Internal amplification controls have not been employed in fungal PCR hence potential false negative results

R R M Paterson1

  • 1Centro de Engenharia Biológica, Micoteca da Universidade do Minho, Braga, Portugal. russell.paterson@deb.uminho.pt

Insights

Internal amplification controls (IACs) are crucial for accurate fungal polymerase chain reaction (PCR) testing. Their absence in most studies compromises results, necessitating their mandatory inclusion in future fungal PCR research for reliable disease and toxin detection.

Area of Science:

  • Mycology
  • Molecular Biology
  • Biotechnology

Background:

  • Fungi are vital in various industries (food, medicine) but challenging to identify.
  • Polymerase chain reaction (PCR) is increasingly used for fungal identification.
  • PCR can yield false negatives, misclassifying fungal samples as safe.

Purpose of the Study:

  • To highlight the critical need for internal amplification controls (IACs) in fungal PCR.
  • To assess the impact of IACs on the reliability of fungal detection methods.
  • To advocate for the mandatory implementation of IACs in future fungal research.

Main Methods:

  • Review of numerous scientific reports on fungal PCR applications.
  • Analysis of the prevalence and impact of internal amplification controls (IACs).
  • Identification of potential sources of inhibition in fungal cultures, such as secondary metabolites.

Main Results:

  • A significant lack of IAC implementation was observed across reviewed fungal PCR studies.
  • Only one study, dating back to 1995, utilized an IAC to address potential inhibition.
  • The absence of IACs compromises the accuracy and reliability of fungal detection results.

Conclusions:

  • Previous fungal PCR research is compromised due to the non-use of IACs.
  • Minimizing inhibitors in fungal cultures is essential for reliable PCR outcomes.
  • Future fungal PCR research must incorporate IACs from the outset to ensure data integrity.