Powerful qPCR assays for the early detection of latent invaders: interdisciplinary approaches in clinical cancer research and plant pathology

Abstract

Latent invaders represent the first step of disease before symptoms occur in the host. Based on recent findings, tumors are considered to be ecosystems in which cancer cells act as invasive species that interact with the native host cell species. Analogously, in plants latent fungal pathogens coevolve within symptomless host tissues. For these reasons, similar detection approaches can be used for an early diagnosis of the invasion process in both plants and humans to prevent or reduce the spread of the disease. Molecular tools based on the evaluation of nucleic acids have been developed for the specific, rapid, and early detection of human diseases. During the last decades, these techniques to assess and quantify the proliferation of latent invaders in host cells have been transferred from the medical field to different areas of scientific research, such as plant pathology. An improvement in molecular biology protocols (especially referring to qPCR assays) specifically designed and optimized for detection in host plants is therefore advisable. This work is a cross-disciplinary review discussing the use of a methodological approach that is employed within both medical and plant sciences. It provides an overview of the principal qPCR tools for the detection of latent invaders, focusing on comparisons between clinical cancer research and plant pathology, and recent advances in the early detection of latent invaders to improve prevention and control strategies.

Fast and reliable molecular methods to detect fungal pathogens in woody plants

“…For the detection of pathogenic fungi in asymptomatic woody tissues, an appropriate sampling technique should be considered and associated with the detection protocol, as it will largely determine its sensitivity. The qPCR represents the gold standard for the quantitative measurement of nucleic acids and is thus the more appropriate tool for the early detection and quantification of latent invaders (Luchi et al 2016(Luchi et al , 2020. The sensitivity of this approach has been developed to detect low amount of the DNA of fungal pathogens, which could become harmful to trees stressed by abiotic factors as water deficiency (Luchi et al 2005a(Luchi et al , 2005b(Luchi et al , 2006.…”

“…In the future, it seems equally important to have tools and, above all, a sampling method to obtain data regarding the prevalence of host plants that are infected in a cryptic manner and therefore asymptomatic but potentially infectious (Luchi et al 2016;Thompson et al 2016). As already demonstrated for F. circinatum (Storer et al 1998;Coutinho et al 2007), infection with many phytopathogenic fungi may remain cryptic until environmental conditions or stress induce the switch to the pathogenic form (Agrios 2005).…”

New insight on DNA isolation from single automated recoveredCeratocystis plataniconidia

“…Most molecular techniques have been transferred from the clinical field to the field of plant pathology. The sensitivity of these approaches allows pathogens to be detected even before symptoms had developed on the plant [17][18]. The use of these new approaches based on single-cell analysis may also be relevant to study cell biology in plant-pathogenic microorganism such as fungi and bacteria.…”

Detection and quantification of <i>Colletotrichum abscissum</i> from leaves of budwood increase block and citrus nursery plants by real time PCR

“…To overcome this problem, molecular techiques such as real-time or quantitative polymerase chain reaction (qPCR) can be useful in this task. The qPCR is currently considered the most efficient technique for detection of phytopathogens, since it allows quantifying the amount of target nucleic acid in the sample, with higher assay sensitivity and specificity than conventional detection methods (CAPOTE et al, 2012;LUCHI et al, 2016). This technique allows monitoring of the reaction during the amplification process by using a fluorescence signal, which increases proportionally with the number of amplicons produced and the number of targets present in the sample (WITTWER et al, 1997;KURIAN et al, 1999;COCKERILL and SMITH, 2002).…”