[논문]Antigenic Proteins of Helicobacter pylori of Potential Diagnostic Value

Khalilpour, Akbar; Santhanam, Amutha; Lee, Chun Wei; Saadatnia, Geita; Velusamy, Nagarajan; Osman, Sabariah; Mohamad, Ahmad Munir; Noordin, Rahmah

doi:10.7314/apjcp.2013.14.3.1635

Antigenic Proteins of Helicobacter pylori of Potential Diagnostic Value 원문보기

Asian Pacific journal of cancer prevention : APJCP, v.14 no.3, 2013년, pp.1635 - 1642

Khalilpour, Akbar (Institute for Research in Molecular Medicine, Universiti Sains Malaysia) , Santhanam, Amutha (Institute for Research in Molecular Medicine, Universiti Sains Malaysia) , Lee, Chun Wei (Institute for Research in Molecular Medicine, Universiti Sains Malaysia) , Saadatnia, Geita (Institute for Research in Molecular Medicine, Universiti Sains Malaysia) , Velusamy, Nagarajan (Hospital Seberang Jaya) , Osman, Sabariah (Institute for Research in Molecular Medicine, Universiti Sains Malaysia) , Mohamad, Ahmad Munir (Institute for Research in Molecular Medicine, Universiti Sains Malaysia) , Noordin, Rahmah (Institute for Research in Molecular Medicine, Universiti Sains Malaysia)

Abstract ▼ AI-Helper

Helicobacter pylori antigen was prepared from an isolate from a patient with a duodenal ulcer. Serum samples were obtained from culture-positive H. pylori infected patients with duodenal ulcers, gastric ulcers and gastritis (n=30). As controls, three kinds of sera without detectable H. pylori IgG antibodies were used: 30 from healthy individuals without history of gastric disorders, 30 from patients who were seen in the endoscopy clinic but were H. pylori culture negative and 30 from people with other diseases. OFF-GEL electrophoresis, SDS-PAGE and Western blots of individual serum samples were used to identify protein bands with good sensitivity and specificity when probed with the above sera and HRP-conjugated anti-human IgG. Four H. pylori protein bands showed good (${\geq}$ 70%) sensitivity and high specificity (98-100%) towards anti-Helicobacter IgG antibody in culture-positive patients sera and control sera, respectively. The identities of the antigenic proteins were elucidated by mass spectrometry. The relative molecular weights and the identities of the proteins, based on MALDI TOF/TOF, were as follows: CagI (25 kDa), urease G accessory protein (25 kDa), UreB (63 kDa) and proline/pyrroline-5-carboxylate dehydrogenase (118 KDa). These identified proteins, singly and/or in combinations, may be useful for diagnosis of H. pylori infection in patients.

주제어

AI 본문요약
AI-Helper

* AI 자동 식별 결과로 적합하지 않은 문장이 있을 수 있으니, 이용에 유의하시기 바랍니다.

문제 정의

To the best of our knowledge, this is the first report to identify CagI, UreG, and proline/pyrroline-5-carboxylate dehydrogenase as H. pylori antigenic proteins. CagI has been described as a putative surface protein based on its tendency to interact with integrin (Jiménez-Soto LF, 2009).

제안 방법

The isolates were identified by colony morphology, negative Gram staining and positive reactions in biochemical tests for urease, catalase and oxidase. In addition, PCR using published primers was performed to confirm the presence of the CagA and VacA genes.
pylori antibodies when tested with individual patients’ serum samples, and the specificity of each band was determined based on the reactivity with the individual control serum samples from Groups II, III and IV (Table 2).

대상 데이터

pylori IgG antibodies. The group comprised two patients with duodenal ulcer, five patients with gastric ulcer, 20 patients with gastritis and three patients with normal scope findings (n=30). Group II patients were healthy individuals without any history of gastric disorders (n=30).
The samples were analyzed using an ABI 4800 MALDI TOF-TOF mass spectrometer at the Protein and Proteomics Centre, Faculty of Biological Sciences, National University of Singapore. GPS Explorer^TM software Version 3.

이론/모형

An in-gel digestion of the gel piece containing the protein band was performed using the method of O’Connell and Stalts (1997).
Finally, the sample was vortexed well and centrifuged for 15 min at 10,000 X g. The protein content of the supernatant was determined using the RCDC method (BioRad, USA).

