BAB VI SIMPULAN DAN SARAN
6.2 Saran
1. Data penelitian ini dapat dimanfaatkan sebagai referensi pertama tentang kaitan polimorfisme -238G/A dan -308G/A dengan kejadian PPOK pada orang dengan riwayat merokok di Indonesia.
2. Penelitian tentang ekspresi alel -308A gen TNFα pada penderita
PPOK terhadap kadar TNFα di darah penderita PPOK perlu dilanjutkan. Analisis lebih lanjut pada sekuens disekitar posisi -
308 promotor gen TNFα dan identifikasi protein yang berkaitan
dengan sekuens DNA yang mencakup posisi -308 akan memberikan informasi yang bermanfaat dalam upaya karakterisasi promotor gen dan kaitannya terhadap faktor protektif kejadian PPOK.
3. Penelitian tentang polimorfisme gen TNFα -308G/A dan - 238G/A dan interaksi dengan kandidat gen lainnya kaitannya dengan kejadian PPOK perlu dilanjutkan.
DAFTAR KEPUSTAKAAN
Agusti, A., 2007. Systemic Effectof Chronic Obstructive Pulmonary Disease : What We Know and What We Don’t Know
(but should). AmThoracSoc 4: 522 – 525.
Andreassen, H. andVestbo, J., 2003. Chronic Obstructive Pulmonary Disease as a systemic disease : an epidemiological
perspective. EurRespir J 22 : 2 – 4.
Baratawidjaja, K.G.danRengganis, I., 2009. Sitokin.Dalam : Baratawidjaja, K.GdanRengganis, I.Imunologi
dasar. FakultasKedokteranUniversitas Indonesia, Jakarta.
Barnes, P.J., 2003. Inflammation.In :Caverley, P.M.A.eds., Chronic Obstructive Pulmonary Disease. Arnold: 139 – 138 .
Bayley, J.P., de Rooij, H., van den Elsen, P.J., Huizinga, T.W., Verweij, C.L.,2001. Functional analysis of linker scan mutants
spanning the -376, -308, -244 and -238 polymorphis sites of the TNF alpha promoter. Cytokine 14:316-323
Brantly, M., Nukiwa, T., and Crystal, R.G., 1988. Molecular basis of alpha antitrypsin deficiency. Am J Med 84 : 13 – 31.
Burhan, E., dan Yunus, F., 2001.Perubahan Faal paru pada orang tua. J.Respir Indonesia 21:160-165
Burrows, B., Boom, J., Traver, G.,1987.The Course and prognosis of different forms of Chronic airways obstruction in a sample
from the general population. N Engl J Med 317:1209 – 1214.
Burmeister, M., 1999. Basic Concepts in the Study of Diseases with ComplexGenetics.Biol Psychiatry 45:522-532
Candly, dan Tarigan, A.P., 2010. ProfilpenderitaPenyakitParu ObstruksiKronikEksaserbasi di RSUP.H.Adammalikpada tahun 2009. FakultasKedokteran, Universitas Sumatera Utara, Medan.
Castaldi, P. J.,Cho,M.H, Chon, M., Langerman,F., Moran,S., Tarragonna, N., 2010.The COPD genetic association
compendium :a comprehensive online database of COPD geneticassociations.HumanMolecular Genetics 19 (3 ) : 526-534.
Chierakul, N., Wongwisutikul, P., Vejbyasea, P., Chotvilaiwan, K., 2005. Tumor necrosis factor-α gene promoter polymorphism is not associated with smoking related COPD in Thailand.
Respirol
Cho M.H., and Silverman E.K., 2009. Genetic And Racial, Ethnic, And Gender Characteristics Of COPD. In: Atlas of Chronic
Obstructive Pulmonary Disease.Springer :41 – 63.
Chung, K.F., 2002. Cytokines.In : Barnes, P.J., eds.,Asthma and
COPD.Basic mechanisms and clinical management.
AcademicPress : 261 – 271.
Dawud, Y., 2004. Occupational & environmental lung disorder.J RespirIndonesia 24 : 126 – 33.
DeMeo, D.L., Campbell, E.J., Barker, A.F., Brantly, M.L., Eden, E., Mac Elvaney,M.G.,2008. IL10 polymorphisms are
associated with airflow obstruction in severe Alpha1- antitrypsindeficiency.Am JRespir Cell MolBiol38 : 114 – 120.
