Afr Med J 73, 239-41.
Katsuragi, K., Noda, A., Tachikawa, T., Azuma, A., Mukai, F., Murakami, K., Fujioka, T., Kato, M. and Asaka, M. (1998) Highly sensitive urine-based enzyme-linked immunosorbent assay for detection of antibody to Helicobacter pylori. Helicobacter 3, 289-95.
Kawanishi, M., Fukuda, S., Kawaguchi, H., Kohmoto, K., Haruma, K. and Kajiyama, G. (1995) Significance of rapid urease test for identification of Helicobacter pylori in comparison with histological and culture studies. J Gastroenterol 30, 16-20.
Kim, J.M., Kim, J.S., Jung, H.C., Song, I.S. and Kim, C.Y. (2000) Apoptosis of human gastric epithelial cells via caspase-3 activation in response to Helicobacter pylori infection: possible involvement of neutrophils through tumor necrosis factor alpha and soluble Fas ligands. Scand J Gastroenterol 35, 40-8.
Kohler, G. and Milstein, C. (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256, 495-7.
Lai, C.H., Kuo, C.H., Chen, Y.C., Chao, F.Y., Poon, S.K., Chang, C.S. and Wang, W.C. (2002) High prevalence of cagA- and babA2-positive Helicobacter pylori clinical isolates in Taiwan. J Clin Microbiol 40, 3860-2.
Laine, L., Estrada, R., Trujillo, M., Knigge, K. and Fennerty, M.B. (1998) Effect of proton-pump inhibitor therapy on diagnostic testing for Helicobacter pylori. Ann Intern Med 129, 547-50.
Leung, W., Chan, P., Bosgoed, F., Lehmann, K., Renneberg, I., Lehmann, M. and Renneberg, R. (2003) One-step quantitative cortisol dipstick with proportional reading. J Immunol Methods 281, 109-18.
Lim, J.W., Kim, H. and Kim, K.H. (2003) Cell adhesion-related gene expression by Helicobacter pylori in gastric epithelial AGS cells. Int J Biochem Cell Biol 35, 1284-96.
Luzza, F., Imeneo, M., Maletta, M., Paluccio, G., Nistico, S., Perticone, F., Foca, A. and Pallone, F. (1998) Suggestion against an oral-oral route of transmission for Helicobacter pylori infection: a seroepidemiological study in a rural area. Dig Dis Sci 43, 1488-92.
Malfertheiner, P., Enrique Dominguez-Munoz, J., Heckenmuller, H., Neubrand, M., Fischer, H.P. and Sauerbruch, T. (1996) Modified rapid urease test for detection of Helicobacter pylori infection. Eur J Gastroenterol Hepatol 8, 53-6.
Mao, S.a.D.F. (1984) Ehancement of limiting dilution in cloning mouse myeloma and spleen hybridomas by human low density lipoprotein. J. Immunol. Meth. 75, 309-316.
Marshall, B.J. and Warren, J.R. (1984) Unidentified curved bacilli in the stomach of patients with gastritis and peptic ulceration. Lancet 1, 1311-5.
Mendoza, L.G., McQuary, P., Mongan, A., Gangadharan, R., Brignac, S. and Eggers, M. (1999) High-throughput microarray-based enzyme-linked immunosorbent assay (ELISA). Biotechniques 27, 778-80, 782-6, 788.
Miki, H., Kobayashi, S., Harada, H., Yamanoi, Y., Uraoka, T., Sotozono, M. and Ohmori, M. (2001) Early stage gastric MALT lymphoma with high-grade component cured by Helicobacter pylori eradication. J Gastroenterol 36, 121-4.
Murakami, K., Sato, R., Okimoto, T., Nasu, M., Fujioka, T., Kodama, M. and Kagawa, J. (2003) Efficacy of triple therapy comprising rabeprazole, amoxicillin and metronidazole for second-line Helicobacter pylori eradication in Japan, and the influence of metronidazole resistance. Aliment Pharmacol Ther 17, 119-23.
Neoh, S.H., Gordon, C., Potter, A. and Zola, H. (1986) The purification of mouse monoclonal antibodies from ascitic fluid. J Immunol Methods 91, 231-5.
Ohkuma, K., Okada, M., Murayama, H., Seo, M., Maeda, K., Kanda, M. and Okabe, N. (2000) Association of Helicobacter pylori infection with atrophic gastritis and intestinal metaplasia. J Gastroenterol Hepatol 15, 1105-12.
