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Nucleic acid extraction from clinical specimens for PCR applications

Polymerase chain reaction (PCR) has been widely used due to its high specificity, sensitivity, and rapid turn-around time. However, the major limitation of PCR based diagnosis is inhibition of the reaction caused by a variety of components within clinical specimens. | Turk J Biol 30 (2006) 107-120 © TÜB‹TAK Review Article Nucleic Acid Extraction from Clinical Specimens for PCR Applications Ashabil AYGAN Department of Biology, Faculty of Art and Science, Çukurova University, Adana - TURKEY Received: 24.06.2005 Abstract: Polymerase chain reaction (PCR) has been widely used due to its high specificity, sensitivity, and rapid turn-around time. However, the major limitation of PCR based diagnosis is inhibition of the reaction caused by a variety of components within clinical specimens. The demand for PCR diagnosis in medical microbiology has highlighted the need for efficient methods of nucleic acid extraction. Although there has been progress in the simplification and purification of microbial nucleic acids, many researchers’ procedures are still inconvenient for routine use with any specimen and a universal method has not been devised yet. In this review, some of the basic principles for extraction procedures that have been used to date and the present situation of nucleic acid extraction are summarized. Key Words: PCR, Clinical Specimens, Inhibitors, DNA extraction Klinik Örneklerden PZR için Nükleik Asit Ekstraksiyonu Özet: Polymer zincir reaksiyonu (PZR), yüksek oranda özgüllü¤ü, hassasiyeti ve sonuçlar›n çabuk al›nmas›ndan dolay› yayg›n bir flekilde kullan›lmaktad›r. Fakat, PZR temelli teflhisin en büyük engeli klinik örneklerdeki çeflitli bileflenlerden kaynaklanan reaksiyonun inhibisyonudur. T›bbi mikrobiyolojideki PZR teflhisi için ra¤bet, etkili nükleik asit extraksiyonu ihtiyac› art›rm›flt›r. Mikrobiyal nükleik asit saflast›r›lmas› ve kolayl›¤› aç›s›ndan bir çok ilerleme yap›lm›fl olmas›na ra¤men, ço¤u metod hala bir çok örnek için günlük kullan›ma uygun olmay›p genel bir metod gelifltirilmemifltir. Bu çal›flmada bugüne kadar kullan›lan ekstraksiyon prosedürlerinin baz› temel prensibleri ve nükleik asit ekstraksiyonlar›n›n mevcut durumu k›saca özetlenmifltir. Anahtar Sözcükler: PCR, Klinik Örnekler, ‹nhibitörler, DNA