摘要
An improved electrochemiluminescence polymerase chain reaction （ECL-PCR） method was developed and applied to detect Fusarium wilt. Briefly, the internal transcribed spacer （ITS） sequence of Fusarium oxysporumf, sp Cubense （FOC） was amplified by PCR. Two universal fragments, which were complimentary to Ru（bpy）3^2＋ （TBR） labeled probe and Biotin labeled probe, respectively, were connected to the tail of primers so that all the PCR products got universal sequences. Then biotin labeled probes and TBR labeled probes were hybridized with the PCR products at the same time. Through the specific interaction between biotin and streptavidin, the PCR products were captured by streptavidin coated magnetic bead and then detected by ECL assay. The experiment results showed that the healthy banana samples and infected ones can be discriminated by this ECL-PCR method. This improved ECL-PCR approach is useful in Fusarium wilt detection due to its high sensitivity, simplicity and stability.
An improved electrochemiluminescence polymerase chain reaction （ECL-PCR） method was developed and applied to detect Fusarium wilt. Briefly, the internal transcribed spacer （ITS） sequence of Fusarium oxysporumf, sp Cubense （FOC） was amplified by PCR. Two universal fragments, which were complimentary to Ru（bpy）3^2＋ （TBR） labeled probe and Biotin labeled probe, respectively, were connected to the tail of primers so that all the PCR products got universal sequences. Then biotin labeled probes and TBR labeled probes were hybridized with the PCR products at the same time. Through the specific interaction between biotin and streptavidin, the PCR products were captured by streptavidin coated magnetic bead and then detected by ECL assay. The experiment results showed that the healthy banana samples and infected ones can be discriminated by this ECL-PCR method. This improved ECL-PCR approach is useful in Fusarium wilt detection due to its high sensitivity, simplicity and stability.
基金
the National Natural Science Foundation of China(No.3060012830470494)
the Natural Science Foundation of Guangdong Province(No.70058257117865).