성능/효과

pylori has been reported in different geographic regions and within a population. A study of 78 H. pylori isolates from three major ethnic groups in Malaysia using multilocus sequence typing (MLST) of seven housekeeping genes and comparison with global H. pylori data showed that the isolates can be differentiated into three populations i.e. hpEastAsia, hpAsia2 and hpEurope. Interestingly, the Malaysian Indian and Malay isolates formed a new subpopulation within hpAsia2 i.
In conclusion, four H. pylori infection markers identified in this study showed good sensitivity and high specificity, and three of them have not previously been reported to be antigenic. Recombinant forms of these proteins, either singly or in combinations, are potentially useful for development of a diagnostic test for H.

후속연구

However, the identities of the antigens used in commercial kits are usually not disclosed. Thus, mixtures of various combinations of antigens identified in this study may be tested for their usefulness in detection of H. pylori infection.

참고문헌 (73)

Andersen LP, Espersen F (1992). Immunoglobulin G antibodies to Helicobacter pylori in patients with dyspeptic symptoms investigated by the western immunoblot technique. J Clin Microbiol, 30, 1743-51.
Alam J, Maiti S, Ghosh P, et al (2012). Significant association of the dupA gene of Helicobacter pylori with duodenal ulcer development in a South-east Indian population. J Med Microbiol, 61, 1295-302.

상세보기
Baik SC, Kim KM, Song SM, et al (2004). Proteomic analysis of the sarcosine-insoluble outer membrane fraction of Helicobacter pylori strain 26695. J Bacteriol, 186, 949-55.

상세보기
Benoit S, Mehta N, Wang G, Gatlin M, Maier RJ (2004). Requirement of hydD, hydE, hypC and hypE genes for hydrogenase activity in Helicobacter pylori. Microb Pathog, 36, 153-7.

상세보기
Bergey B MP, Peacock J, Megraud F (2003). What is the role of serology in assessing Helicobacter pylori eradication. Alimentary pharmacology & therapeutics, 18, 635-9.

상세보기
Besra GS, Kremer L, Nampoothiri KM, et al (2001). Biochemical characterization of acyl carrier protein (AcpM) and malonyl-CoA: AcpM transacylase (mtFabD), two major components of Mycobacterium tuberculosis fatty acid synthase II. J Biological Chemistry, 76, 27967-74.
Chan CH, Ko CC, Chang JG, et al (2006). Subcellular and functional proteomic analysis of the cellular responses induced by Helicobacter pylori. Mol Cell Proteomics, 4, 702-13.
Chambers MG, Pyburn TM, et al (2013). Structural analysis of the oligomeric states of Helicobacter pylori VacA toxin. J Mol Biol, 425, 524-35.

상세보기
Chisholm SA, Owen RJ (2008). Application of polymerase chain reaction-based assays for rapid identification and antibiotic resistance screening of Helicobacter pylori in gastric biopsies. Diagn Microbiol Infect Dis, 61, 67-71.

상세보기
Chomvarin C, Ottiwet O, Hahnvajanawong C, Intapan PM, Wongwajana S (2009). Seroreactivity to specific antigens of Helicobacter pylori infection is associated with an increased risk of the dyspeptic gastrointestinal diseases. Int J Infect Dis, 13, 647-54.

상세보기
Crabtree JE, Shallcross TM, Heatley RV, Wyatt JI (1991). Evaluation of a commercial ELISA for serodiagnosis of Helicobacter pylori infection. J Clin Pathology, 44, 326-8.

상세보기
Cremonini F, Gabrielli M, Gasbarrini G, Pola P, Gasbarrini A (2004). The relationship between chronic H. pylori infection, CagA seropositivity and stroke: meta-analysis. Atherosclerosis, 173, 253-9.

상세보기
Evans DJJ, Evans DG, Graham DY, Klein PD (1989). A sensitive and specific serologic test for detection of Campylobacter pylori infection. Gastroenterology, 96, 1004-8.
Flores-luna L, Camorlinga-Ponce M, Hernendez-Suarez G, et al (2013). The utility of serologic tests as biomarkers for Helicobacter pylori-associated precancerous lesions and gastric cancer varies between Latin American countries. Cancer Causes Control, 24, 241-8.

상세보기
Faulde M, Putzker M, Mertes T, Sobe D (1991). Evaluation of an immunofluorescence assay for specific detection of immunoglobulin G antibodies directed against Helicobacter pylori, and antigenic cross-reactivity between H. pylori and Campylobacter jejuni. J Clin Microbiol, 29, 323-7.
Franceschi F, Gasbarrini A (2007). Helicobacter pylori and extragastric diseases. Best Pract Res Clin Gastroenterol, 21, 325-34.

