Published July 6, 2020 | Version v1
Journal article Open

PCR Inhibitors: Sources, Inhibition Mechanisms and Control

Creators

  • 1. Department of Medical Microbiology, Gulhane Training and Research Hospital, Ankara, Turkey

Description

Özet

Polimeraz Zincir Reaksiyonu (Polymerase Chain Reaction, PCR) temelli testlerin ilk örnekleri ve denemeleri üzerinden 40 yıla yakın bir zaman geçti. Bu süreçte PCR temelli moleküler testler gıda güvenliği, adli incelemeler, çevresel örneklerin incelenmesi, rekombinan gen teknolojisi, insan-hayvan-bitki-mikroorganizma genetiği ve immünoloji gibi alanlarda yaygın olarak kullanıldığı gibi en önemli kullanım alanlarından biri de enfeksiyon hastalıklarının tanısı ve takibi olmuştur. 2020 yılının ilk aylarından itibaren tüm dünyayı etkisi altına alan SARS-CoV-2 pandemisi ile mücadelede ülkelerin günlük test kapasiteleri ön plana çıkmış ve bu kapasitenin bazı ülkelerin salgın yönetimindeki başarısı üzerinde önemli etkileri olduğu üzerinde durulmuştur. Bu süreçte kısa bir zaman içerisinde çok sayıda kullanıcı tasarımlı (in-house) ve ticari SARS-CoV-2 RNA testi dizayn edilirken, PCR temelli tanı testleri belki de tarihte hiç olmadığı kadar yoğun bir şekilde kullanılmıştır. Türkiye’nin de dahil olduğu gelişmiş sağlık altyapısına sahip birçok ülkede bu testlerin kullanım onayı, tedarik ve pazarlama süreçleri sıkı denetimlere tabi tutulmuştur. Bu makalede çevresel (kanalizasyon suları, çevresel yüzeyler, hava örneklemeleri ve gıdalar) ve biyolojik örneklerde (hayvan örnekleri, hücre kültürleri, insanlardan alınan kan, solunum yolu, idrar, doku ve dışkı örnekleri gibi klinik örnekler) yapılan PCR testleri için önemli bir problem olan ve yanlış negatif sonuçların önemli bir nedeni olan inhibitör maddelere dikkat çekmek amaçlanmıştır. Makalede ayrıca inhibitör maddelerin olumsuz etkilerini ortadan kaldırmak için geliştirilen çeşitli önlemler ve inhibisyon kontrolünün önemi üzerinde durulmuştur.

Abstract

Nearly 40 years have passed since the first trials and prototypes of Polymerase Chain Reaction (PCR) based tests. In this period, PCR-based molecular tests have been widely used in the fields of food safety, forensic examinations, examination of environmental samples, recombinant gene technology, human-animal-plant-microorganism genetics, and immunology, as well as in diagnosis and monitoring of infectious diseases. In the fight against the SARS-CoV-2 pandemic, which has globally impacted the world since the first months of 2020, countries' daily testing capacities have gained importance and it was emphasized that this capacity had important effects on the success of some countries in epidemic management. In this process, while many in-house and commercial SARS-CoV-2 RNA tests were designed in a short time, PCR based diagnostic tests were used more extensively than ever before. The usage approval, procurement and marketing processes of these tests have been subjected to strict inspections in many countries, including Turkey, with advanced health infrastructure. In this article we aimed to draw attention to inhibitory substances that are a major cause of false negative results in PCR assays carried out in environmental (sewage waters, environmental surfaces, air samples and foods) and biological samples (clinical samples such as animal samples, cell cultures, blood samples from human, respiratory tract, urine, tissue and feces) in this article. The article also focused on various measures taken to eliminate the negative effects of inhibitory substances and the importance of inhibition control.

Notes

PCR İnhibitörleri: Kaynakları, İnhibisyon Mekanizmaları ve Kontrol

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