DepartemenKesehatanRepublik Indonesia., 1999. Metodologi PenelitianKesehatan. PenuntunLatihanMetodaPenelitian. BadanPenelitiandanPengembanganKesehatan,Pusat PenelitianPenyakitTidakMenular. Jakarta.
De Vries,R.R., and Van Rood,J.J.,1992.Immunogenetic and Diseases in The Genetic Basis of Common Diseases.University Press
Inc, New York:92-144
Fatchiyah, Arumingtyas, E.L., Widyarti, S., Rahayu, S.,2011. Biologi Molekular.Prinsip Dasar Analisis.Penerbit Erlangga.Jakarta
Fujita, M., Shannon,J.M., Irvin, C.G., Fagan,K.A., Cool,C., Augustin, A., 2001. Overexpression of tumor necrosis factor α
produces an increase in lung volumes and pulmonary hypertension. Am J Physiol LungCell MolPhysiol280 : L39 – L49.
Forum of International Respiratory Societies (FIRS)., 2010. The year of the lung. Available from: http://www.2010yearofthe
lung.org. [Accessed on 1 Mei 2010].
Gadgil, A., and Duncan, S.R., 2008. Role of T lymphocytes and pro inflammatory mediators in the pathogenesis of chronic
obstructive pulmonary disease. Int JChron Obstruct Pulmon Dis 3 (4) : 531 – 541 .
Gauderman, W.J.,McConnel, R., Gilliland,F., London, S., Thomas, D., Avol,E., 2000. Association between air pollution and
lung function growth in Southern California children. Am J RespirCrit Care Med 162 : 1383 – 90.
Gingo, M.R., Silviera, F.J., Miller, Y.E., Friedlander, A.L., Cosgrove, G.P., Chan, E.D., 2008. Tumour necrosis factor gene polymorphisms are associated with COPD. EurRespirJ 31 : 1005 – 1012
Girodet P.O., and Raherison C., 2009.Epidemiology of COPD. Eur Respir Rev 18,114:213-221.
Global Initiative for Chronic Obstructive Lung Disease (GOLD)., 2011.
Pocket guide to COPD diagnosis, management, and
prevention. GOLD, Medical Communication Resources, Inc.
Gold, D.R.,Wang, X., Wipij,D., Speizer,F.E., Ware, J.H., Dockery,D.W., 1996. Effect of cigarette smoking on the pulmonary function
in adolescent boys and girls. N Engl J Med 335 : 931 – 937
Grana,L.M., Carnota,J.G.R., 2001.Tumor Necrosis Factor:genetic, cell action mechanism and involvement in Inflammation. Allergy Immunol.Clin16:140-149
Gruen J.R., Weissman,S.M.,1997. Evolving views of the Major histocompatibility complex. Blood 90:4252-4265
Hajeer, A.H., and Hutchinson, I.V., 2000. TNF-a gene polymorphism: clinical and biological implications. MicroscRes Tech 50 :
216 – 228.
Halbert R.J., Natoli,J.L., Gano,A., Badamqarav, E., Buist, A.S., Mannino, D.M.,2006.Global burden of COPD:Systemic review and Meta-analysis.Eur Respir J 28 :523 – 532.
Han, M.K., Postma, D., Mannino,D.M., Giardino,N.D., Buist,S., Curtis,J.L.,2007. Gender and Chronic Obstructive Pulmonary Disease: Why it matters. Am J Respir Crit Care Med 176:1179 – 1184.
Haq, G., Han, W., and Kim, C., 2002. Urban air pollution.Management andpracticein Major and Megacities of Asia. UNEP / WHO.
Hegab, A.E., Sakamoto, T., Saitoh,W., Nomura, A., Ishii, Y., Morishima, Y., 2005. Polymorphisms ofTNFα, IL1β, andIL1RN genes in chronic obstructive pulmonary disease.
Biochemical and Biophysical Research Communications
329 (4) : 1246 – 1252 .