상세보기
Galmiche A, Rassow J, Doye A, et al (2000). The N-terminal 34 kDa fragment of Helicobacter pylori vacuolating cytotoxin targets mitochondria and induces cytochrome c release. EMBO J, 19, 6361-70.

상세보기
Ge R, Sun X, Gu Q, et al (2007). A proteomic approach for the identification of bismuth-binding proteins in Helicobacter pylori. J Biol Inorg Chem, 12, 831-42.

상세보기
Giannakis M, Chen SL, Karam SM, Engstrand L, Gordon JI (2008). Helicobacter pylori evolution during progression from chronic atrophic gastritis to gastric cancer and its impact on gastric stem cells. Proc Natl Acad Sci USA, 105, 4358-63.

상세보기
Glassman MS, Dallal S, Berezin SH, et al (1990). Helicobacter pylori-related gastroduodenal disease in children. Diagnostic utility of enzyme-linked immunosorbent assay. Dig Dis Sci, 35, 993-7.

상세보기
Graham DY, Evans DJ, Jr, Peacock J, Baker JT, Schrier WH (1996). Comparison of rapid serological tests (FlexSure HP and QuickVue) with conventional ELISA for detection of Helicobacter pylori infection. Am J Gastroenterology, 91, 942-8.
Haas G, Karaali G, Ebermayer K, et al (2002). Immunoproteomics of Helicobacter pylori infection and relation to gastric disease. Proteomics, 2, 313-24.

상세보기
Hambers MG, Pyburn TM, Gonzalez-rivera C, et al (2013). Structural analysis of the oligomeric states of Helicobacter pylori VacA toxin. J Mol Biol, 425, 524-35.

상세보기
Inagaki E, Ohshima N, Takahashi H, et al (2006). Crystal structure of Thermus thermophilus Delta1-pyrroline-5-carboxylate dehydrogenase. J Mol Biol, 362, 490-501.

상세보기
Jensen AKV, Andersen LP, Gaarslev K, Wachmann CH (1993). Comparison of four second generation kits for detection of IgG antibodies against Helicobacter pylori in adults. Zentralblatt fur Bakteriologie, 280, 221-6.

상세보기
Jimenez-Soto LF KS, Sewald X, Ertl C, et al (2009). Helicobacter pylori type IVsecretion apparatus exploits b1 integrin in anovel RGD-independent manner. PLoS Pathog, 5,1000684.

상세보기
Jungblut PR, Bumann D, Haas G, et al (2000). Comparative proteome analysis of Helicobacter pylori. Mol Microbiol, 36,710-25.

상세보기
Keenan J OJ, Domigan N, Potter H, et al (2000). Immune response to an 18-kilodalton outer membrane antigen identifies lipoprotein 20 as a Helicobacter pylori vaccine candidate. Infect Immun, 68, 3337-43.

상세보기
Keenan J, Oliaro J, Domigan N, et al (2000). Immune response to an 18-kilodalton outer membrane antigen identifies lipoprotein 20 as a Helicobacter pylori vaccine candidate. Infect Immun, 68, 3337-43.

상세보기
Khanna B, Cutler A, Israel NR, et al (1998). Use caution with serologic testing for Helicobacter pylori infection in children. J Infectious Diseases, 178, 460-5.

상세보기
Krah A, Miehlke S, Pleissner KP, et al (2004). Identification of candidate antigens for serologic detection of Helicobacter pylori-infected patients with gastric carcinoma. Int J Cancer, 108, 456-63.

상세보기
Krah A MS, Pleissner KP, Zimny-Arndt U, et al (2004). Identification of candidate antigens for serologic detection of Helicobacter pylori-infected patients with gastric carcinoma. Int J Cancer, 108, 456-63.

상세보기
Kwok T, Zabler D, Urman S, et al (2007). Helicobacter exploits integrin for type IV secretion and kinase activation. Nature, 449, 862-6.

상세보기
Leung WK, Ng EKW, Chan FKL, Chung SCS, Sung JJY (1999). Evaluation of three commercial enzyme-linked immunosorbent assay kits for diagnosis of Helicobacter pylori in Chinese patients. Diagnostic Microbiology and Infectious Disease, 34, 13-17.