Parsonnet, J., Friedman, G.D., Vandersteen, D.P., Chang, Y., Vogelman, J.H., Orentreich, N. and Sibley, R.K. (1991) Helicobacter pylori infection and the risk of gastric carcinoma. N Engl J Med 325, 1127-31.
Parsonnet, J., Hansen, S., Rodriguez, L., Gelb, A.B., Warnke, R.A., Jellum, E., Orentreich, N., Vogelman, J.H. and Friedman, G.D. (1994) Helicobacter pylori infection and gastric lymphoma. N Engl J Med 330, 1267-71.
Perri, F., Ghoos, Y., Hiele, M., Andriulli, A. and Rutgeerts, P. (1995) The urea breath test: a non-invasive clinical tool for detecting Helicobacter pylori infection. Ital J Gastroenterol 27, 55-63.
Quiros, A., Quiros, E., Gonzalez, I., Bernal, M.C., Piedrola, G. and Maroto, M.C. (1994) Helicobacter pylori seroepidemiology in risk groups. Eur J Epidemiol 10, 299-301.
Sathar, M.A., Gouws, E., Simjee, A.E. and Mayat, A.M. (1997) Seroepidemiological study of Helicobacter pylori infection in South African children. Trans R Soc Trop Med Hyg 91, 393-5.
Sebastian, M., Chandran, V.P., Elashaal, Y.I. and Sim, A.J. (1995) Helicobacter pylori infection in perforated peptic ulcer disease. Br J Surg 82, 360-2.
Stadtlander, C.T. and Stutzenberger, F.J. (1995) Adaptation of the [13C]urea breath test as a noninvasive method for detection of Helicobacter pylori infection in squirrel monkeys (Saimiri spp.). Lab Anim Sci 45, 239-43.
Sullivan, T., Ashbury, F.D., Fallone, C.A., Naja, F., Schabas, R., Hebert, P.C., Hunt, R. and Jones, N. (2004) Helicobacter pylori and the prevention of gastric cancer. Can J Gastroenterol 18, 295-302.
Suzuki, N., Wakasugi, M., Nakaya, S., Kokubo, N., Sato, M., Kajiyama, H., Takahashi, R., Hirata, H., Ezure, Y., Fukuda, Y. and Shimoyama, T. (2002a) Catalase, a specific antigen in the feces of human subjects infected with Helicobacter pylori. Clin Diagn Lab Immunol 9, 784-8.
Suzuki, N., Wakasugi, M., Nakaya, S., Okada, K., Mochida, R., Sato, M., Kajiyama, H., Takahashi, R., Hirata, H., Ezure, Y., Koga, Y., Fukuda, Y. and Shimoyama, T. (2002b) Production and application of new monoclonal antibodies specific for a fecal Helicobacter pylori antigen. Clin Diagn Lab Immunol 9, 75-8.
Urakami, Y. and Sano, T. (2001) Endoscopic duodenitis, gastric metaplasia and Helicobacter pylori. J Gastroenterol Hepatol 16, 513-8.
Valkonen, K.H., Ringner, M., Ljungh, A. and Wadstrom, T. (1993) High-affinity binding of laminin by Helicobacter pylori: evidence for a lectin-like interaction. FEMS Immunol Med Microbiol 7, 29-37.
van de Wouw, B.A., de Boer, W.A., Jansz, A.R., Staals, A.P. and Roymans, R.T. (1995) Serodiagnosis of Helicobacter pylori infection: an evaluation of a commercially available ELISA-IgG. Neth J Med 47, 272-7.
Voigt, A. (1989) [Monoclonal antibodies in research and clinical medicine]. Kinderarztl Prax 57, 479-87.
Walsh, J.H. and Peterson, W.L. (1995) The treatment of Helicobacter pylori infection in the management of peptic ulcer disease. N Engl J Med 333, 984-91.
Warchol, J.B. and Brelinska, R. (1980) Some aspects of labelling of proteins with colloidal gold. Acta Med Pol 21, 413-4.
Warren, J.R.a.M., B.J. (1983) Unidentified curved bacilli on gastric epithelium in active chronic gastritis. Lancet 1, 1273-5.
Yahiro, K., Niidome, T., Hatakeyama, T., Aoyagi, H., Kurazono, H., Padilla, P.I., Wada, A. and Hirayama, T. (1997) Helicobacter pylori vacuolating cytotoxin binds to the 140-kDa protein in human gastric cancer cell lines, AZ-521 and AGS. Biochem Biophys Res Commun 238, 629-32.
Yang, S.J.a.S., J. T. Mao. (1999) A simple HPLC Purification Procedure for Porcine Plasma Haptoglobin. J. Chromatagr. B. 731, 395-402.