CAJ | 학술논문

为了建立菜豆晕疫病菌的环介导等温扩增方法,本试验利用菜豆晕疫病菌argK特异性序列设计环介导等温核酸扩增( LAMP)引物,进行反应条件和反应体系的优化,并进行特异性和灵敏度验证.特异性检测结果显示,5株菜豆晕疫病菌的LAMP产物呈阳性结果,而参试的其他病原菌不产生扩增反应. LAMP检测基因组DNA和菌悬液时,其灵敏度分别达到0.632×10-4 ng/μ1和7.3×102 CFU/m1,该方法比常规PCR灵敏度高100倍.表明本研究所建立的LAMP检测方法简便、快速、准确、灵敏,可有效应用于口岸菜豆晕疫病菌的检测.
위료건립채두훈역병균적배개도등온확증방법,본시험이용채두훈역병균argK특이성서렬설계배개도등온핵산확증( LAMP)인물,진행반응조건화반응체계적우화,병진행특이성화령민도험증.특이성검측결과현시,5주채두훈역병균적LAMP산물정양성결과,이삼시적기타병원균불산생확증반응. LAMP검측기인조DNA화균현액시,기령민도분별체도0.632×10-4 ng/μ1화7.3×102 CFU/m1,해방법비상규PCR령민도고100배.표명본연구소건립적LAMP검측방법간편、쾌속、준학、령민,가유효응용우구안채두훈역병균적검측.
To deve1op a 1oop-mediated isotherma1 amp1ification ( LAMP ) method for detecting Pseudomonas savas-tanoi pv. phaseolicola, LAMP primers were designed based on the argK gene of P. savastanoi pv. phaseolicola, the reac-tion conditions were optimized, and the specificity and sensitivity of LAMP were tested. The amp1ification resu1ts revea1ed that five iso1ates of P. savastanoi pv. phaseolicola were positive, whi1e other pathogens were negative. LAMP method showed sensitivities of 6. 32×10-5 ng/μ1 and 7. 3×102 CFU/m1 when detecting genomic DNA and bacteria1 suspension of P. savastanoi, 100 times higher than conventiona1 PCR assay. The LAMP method deve1oped in this study was simp1e, fast, specific and sensitive, for detecting P. savastanoi pv. phaseolicola in port quarantine.