상세보기
Lin Y-F, Chen C-Y, Tsai M-H, et al (2007). Duodenal ulcerrelated antigens from Helicobacter pylori. Immunoproteome and protein microarray approaches. Molecular and Cellular Proteomics, 6, 1018-26.

상세보기
Lochhead P, El-Omar EM (2007). Helicobacter pylori infection and gastric cancer. Best Pract Res Clin Gastroenterol, 21, 281-97.

상세보기
Malaty HM (2007). Epidemiology of Helicobacter pylori infection. Best Pract Res Clin Gastroenterol, 21, 205-14.

상세보기
Manes G BA, Iaquinto G, Ricci C, et al (2001). Accuracy of the stool antigen test in the diagnosis of Helicobacter pylori infection before treatment and in patients on omeprazole therapy. Aliment Pharmacol Ther, 15, 73-9.

상세보기
Manes G, Balzano A, Iaquinto G, et al (2001). Accuracy of stool antigen test in posteradication assessment of Helicobacter pylori infection. Dig Dis Sci, 46, 2440-4.

상세보기
Manes G, Zanetti MV, Piccirillo MM, et al (2005). Accuracy of a new monoclonal stool antigen test in post-eradication assessment of Helicobacter pylori infection: comparison with the polyclonal stool antigen test and urea breath test. Dig Liver Dis, 37, 751-5.

상세보기
Mao YF, Yan J (2004). Construction of prokaryotic expression system of ureB gene from a clinical Helicobacter pylori strain and identification of the recombinant protein immunity. World J Gastroenterology, 10, 977-84.
Marchildon PA, Ciota LM, Zamaniyan FZ, Peacock JS, Graham DY (1996). Evaluation of three commercial enzyme immunoassays compared with the 13C urea breath test for detection of Helicobacter pylori infection. J Clin Microbiology, 34, 1147-52.
Meijer BC, Thijs JC, Kleibeuker JH, van Zwet AA, Berrelkamp RJP (1997). Evaluation of eight enzyme immunoassays for detection of immunogloblin G against Helicobacter pylori. J Clinical Microbiology, 35, 292-4.
Michetti P KC, Kotoff K, Porta N, et al (1999). Oral immunization with urease and Escherichia coli heat-labile enterotoxin is safe and immunogenic in Helicobacter pyloriinfected adults. Gastroenterology, 116, 804-12.

상세보기
Mini R, Bernardini G, Salzano AM, et al (2006). Comparative proteomics and immunoproteomics of Helicobacter pylori related to different gastric pathologies. J Chromatography B, Analytical Technologies in the Biomedical and Life Sciences, 833, 63-79.

상세보기
Miwa H, Go MF, Sato N (2002). H. pylori and gastric cancer: the Asian enigma. Am J Gastroenterol, 97, 1106-12.

상세보기
Miwa H, Akamatsu S, Tachikawa T, et al (2001). On-site diagnosis of H. pylori infection by urine. Diagn Microbiol Infect Dis, 39, 95-7.

상세보기
Miwa H, Kikuchi S, Ohtaka K, et al (2000). Insufficient diagnostic accuracy of imported serological kits for Helicobacter pylori infection in Japanese population. Diagnostic Microbiology Infectious Disease, 36, 95-9.

상세보기
Mobley HL (1996). The role of Helicobacter pylori urease in the pathogenesis of gastritis and peptic ulceration. Aliment Pharmacol Ther, 10, 157-64.

상세보기
Noto JM, Gaddy JA, Lee JY, et al (2013). Iron deficiency accelerates Helicobacter pylori-induced carcinogenesis in rodents and humans. J Clin Invest, 123, 479-92.

상세보기
O'Connell KL, Stalts JT (1997). Identification of mouse liver proteins on two-dimensional electrophoresis gel by matrixassociated larser desorption/ ionization mass spectrometry of in situ enzymatic digests. Electrophoresis, 18, 349-59.

상세보기
Obata Y, Kikuchi S, Miwa H, et al (2003). Diagnostic accuracy of serological kits for Helicobacter pylori infection with the same assay system but different antigens in a Japanese patient population. J Medical Microbiology, 52, 889-92.

상세보기
Ohtsuka Y, Shimada T, Watanabe N, et al (1997). Analysis of the vacA gene sequences among Helicobacter pylori strains isolated from Japanese patients. Gastroenterology, 112, 243.
Park J-W, Song J-Y, Lee S-G, et al (2006). Quantitative analysis of representative proteome components and clustering of Helicobacter pylori clinical strains. Helicobacter, 11, 533-43.

