【摘要】：【目的】建立一種快速、準(zhǔn)確的鼠李糖乳桿菌檢測(cè)方法。【方法】根據(jù)已報(bào)道的鼠李糖乳桿菌(Lactobacillus rhamnosus)的gyr B基因保守序列,設(shè)計(jì)用于檢測(cè)鼠李糖乳桿菌的特異性DPO引物,結(jié)合SYBR Green I實(shí)時(shí)熒光PCR技術(shù),建立鼠李糖乳桿菌的實(shí)時(shí)熒光PCR檢測(cè)方法,評(píng)價(jià)其特異性和靈敏度,并將建立的檢測(cè)方法用于實(shí)際樣品的檢測(cè)中�！窘Y(jié)果】該方法對(duì)2株鼠李糖乳桿菌能得到陽(yáng)性擴(kuò)增,而其余10種乳酸菌及陰性對(duì)照沒(méi)有擴(kuò)增曲線,而且在退火溫度為50~60℃不影響其特異性;該實(shí)時(shí)熒光PCR檢測(cè)靈敏度比農(nóng)業(yè)部標(biāo)準(zhǔn)中普通PCR檢測(cè)高10倍;用10種微生物肥料及其它益生菌產(chǎn)品進(jìn)行了驗(yàn)證,檢出情況與產(chǎn)品標(biāo)識(shí)一致�！窘Y(jié)論】研究建立的SYBR GreenⅠ實(shí)時(shí)熒光PCR能夠準(zhǔn)確、高效的檢測(cè)微生物肥料及其他益生菌產(chǎn)品中的鼠李糖乳桿菌。
[Abstract]:[objective] to establish a rapid and accurate method for the detection of Lactobacillus rhamnoides. [methods] based on the conserved sequence of gyr B gene of Lactobacillus rhamnoides (Lactobacillus rhamnosus), a specific DPO primer was designed for the detection of Lactobacillus rhamnoides. Combined with SYBR Green I real-time fluorescence PCR technique, a real-time fluorescence PCR method for detection of Lactobacillus rhamnoides was established, and its specificity and sensitivity were evaluated. The established method was applied to the detection of actual samples. [results] two strains of Lactobacillus rhamnoides were positive amplified by this method, while the other 10 lactic acid bacteria and negative controls had no amplification curve. Moreover, the specificity was not affected at the annealing temperature of 50 ~ 60 鈩，

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