AUTHOR=Hong-min Zhou , Jian Yu , Ying Liu , Yuan Yuan , Cui-ping Wu , Yu-cheng Dai , Jia-jia Chen 

TITLE=Rapid detection of Heterobasidion annosum using a loop-mediated isothermal amplification assay

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=13

YEAR=2023

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2023.1134921

DOI=10.3389/fcimb.2023.1134921

ISSN=2235-2988

ABSTRACT=<p><italic>Heterobasidion annosum</italic> is one of the most aggressive pathogens of <italic>Pinus</italic> forests in Europe, causing considerable economic losses. To detect <italic>H. annosum</italic> for disease diagnosis and control, we developed a loop-mediated isothermal amplification (LAMP) reaction with a primer set designed from the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) DNA sequences of <italic>H. annosum</italic>. In our study, this LAMP assay was found to be capable of efficiently amplifying the target gene within 60 min at 63°C. In specificity tests, <italic>H. annosum</italic> was positively detected, and other species were negative. The detection limit of this assay was found to be 100 pg·μL<sup>-1</sup>, and the assay was also successfully tested for use with basidiospore suspensions and wood samples. This study provides a rapid method for diagnosing root and butt rot caused by <italic>H. annosum</italic>, which will be of use in port surveillance of logs imported from Europe.</p>