Polymerase Chain Reaction-Restriction Fragment Length Polymorphism, A Rapid and Simple Method for Identification of Pathogenic Fungi

In recent years, the incidence of infections caused by pathogenic fungi has increased globally, especially in immunocompromised patients, transplant patients, or patients undergoing chemotherapy in intensive care units. Due to the morbidity and rapid spread of infection to other organs, accurate, rapid diagnosis and treatment of fungal infections is essential. Molecular methods such as polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) have significant superiority in sensitivity and specificity compared to conventional methods. The aim of this study is to point out the limitations of common diagnostic methods and factors affecting the sensitivity and specificity of PCR-RFLP molecular diagnostic test.
This article is a review study for which subject-related academic papers (before 2023) were collected and studied by searching the keywords of pathogenic fungi, and RFLP molecular diagnostic methods; in Persian databases as well as Latin electronic databases including Scopus, Web of Science, and PubMed .
Reviewing numerous articles showed that PCR-RFLP is a rapid, practical, and reliable method, which can be used in laboratories to determine the genotype of specific variants of fungal species isolated from clinical samples. However, there are several limitations for PCR-RFLP method demanding for more attention. The main limitations are the need for specific restriction enzymes and the difficulty of accurately determining single nucleotide polymorphisms (SNPs) in a specific diagnostic locus.