Fiziksel ve Kimyasal Koşulların Çeşitli Yöntemlerle Elde Edilmiş Viral DNA’ya Etkileri

Year 2018, Volume: 7 Issue: 2, 149 - 153, 27.12.2018

Zeynep Akkutay Yoldar

https://doi.org/10.31196/huvfd.508964

Abstract

DNA virusları tamir mekanizmaları ve RNA viruslarına
göre daha stabil olmaları sayesinde çevre şartlarına dirençli bir yapı
gösterirler. Sahip oldukları bu direnç mutasyonel değişiklikleri elimine
ederek, virusun korunmuş bölgelerinin kolaylıkla amplifikasyonu sağlamasına
rağmen, uygun olmayan laboratuvar koşulları ve inhibe edici ajanlar, viral DNA
tespitine engel olabilmektedir. Viral DNA eldesinde çeşitli yöntemlerin
karşılaştırılmasının yapıldığı bu çalışmada aynı zamanda farklı fiziksel ve
kimyasal koşulların viral nükleik asite olan etkileri de sorgulandı. Bu amaçla
biri konvansiyonel metot olmak üzere toplam dört farklı ekstraksiyon yöntemi
bir parapoxvirus olan ecythma contagiosumun DNA’sını tespit amacıyla kullanıldı.
Elde edilen DNA’lar verimlilikleri yönünden kıyaslandıktan sonra çeşitli
fiziksel (basınç, UV) ve kimyasal (deterjan) koşullara maruz bırakıldı. Basınçlı
buhar otoklavına maruziyetten sonra viral DNA amplifikasyonu gerçekleşirken;
tween 20 ve uv radyasyon viral DNA amplifikasyonu inhibe ettiler. Ticari kitlerin
çoğu DNA ekstraksiyonunun süresini kısaltmaya yardımcı olurken konvansiyonel
yöntem viral DNA tespiti açısından en yüksek verimliliği sağladı.

Keywords

Ekstraksiyon, PCR, Viral DNA

The Effects of Physical and Chemical Conditions on Viral DNA Obtained by Various Methods

Abstract

DNA viruses are more resistant to environmental
conditions than RNA viruses due to their repair mechanisms and stablity of
their genetic material. Despite these advantages, inadequate laboratory
conditions and inhibitory agents can interfere with viral DNA detection. In
this study, which compares various methods of viral DNA obtainment, the effects
of different physical and chemical conditions on viral nucleic acid were also
questioned. For this purpose, a total of four different extraction methods, one
of which was the conventional method, have been used to detect the DNA of the
parapox virus causing ecythma contagiosum. The isolated DNAs were exposed to
various physical (pressure, uv) and chemical treatment (detergent) conditions before
PCR amplification. While viral DNA amplification occured after exposure to autoclave
pressure steam sterilizer; Tween 20 and UV radiation inhibited viral DNA amplification.
While most commercial kits helped to shorten the duration of DNA extraction,
the conventional method provided the highest efficiency for viral DNA
detection.

Keywords

Extraction, PCR, viral DNA

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