Hersh, C.P., Demeo, D.L., Lange,C., Litonjua,A.A., Reilly, J.J., Kwiatkowski,D., 2005. Attempted replication of reported chronic obstructive pulmonary disease candidate
gene associations. American Journal of Respiratory Cell and Molecular Biology 3 : 371-78
Higham,M.A. Pride,N.B., Alikhan, A., Morel, N.W., 2000. Tumor
Necrosis factorα gene promoter polymorphism in chronic
obstructive pulmonary disease. EurRespir J 15:282–284 . Higuchi,T.,Seki, N., Kamizono, S., Yamada, A., Kimura,A., Kato,H.,
1998. Polymorphism of the 5’-flanking region of the human tumor necrosis factor (TNF)-a gene in Japanese. Tissue Antigens 51: 605 – 612.
Hogg, J.C., 2002. Airway Pathology.In : Barnes, P.J. Eds. Asthma and
COPD.Basic mechanisms andclinicalmanagement.
Academic Press :57 – 66 .
Huang, S.L., Su C.H., and Chang,S.C.,1997. Tumor Necrosis Factor
aGenePolymorphism in Chronic Bronchitis. Am J RespirCrit
Care Med156 : 1436 –1439.
Huizinga, T.W., Westendorp, R.G., Bollen,E.L.,Keijsers,V.,Brinkman, B.M.,Langermans,J.A.,1997. TNF-a promoter polymorphisms, production and susceptibility to multiple sclerosis in different
groups of patients. J Neuroimmunol72 : 149 – 153.
Hu, G.P., Peng, G.Y., and Ran, P.X., 2007. Association of tumor necrosis factor alpha308 G/A gene promoter
polymorphism with the presence of chronicobstructive pulmonary disease: a metaanalysis. Chineese Journal of Tuberculosis and Respiratory Disease 30 (8) : 594 – 599. Ishii,T., 2000.Neither IL-1 β, IL-1 receptor antagonist, nor TNF- α polymorphisms are associated with susceptibility to COPD.
Association of gene polymorphisms of tumour necrosis factor-
α and interleukin-13 withchronic obstructive pulmonary disease in Hannationality in Beijing. Chinese medical journal 118 (7) : 541 – 547 .
Kroeger, K.M., Carville, K.S., Abraham, L.J., 1997. The -308 tumor necrosis factor-a promoter polymorphism effects transcription.
MolImmunol34 : 391 – 399.
Kurniawidjaja L.M., 2004. Peranan Variasi Genetik Pada gen TNFα
Posisi -308, SitokinTNFα, dan Sitokin IL10 terhadap Silikosis Pekerja Pabrik Semen Di Indonesia. Disertasi,Program
Doktor Ilmu Epidemiologi,Program Pasca Sarjana Fakultas Kesehatan Masyarakat,Universitas Indonesia.
Lange, P., 1992. Development and Prognosis of chronic obstructive pulmonary disease with special reference to the role of
tobacco smoking. Dan MedBull 39 : 30 – 48.
Louis, E., Franchimont, D., Piron,A., Gevaert,Y.,Schaaf,L.N.,Roland,S., 1998. Tumour necrosis factor (TNF) gene
polymorphism influences TNF-a production in
lipopolysaccharide (LPS)stimulated whole blood cell culture in healthy humans. ClinExpImmunol113 : 401 – 406.
Lokke, A., Lange, P., Scharling, H., Fabricius,P., Vestbo,J.,2006. Developing COPD : a 25 year follow up study of the general population.Thorax 61(11): 935-939
Mac Guire, W., Hill, A.V., Allsopp,C.E., Greenwood,B.M., Kwiatkowski, D.,1994. Variation in the TNF-a promoter region associated with susceptibility to cerebral malaria. Nature 371 : 508 – 511.
Messer,G.,Kick,G., Ranki,A., Koskimies,S., Reunala,T., Meurer,M., 1994. Polymorphism of the tumor necrosis factor genes in
patients with dermatitis herpetiformis. Dermatology 189 (Suppl. 1) : 135 – 137.
Micklos,D.A.,Freyer,G.A., Crotty,D.A.,1989.A Laboratory introduction to DNA restriction analysis.Biotechnology Education 1(1):16 - 22 Molvig, J.,Baek, L., Christensen,P., Manogue,K.R., Viassara,H., Piatz, P.,1988. Endotoxin-stimulated human monocyte
secretion of interleukin 1, tumor necrosis factor alpha, and prostaglandin E2 shows stable interindividual differences.
