Internal amplification controls have not been employed in fungal PCR hence potential false negative results.
Polymerase chain reaction (PCR) is subject to false negative results. Samples of fungi with the genes of interest (e.g. a disease or mycotoxin) may be categorized as negative and safe as a consequence. Fungi are eukaryotic organisms that are involved in many fields of human activity such as antibiotic, toxin and food production. Certain taxa are implicated in human, animal and plant diseases. However, fungi are difficult to identify and PCR techniques have been proposed increasingly for this purpose. Internal amplification controls (IACs) will ameliorate the situation and need to become mandatory. These are nucleic acids that posses a sequence which will provide a PCR product (i) using the same primers employed for the target gene, and (ii) that will not coincide on the gel with the product of the target gene. Only one group of workers employed an IAC, to respond to potential inhibition, which was reported in 1995 from this present assessment of numerous reports. Inhibitors in cultures need to be minimized, and secondary metabolites are an obvious source. The fields reviewed herein include medical mycology, mycotoxicology, environmental mycology and plant mycology. The conclusion is that previous reports are compromised because IACs have not been employed in fungal PCR; future research must include this control at an early stage.
Paterson
R R M
RR
Centro de Engenharia Biológica, Micoteca da Universidade do Minho, Braga, Portugal. russell.paterson@deb.uminho.pt
D000348:Aflatoxins / Q000235:genetics; D001230:Aspergillus / Q000235:genetics; D002273:Carcinogens / Q000032:analysis; D005188:False Negative Reactions; D005658:Fungi / Q000235:genetics*; D005670:Fusarium / Q000235:genetics; D005800:Genes, Fungal / Q000235:genetics*; D009793:Ochratoxins / Q000032:analysis; D010365:Patulin / Q000032:analysis; D010407:Penicillium / Q000235:genetics; D010838:Phytophthora / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*
D000348:Aflatoxins; D002273:Carcinogens; D009793:Ochratoxins; C025589:ochratoxin A; D010365:Patulin