The development and application of a LAMP assay for visualized detection of Chinese olive anthracnose

2020 
Anthracnose caused by Colletotrichum gloeosporioides (C. gloeosporioides) is an important disease affecting Chinese olive [Canarium album (Lour.) Rauesch] production. Therefore, developing a rapid and accurate method for C. gloeosporioides detection is of great importance for early diagnosis and immediate control of anthracnose. In the following study, we designed specific primers targeting the translation elongation factor alpha 1 gene of C. gloeosporioides, and established a loop-mediated isothermal amplification (LAMP) assay for fast and visualized detection system of C. gloeosporioides. The specificity and sensitivity of the LAMP assay were also measured. In the specificity test, characteristic bands were developed from 33 C. gloeosporioides isolates, and all of LAMP products showed green fluorescence after staining with SYBR Green I. On the other hand, negative results were obtained from C. gloeosporioides close species and other 19 common pathogens. The lower limit of LAMP detection was 0.4 fg·μL−1. This study provides a simple, highly sensitive, rapid and reliable technique for the detection of Chinese olive anthracnose.
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