Moeslichan, S.,1990.Penelitian sistem HLA Dalam Upaya memperoleh sumber antibodinya.Disertasi.Universitas Indonesia.Jakarta
Molfino, N.A., 2004. Genetics of COPD. Chest 125,5: 1929 - 1940 Munir, D., 2007.Alel gen HLA DRB1 dan HLA DQB1 yang
berhubungandengankerentanantimbulnyaKarsinoma
NasofaringpadasukuBatak.Disertasi.Universitas Sumatera
Utara.Medan
Nedwin,G.E.,Naylor,S.L.,Sakaguchi,A.Y.,Smith,D.,Jarret-Nedwin, J., Pennica,D., Goeddel,D.V., GrayP.W.,1985. Human
lymphotoxin and tumor necrosis factorgenes: structure, homology and chromosomal localization.Nuclei Acids Res
11;13(17); 6361 - 6373
Ngestiningsih, D., danFaradz, S.M.H.,2005.Polimorfismealel TNFα
padapenderita DHF di Semarang. LaporanKegiatan
PenelitianIlmuPengetahuanDasar. FakultasKedokteran
UniversitasDiponegoro. Semarang
Nuswantara, S., Wulandari, A.P., Novel,S.S.,2010.Kamus Saku Biologi Molekuler. TIM.Jakarta
Opal S.M., and DePalo V.A.,2000. Anti Inflammatory Cytokines. Chest
117:1162-1172
Ozhan, G., Yanar, H.T., Ertekin, C., Alpertunga, B., 2010.
Polymorphisms in Tumor Necrosis Factor Alpha (TNFα)
Gene in Patients with Acute Pancreatitis.Mediators of Inflammation vol.2010,articleID482950, 6 pages
Patuzzo C., Gile,L.S., Zorzetto,M.,Trabetti,E.,Malerba,G.,Pignatti,P.F., Luisetti,M., 2000.Tumor Necrosis Factor Gene Complex in COPD and Disseminated Bronchiectasis.Chest2000;117
:1353-1358
PerhimpunanDokterParu Indonesia (PDPI)., 2011. PPOK (Penyakit ParuObstruktifKronik). Pedomanpraktis diagnosis dan
penatalaksanaan di Indonesia. Jakarta,PDPI.
PerhimpunanDokterParu Indonesia (PDPI)., 2003. PPOK (Penyakit\ ParuObstruktifKronik). Pedomanpraktis diagnosis dan
Petrescu, F., Voican S.C., and Silosi I., 2010. Tumor Necrosis Factor Alpha Serum Levels in Healthy Smokers and NonSmokers.
International Journal of Chronic Obstructive Pulmonary Disease 5 :217-222
Pociot, F., D’Alfonso, S., Compasso,S., Scorza, R., Richiardi,P.M., 1995. Functional analysis of a new polymorphism in the
human TNF a gene promoter. Scand J Immunol 42 : 501 – 504.
Purba M.Y., Pandia,P.S., dan Tarigan A.P., 2011.Profil Penderita Yang Didiagnosis Sebagai Penyakit Paru Obstruksi Kronik Di RSUP Adam Malik, RSU Tembakau Deli Dan RSU Pirngadi
Medan Tahun 2010. Tesis, Program Megister Kedokteran
Klinik Departemen Pulmonologi Dan Kedokteran Respirasi Fakultas Kedokteran USU/SMF Paru RSUP H.Adam Malik Medan.
Raherison, C., and Girodet, P.O.,2009. Epidemiology of COPD.Eur Respir Rev18114 ; 213 - 221.
Rasmin, M., 2008.KedokteranRespirasi, Pemahamansebuah
perjalanan.Pidatopadaupacarapengukuhansebagai
guru besartetapdalambidangPulmonologidanilmu
KedokteranRespirasipadaFakultasKedokteran,Universitas Indonesia.Jakarta.
Regional COPD Working Group., 2003. COPD prevalencein 12Asia Pacific countries and regions:projections based on the COPD prevalence estimation model. Respirology8 : 192 – 8.
Rennard, S., and Barnes, P.J., 2002.Pathogenesis of COPD. In: Barnes, P.J., et al., eds. Asthma and COPD. Basic
mechanisms and clinical management. Academic Press : 361 – 383.
