SIGNAL AMPLIFICATION TECHNIQUES AND APPLICATIONS IN DIAGNOSTIC VIROLOGY

Year 2020, Volume: 83 Issue: 3, 293 - 301, 29.06.2020

Fatih Şahiner , Ramazan Gümral

Abstract

Signal amplification methods are the most preferred non-PCR molecular tools in diagnostic virology. The aim of signal amplification tests is to amplify and measure the signals obtained from the target DNA or RNA in clinical samples, instead of directly copying these molecules. With these characteristics, signal amplification methods do not pose any risk of contamination of the test samples, laboratory devices or working environment with amplified products. Approved signal amplification tests have been commercially available for detection and quantitation of HIV-1, HCV and HPV nucleic acids, as well as genotyping analysis of HPV. Signal amplification tests were first used in the early 1990s and have different working principles and special designs. These tests are now combined with new diagnostic techniques such as real-time PCR, the rolling circle method, luminex xMAP, DNA biosensor technology and sequence analysis, in the detection and genotyping of viral agents, multiplex analysis and the discovery of new and unknown viruses. In addition, approved signal amplification tests are used as the standard control methods for newly developed test designs. In this review, the working principles of signal amplification-based tests and their use in diagnostic virology are discussed in detail and examples of their use in other platforms are presented. 

Keywords

Branched DNA, Hybrid capture, Invader

SİNYAL AMPLİFİKASYON TEKNİKLERİ VE TANISAL VİROLOJİDEKİ UYGULAMALARI

Abstract

Tanısal virolojide PCR dışı moleküler araçlar arasında ilk sırada sinyal amplifikasyon yöntemleri yer almaktadır. Sinyal amplifikasyon testlerinde klinik örneklerdeki hedef DNA veya RNA’nın doğrudan kopyalanması yerine, bu moleküllerden elde edilen sinyallerin yükseltilmesi ve ölçülmesi amaçlanır. Bu özellikleri ile sinyal amplifikasyon yöntemleri incelenecek örnekler, kullanılan gereçler veya çalışma ortamının amplifiye ürünlerle kontamine olması bakımından risk taşımazlar. HIV-1, HCV ve HPV nükleik asitlerinin saptanması ve kantitasyonu ve HPV genotip analizi için onaylı sinyal amplifikasyon testleri ticari olarak kullanıma sunulmuştur. İlk olarak 1990’lı yılların başlarında kullanılmaya başlanan ve farklı çalışma prensipleri ve özel tasarımlara sahip olan sinyal amplifikasyon testleri günümüzde real-time PCR, rolling circle yöntemi, luminex xMAP, DNA biyosensör teknolojisi ve dizi analizi gibi yeni tanısal tekniklerle kombine edilerek viral etkenlerin saptanması, genotiplendirilmesi, multipleks analizleri ve bilinmeyen yeni virusların keşfedilmesinde kullanılmaktadır. Onaylı sinyal amplifikasyon testleri yeni geliştirilen test tasarımları için standart kontrol yöntemleri olarak da kullanılmaktadır. Bu derlemede sinyal amplifikasyon temelli testlerin çalışma prensipleri ve tanısal virolojideki kullanımları detaylı olarak ele alınmış ve diğer platformlardaki kullanımlarına dair örnekler sunulmuştur.

Keywords

Dallı DNA, Hibrid yakalama, İnvader

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