상세보기
Parsons TB, Ghellamallah C, Male L, Spencer N, Grainger RS (2011). Regioselective dibromination of methyl indole-3-carboxylate and application in the synthesis of 5,6-dibromoindoles. Org Biomol Chem, 9, 5021-3.

상세보기
Pelerito A, Oleastro M, Lopes AI, et al (2006). Evaluation of rapid test Assure Helicobacter pylori for diagnosis of H. pylori in pediatric population. J Microbiol Methods, 66, 331-5.

상세보기
Rieder G, Fischer W, Haas R (2005). Interaction of Helicobacter pylori with host cells: function of secreted and translocated molecules. Curr Opin Microbiol, 8, 67-73.

상세보기
Robinson K, Argent RH, Atherton JC (2007). The inflammatory and immune response to Helicobacter pylori infection. Best Pract Res Clin Gastroenterol, 21, 237-59.

상세보기
Sasidharan S, Uyub AM (2009). Antibody response to Helicobacter pylori excretory antigen and the cross reaction study. J Immunoassay Immunochem, 30, 70-81.
Schumann C, Triantafilou K, Rasche FM, et al (2006). Serum antibody positivity for distinct Helicobacter pylori antigens in benign and malignant gastroduodenal disease. IJMM, 296, 223-8.
Shevchenko A, Wilm M, Vorm O, Mann M (1996). Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal Chem, 68, 850-8.

상세보기
Simor AE, Lin E, Saibil F, et al (1996). Evaluation of enzyme immunoassay for detection of salivary antibody to Helicobacter pylori. J Clin Microbiol, 34, 550-3

상세보기
Tay Y, Mitchell H, Dong Q, et al (2009). Population structure of Helicobacter pylori among ethnic groups in Malaysia: recent acquisition of the bacterium by the Malay population. BMC Microbiology, 9, 1-13.

상세보기
Torres J, Camorlinga-Ponce M, Perez-Perez G, Munoz L, Munoz O (2002). Specific serum immunoglobulin G response to urease and CagA antigens of Helicobacter pylori in infected children and adults in a country with high prevalence of infection. Clin Diagn Lab Immunol, 9, 97-100.

상세보기
Utt M, Nilsson I, Ljungh A, Wadstrom T (2002). Identification of novel immunogenic proteins of Helicobacter pylori by proteome technology. J Immunological Methods, 259, 1-10.

상세보기
Vaira D, Vakil N (2001). Blood, urine, stool, breath, money, and Helicobacter pylori. Gut, 48, 287-9.

상세보기
Voland P, Weeks DL, Vaira D, Prinz C, Sachs G (2002). Specific identification of three low molecular weight membraneassociated antigens of Helicobacter pylori. Alimentary Pharmacology & Therapeutics, 16, 533-44.

상세보기
Widmer M, de Korwin JD, Aucher P, et al (1999). Performance of native and recombinant antigens for diagnosis of Helicobacter pylori infection. Eur J Clin Microbiol Infect Dis, 18, 823-6.

상세보기
Wu M-S, Chow L-P, Lin J-T, Chiou S-H (2008). Proteomic identification of biomarkers related to Helicobacter pylori-associated gastroduodenal disease: challenges and opportunities. J Gastroenterol Hepatol, 23, 1657-61.

상세보기
Zhang C, Xu S, Xu D (2012). Risk assessment of gastric cancer caused by Helicobacter pylori using CagA sequence markers. PLoS One, 7, 36844.

상세보기
Zhang X, Shen W, Lu Y, et al (2010). Expression of UreB and HspA of Helicobacter pylori in silkworm pupae and identification of its immunogenicity. Mol Biol Rep, 38, 3173-80.
Zambelli B, Turano P, Musiani F, Neyroz P, Ciurli S (2009). Zn2+-linked dimerization of UreG from Helicobacter pylori, a chaperone involved in nickel trafficking and urease activation. Proteins: Structure, Function, and Bioinformatics, 74, 222-39.
Zheng J, Xiao- Hong T, Huang A-L, Wang P-L (2002). A study of recombinant Protective H. pylori antigens. World J Gastroenterology, 8, 308-11.

표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관 관리번호, 논문명, 대등논문명, 저자 , 저널/프로시딩명, 발행기관, 발행년, 발행언어, 권, 호, 시작페이지, 끝페이지, ISBN, ISSN, 주제분야, 키워드, 초록(한글), 초록(영문), 저자(소속기관)
저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증