Saiki,R.K.,1989. The design and optimization of the PCR.In PCR Tehnology:Principleand Application for DNA Amplification. H.A.Erlich, ed:7-16
Sakao, S., Tatsumi, K., Igari,H., Shino,Y., Shirasawa,H., Kuriyama,T.,
2001. Association of tumor necrosis factor α gene promoter
polymorphism with the presence of Chronic Obstructive Pulmonary Disease.Am J RespirCrit Care Med 163:420 – 422 .
Sanford, A.J., Chagani,T., Weir,T.D., Connet,J.E., Anthonisen,N.R., Pare,P.D.,2001. Susceptibility genes for rapid decline of lung function in the lung health Study. Am J RespirCrit Care163 :
469 – 473 .
Sapey, E., Wood,A.M., Ahmad,A., Stockley,R.A., 2010.Tumor Necrosis Factor–a rs361525 PolymorphismIs Associated with
Increased Local Productionand Downstream Inflammation in ChronicObstructive Pulmonary Disease.Am J Respir Crit
Care Med 182: 192–199
Sariban, E., Imamura, K., Luebbers, R., Kufe, D., 1988. Transcriptional and posttranscriptional regulation of tumor necrosis factor
gene expression inhuman monocytes. J. Clin. Invest. 81 : 1506 – 1510.
Siafakas, N., Tzortzaki, E., and Bouros, D., 2003. Why do only some smokers develop COPD? In : Pearson, M., Weidzcha, W.,
eds. Chronic Obstructive Pulmonary Disease : Critical Debates.Blackwell Science.
Silverman, E.K., 2002. Genetic epidemiology of COPD.Chest121 :1S- 6S
Smoolonska,J., Wijmenga,C., Postma,D.S., Boezen,H.M.,2009.Meta analyses on suspected chronicobstructive pulmonary
disease genes. A summary of 20 years' research. American Journal of Respiratory and CriticalCare Medicine180: 618 – 631.
Subowo,2009. Sitokindalamsistemimun.Dalam :
Subowo,Imunobiologi. Bandung, SagungSeto : 121 – 146. Susanto, A.D., 2009. Masalahrokokdanstrategiberhentimerokok. Dalam:Alvin Kosasi, NeniSawitri. Harmony of pulmonology update. Ilmupenyakitparudanaplikasinyauntukdokter
umum. Bogor, PanitiaPertemuanIlmiahParuBogor :119- 133.
Susenas, Survei Sosial Ekonomi Nasional.2001
Supardi I., dan Iswari, IS. Pengantar:Kedokteran Molekuler. Program Pascasarjana Combined Degree Fakultas Kedokteran
Universitas Padjajaran: 148-149
Pare, P.D., 2007. Tumor necrosis factor and lymphotoxin A polymorphisms and lung function in smoker.
EurRespir J29 : 31 – 41 .
Teramoto, S.,and Ishii, T., 2001. No association of tumor necrosis
factor gene α polimorphism and COPD in caucasion smokers andjapanesesmokers.Chest 119 : 315 – 316 .
Teramoto, S., 2007. COPD Pathogenesis from the viewpoint of risk factors. Internal Medicine 46 (2): 77 – 80 .
Turyadi, 2006. Frekuensi alel TNF2 dan Alel LTα pada penderita
malaria berat dan malaria ringan di Indonesia. Skripsi Srajana Sains Dalam Bidang Biologi.Fakultas Biologi Universitas Nasional.
Yuwono, T., 2005.Replikasi bahangenetik.Dalam :Yuwono, T. Biologi molekular. LaboratoriumMikrobiologi, FakultasPertanian,
UniversitasGajahmada: 94 – 132 .
Udalova, I.A., Richardson, A., Denys, A., Smith, C., Ackerman, H., Foxwell,B., 2000. Functional consequences of a
polymorphism affecting NFkB p50-p50 binding to the TNF promoter region. Mol Cell Biol20 : 9113 – 9119.
United Nations EnviromentProgramme. Globalenvironmental outlook, 2000. Availablefrom:URL
Van der vaart H.,Koeter,G.H., Postma, D.S., Kauffman,H.F., ten Hacken,N.H.,2005. First Study of infliximab treatment in
patients with chronic obstructive pulmonary disease.Am J Respir Crit Care Med 172: 465-469
Viegi, G., Pistelli, F., Sherrill,D.L., Maio,S., Baldacci,S., Carrozzi,L., 2007. Definition epidemiology and natural history of COPD.
