Polimeraz zincir reaksiyonu ve mikrobiyolojide kullanım alanları

İnfeksiyöz hastalıkların teşhisinde genellikle, direkt ve indirekt yöntemler kullanılır. İnfeksiyonların teşhisini sağlayan klasik direkt yöntemler (konvansiyonel teşhis yöntemleri) klinik bulgulara, otopsi bulgularına, etken izolasyon ve identifikasyonuna, antijenik materyallerin saptanmasına ve serolojik tekniklere dayanmaktadır. Bu yöntemler bazen başarılı sonuçlar vermediği gibi latent infeksiyonlarda da hatalı pozitif sonuçlara yol açabilmektedir. Bu gibi olgularda, mikroorganizmaya ait genetik materyallerin (DNA ya da RNA) veya proteinlerin saptanmasını amaçlayan, kesin sonuçlar veren biyoteknolojik teşhis yöntemlerinin kullanılmaları oldukça faydalı olmaktadır. Bu biyoteknolojik yöntemler içerisinde en yenilerinden birisi olan Polimeraz Zincir Reaksiyonu (PCR), spesifitesi ve sensitivitesinin oldukça yüksek olması gibi nedenlerle araştırmalarda yaygın olarak kullanılmaktadır.

Anahtar Kelimeler:

mikrobiyoloji, bulaşıcı hastalıklar, polimeraz zincir reaksiyonu, kullanım

Polymerase chain reaction and its usage in mirobiology

Direct and indirect methods are generally used in the diagnosis of infectious diseases. Classical direct (or conventional) diagnostic methods usually rely on clinical and post mortem findings, isolation and identification of the causative agents, detection of the antigenic substances and of specific antibodies by serologic methods. These tecniques may sometimes give insufficient results or may cause false positive results in latent infections. In such cases, the use of biotechnologic methods for the detection of microbial genetic materials (DNA or RNA) or proteins would be benifical. Polymerase Chain Reaction (PCR), one of the newest of these biotechnologic methods, is used widely in research studies because of its high sensitivity, specificity, reliability and accuracy.

Keywords:

microbiology, infectious diseases, polymerase chain reaction, usage,

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1.Abigail A, Salyers D, Whitt D: Bacterial Pathogenesis: A Molecular Approach, ASM Press, Washington DC, 1994.
2.Arda M: Biyoteknoloji (Bazı Temel İlkeler), Kükem Derneği Bilimsel Yayınları, Ankara, 1995.
3.Çevik AM: PCR ve İnfeksiyöz Hastalıklarda Kullanımı. Seminer, S.B. Ankara Hastanesi Klinik Mikrobiyoloji ve İnfeksiyon Hastalıkları Bölümü, 1994.
4.Chen X, Miflin JK, Zhang P, Blackall PJ: Development and application of DNA probes and PCR tests for Haemopilus paragallinarum. Avian Dis, 40: 398-407, 1996.
5.Diallo IO, Mackenzie AM, Spradbrow PB, Robinson WF: Field isolates of fowl pox virus contaminated with reticuloendotheliosis virus. Avian Dis, 27:60-66, 1998.
6.Diker KS: İmmünoloji. Medisan Yayınları Serisi, 37. Ankara, 1998.
7.Erlich AH, Gelfand D, Sninsky JJ: Recent advances in Polymerase Chain Reaction. Science, 252: 1643-1652, 1991.
8.Giarcia M, Jackwood WM, Levisohn S, Kleven HS: Detection of Mycopllasma gallisepticum, M. synoviae, and M. iowae by multispecies Polymerase Chain Reaction and restriction fragment length polymorphisim. Avian Dis, 39: 606-616, 1995.
9.Hungjen L, Giamborne JJ, Nielsen B, Liu H: Molecular characterization of avian reoviruses using nested PCR and nucleotid sequence analysis. J Virol Methods, 65:157-167, 1997.
10.Jestin V, Jestin A: Detection of Newcastle disease virus RNA in infected allantoic fluilds by in vitro enzymatic amplification ( PCR). Arch Virol, 118 : 151-161, 1991.
11.Kwok S, Higuchi R: Avoiding false positives with PCR. Nature, 339:237-38, 1989.
12.Kwon MH, Jackwood MW: Polymerase Chain Reaction and biotin-labeled DNA probe for detection of Infectious Bronchitis virus in chickens. Avian Dis, 37:149-156, 1992.
13.Miwa N, Nishina T, Kubo S, Honda H: Most probable number method combined with nested PCR for detection and enumerailon of -enterotoxigenic Clostridium perfiringens intestinal contents of tattle, pig and chicken. J Vet Med Sci, 59: 557-560, 1997.
14.Oyofa BA, Abd-el-salam M, Churilla M: Rapid and sensitive detection of Campylobaeterspp. from chicken using the Polymerase Chain Reaction. Zbl Bakt, 285:480-485, 1997.
15.Persing HD: Polymerase Chain Reaction: Trenches to benches. J Clin Microbiol, 29: 1281-1285, 1991.
16.Rodriguez JM: Detection of pathogens by using the Polymerase Chain Reaction. VetJ, 153: 287-305, 1997.
17.Saiki KR, Gelfand HD, Stoffi S, Scharf JS, Higuchi R, Horn TG: Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science, 1988.
18.Schochetman G, Jones KW: Polymerase Chain Reaction. J InfDIs, 158: 1154-1157,1988.
19.Sonie C, Watson k, Rybicki E, Luicio B: Determination of the detection limit of the Polymerase Chain Reaction for chicken Infectious Anemia virus. Avian Dis, 37: 467-476, 1993.
20. Starm YM, Meir R, Molad T, Blumenkranz R, Malkinson M: Application of the Polymerase chain Reaction detect Infectious Bursal Disease virus naturally infected chickens. Avian Dis, 38:879-884, 1994.
21.Tindall RK, Kunkel AT: Fidelity of DNA synthesis by the Thermus aquaticus DNA Polymerase. Biochemistry, 27: 6008-60013,1988.
22.Tunchili LM, Kodama H, Sharma RN, Takatori I, Pandey GS: Detection of Salmonella DNA in chicken embriyos and environmental samples by Polymerase Chain Reaction. J Vet Med Sci, 58: 881-884, 1996.
23.Walker J, Douan G: DNA probes: A new role in diagnostic microbilology. J Appl Microbiol, 67 : 229-230, 1989.
24.Wolcott JM: DNA - based rapid methods for the detection of foodborne pathogensJ. Food Protec, 54:387-401,1991.
25.Wolcott JM: Advances in nucleic asid - based detection methods. Clin Microbil Rev, 5 : 370-386, 1992.