Current approaches to development of real-time qPCR test-kits

Abstract

Currently fluorescent quantitative real-time polymerase chain reaction, which is a cutting-edge technology in genetic diagnosis, is used in different areas of molecular biology. Practical advantage of simplicity as well as combination of high speed, sensitivity and specificity made it possible to use this analysis for nucleic acid quantitation. The paper presents general information and recommended rules for the development of real-time qPCR. The publication is aimed to acquaint the researchers and reviewers with necessary requirements to be followed in order to ensure high accuracy, reliability and transparency of the experiments, correct interpretation and repeatability of the test results. Current approaches are described that allow obtaining reliable and consistent results by different operators, at different times and in different laboratories. Basic requirements for reagents used, nucleotide sequences and validation methods are given. In general, the publication gives the information needed to achieve three ultimate goals: to provide the authors with a broad range of tools and requirements for the development of real-time qPCR based-techniques; to give the possibility to the reviewers and editors of assessing the quality of articles and guidelines/instructions in accordance with the required criteria; to obtain consistent and reliable results of tests performed using this method.