EurRespir J30 : 993 – 1013.
Vestbo, J., and Lange, P.,2003 Epidemiology. In: Chronic Obstructive Pulmonary Disease. Arnold, second edition, London
Walter, R.E., Wilk,J.B., Larson, M.G., Vasan, R.S., Keaney,J.F., Lipinska, I., 2008. Systemic inflammation in COPD. The Framinghamheart study. Chest 133 : 19 – 25 .
Wan, E.S., and Silverman, E.K., 2009. Genetics of COPD and Emphysema.Chest 136 (3) :859 – 866.
WHO.Indoor air pollutan and household energy. Available from : URL ;
[Accessed on 5 July 2010]
WHO Report on the Global Tobacco Epidemic, 2008. Available from:
forward summary2008 .pdf. [Accessed on 5 July 2010]
Willemse,B.W., Postma,D.S., Timens, W., ten Hacken,N.H.,2004. The impact of smoking on respiratory symptoms, lung function,
airway hyperresponsiveness and inflammation. Eur Respir J 23:464-476.
Wilson, A. G., di Giovine, F.S., Blakemore,A.I., Duff, G.W.,1992. Single base polymorphism in the human tumor necrosis factor alpha gene detectable by NcoI restriction of PCR product.
Hum. Mol .Genet.1 : 353
Wilson, A. G., de Vries, N., van de Putte, L.B., Duff, G.W.,1995. A tumor necrosis factor alpha polymorphism is not associated with rheumatoid arthritis.Ann. Rheum. Dis. 54 : 601 – 603.
Wilson, A.G., Symons, J.A., McDowell,T.L., McDevitt,H.O., Duff,G.W., 1997. Effects of a polymorphism in the human tumor necrosis
factor a promoter on transcriptional activation. ProcNatlAcad Sci USA 94: 3195 – 3199.
Wilson,A.G., de Vries N., Pociot, P., di Giovine,F.S., van der Putte, L.B., Duff, G.W.,1993.An Allelic polymorphism withinthe
human tumor necrosis factor α promoter region is strongly
associated with HLA A1, B8 and DR3 alleles.J.Exp.Med. 177:557-560
Wiyono, H.W., 2009. PenyakitParuObstruksiKronik.Tantangandan
peluang.Pidatopadaupacarapengukuhansebagai guru
besartetapdalambidangpulmonologidanilmukedokteran respirasipadafakultaskedokteranUniversitas Indonesia, Jakarta.
Stockley, R.A., 2008. The TNFα gene relates to phenotype in 1-antitrypsin deficiency.RespirRes 9 : 52
WHO., 2009. Global tuberculosis control : epidemiology, strategy, financing. WHO report .WHO / HTM / TB / 2009.411 ,
Geneva, Switzerland
WHO., Annual event. Available from : http : // www. who. Int / mediacentre / events / annual / en /. [Accesed on 5 July2010] Wright, J.I., Tai, H., Wang, X., Churg, A., 2007. Cigarette smoke upregulatespulmonary vascular matrix metalloproteinases
via TNFα signaling. Am J Physiol Lung Cell MolPhysiol292 : L125 – L133.
Wu, W.S., and McClain, K.L., 1997.DNA polymorphisms and mutations of the tumor necrosis factor-a (TNF-a) promoter in
Langerhans cell histiocytosis (LCH). J Interferon Cytokine Res 17 : 631 – 635.
Zanobetti, A., Bind, M.A., and Schwartz, J., 2008. Particulate air polution and survival in COPD. Environmental Health 7 : 48
Zhai, G., Valdes, A.M., Cherkas,L., Clement, G., Strachan,D., Spector, T.D.,2007. The interaction of genes and Smokingon Forced Expiratory Volume* A Classic Twin Study.Chest 132 : 1772
– 1777.
