Development of a point-of-care testing technology for pine wood nematode based on recombinase polymerase amplification
Through the analysis of the genome of Bursaphelenchus xylophilus and recombinase polymerase amplification(RPA)technology,a point-of-care testing product on pine wood nematode for field test was designed and produced.The results indicated that a specific gene,Bx-gene12,was identified through genome analysis of B.xylophilus.Based on this,specific primers and probes were designed.Grinding reagents were used to break nematode samples collected from pine trees in different regions.The deoxyribonucleic acids(DNA)of nematodes were effectively released by shaking at 1 500 revolutions per minute for 5 minutes.Under isothermal amplification at 40℃for 15 minutes,the detection sensitivity reached 100 fg or 1 nematode,effectively distinguishing B.xylophilus from other nematodes.Samples were taken from the wilted Pinus thunbergii and Pinus densiflora,the point-of-care testing method could effectively identify B.xylophilus in 25 minutes.This study utilized RPA technology to achieve point-of-care testing of B.xylophilus in the wild within 25 minutes,which can provide support for the timely and effective prevention and control of pine wilt disease.
Bursaphelenchus xylophiluspoint-of-care testingisothermal amplificationtechnical system
国家科技期刊平台
NSTL
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