Establishment of IMSA-LAMP detection method for Ralstonia solanacearu

Objective  Mulberry bacterial wilt is a seriously harmful bacterial disease caused by Ralstonia solanacearum. Therefore, it is of great significance to establish a rapid and sensitive detection method for R. solanacearum to effectively control mulberry bacterial wilt.

Method  Pectate lyase gene of R. solanacearum was used as the target. Based on the primer design principle of isothermal multiple self-matching-initiated amplification (IMSA), combining the loop-mediated isothermal amplification (LAMP) reaction system, a rapid and effective IMSA-LAMP method for detection ofR. solanacearum was established. The optimal reaction parameters of this method were screened.

Result  The IMSA-LAMP method based on pectate lyase gene could complete the specific detection of positive samples within 45 min at 64.5 ℃, and the detection sensitivity of R. solanacearum template DNA was 200 fg/μL (the corresponding bacteria detection sensitivity was 1 × 10² CFU/mL); The detection rate of suspected mulberry bacterial wilt samples collected in production was 87.5%.

Conclusion  This method has good practicability and can provide new technical support for the rapid detection, diagnosis and epidemic prevention of mulberry bacterial wilt.