Zhang C., Cai,S., Chen,P., Chen,J.B., Wu,S.J., Zhou, R., 2008. Inhibition of Tumor Necrosis Factor Alpha Reduce Alveolar SeptalCell Apoptosis in Passive Smoking Rats.Chin Med J, 121(7):597-601
Zhang,X.Y., Zhang,C., Sun,.Q.Y., Li,D., Luo,R.R., Wan, Z.F., 2011. Infiximab protects against pulmonary emphysema in smoking rats. Chin Med J 124(16):2502-6
Zhan, P., Wang, J., Wei, S.Z.,Qian, Q.,Qiu, L.X., Yu, L.K., 2011. TNF-308 gene polymorphism is associated with COPD risk among Asians: meta-analysis of data for 6,118 subjects. Mol Biol Rep 38:219-227
Hasil PCR -238TNF-α sampel 1-77 (expected band:150 bp)
Hasil PCR -238TNF-α sampel 78-125 (expected band:150 bp)
M M M M -C M M -C -C
Hasil PCR -238TNF-α sampel 126-152 (expected band:150bp)
Hasil PCR -238TNF-α sampel 153-190 (expectedband:150 bp)
Hasil PCR -238TNF-α sampel 191-227 (expected band:150 bp) M M M -C M M -C
Hasil RFLP -238TNF sampel 1-23
Hasil RFLP -238TNF sampel 24-69, sampel mutan (25,26,33,37,39)
M uC uC M M uC uC uC uC 25 26 33 37 39
Hasil RFLP-238TNF sampel 70-77, 153-190; sampel mutan (71)
Hasil RFLP -238TNF sampel 191-227, sampel mutan (194)
Gambar berikutnya untuk RFLP -238 TNF sampel 78-152 (acak) M M uC uC uC uC 194 uC 170 M uC uC 168 169 171 172 173 174175 176 177 178 179 180181 182 183 184185 186 187 188 189 190 M uC 71 70 72 73 77 74 75 76 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167
Hasil RFLP -238TNF, sampel mutan (96, 98, 112, 148, 135, 136, 83) Hasil -238 TNF RFLP 80 89 122 132 138
M
uC
λ
M M M uC uC uC uC uC uC 112 136 135 115 83 98 96 151 86 150 101 128 102 133 8410778 97 12413087 116 118 111 92 108149114 94 144127 140121 126 100 134 142145131 148 106 81 152 91 104 10995 103 144 99 113 117 143 110 147 146 82 85 109 79 119 120 130 86 129 137 88 125 93KETERANGAN
M = DNA marker
-C = Kontrol negatif pada proses PCR
uC = Uncut control = kontrol DNA yang tidak terpotong pada proses RFLP
λ = Lambda DNA sebagai kontrol reaksi RFLP
SIMPULAN
Hasil PCR semua sampel menunjukkan hasil yang cukup baik yaitu diperoleh DNA target TNF-α yang berukuran ±150 bp. Kontrol negatif muncul pada proses PCR kemungkinan disebabkan kontaminasi saat PCR, namun tidak mengganggu proses RFLP selanjutnya.
Sampel mutan dapat dibedakan dari sampel normal dengan melihat perbedaan ukuran pita DNA yang diperoleh pada hasil RFLP. Sampel mutan merupakan sampel yang menunjukkan pita yang panjangnya sama dengan kontrol DNA yang tidak terpotong. Total sampel mutan yang terdeteksi yaitu sebanyak 14 sampel mutan (25,26,33,37,39,71,96,98,112,115,135,136,152,194).
Hasil PCR -308TNF-α sampel 1-81 (expected band:230 bp) Tampak pita diatas 230 bp adalah smear yang muncul akibat taq polymerase yang kurang baik
-C -C
Hasil PCR -308 TNF-α sampel 163-227 (expected band:230 bp) -C
Hasil RFLP -308TNF sampel 1-48, sampel mutan (4,6,19,23,36,39,42)
Hasil RFLP -308TNF sampel 49-90, sampel mutan (49,63,66,67)
19 23 M C 4 6 36 39 42 19 23 M C 4 6 36 39 42 19 23 M C 49 63 66 67 M C 49 63 66 67
Hasil RFLP -308 TNF sampel 91-138, sampel mutan (118,119,129,130)
Hasil RFLP -308TNF sampel 139-180, sampel mutan (148,162) M C
118 119 129 130
M C 148 162
M C 148 162
Hasil RFLP -308TNF sampel 181-227 C C M M 205 212 219 201