PCRC: Avoiding Cross-Reactivity in Real-Time PCR Diagnostic Kits

BACKGROUND AND OBJECTIVESIntroduction: Diagnostic kits that utilize Real-Time PCR technology allow for identifying pathogenic microorganisms by detecting a portion of their genome. Specific primers and probes are designed for the target microorganism's genome to achieve this. During PCR, these primers amplify the target genome if present in the sample. The specificity of these primers and probes is crucial to avoid cross-reactivity and prevent the production of PCR products from other pathogens. When designing and producing diagnostic kits, in silico checks for potential cross-reactivity with other pathogens must be performed.MATERIALS AND METHODSIn this study, we developed a web-based tool called PCRC (Primer Cross-Reactivity Check) using Python programming language and bioinformatics algorithms. PCRC is used to determine the specificity of primers and probes designed to detect target microorganisms relative to the genomes of other pathogenic microorganisms.RESULTS AND DISCUSSIONIn this project, we were able to use the PCRC web application to review the results of probes and primers designed to identify pathogenic microorganisms and see the results indicating false positives.CONCLUSIONPCRC is a web-based program that performs cross-reaction tests by selecting the desired organisms, allowing for the elimination of false positive results in PCR tests.

کلیدواژه ها:

Real-Time PCR ، diagnostic kits ، primers ، probes ، cross-reactivity