本文選題：益生菌　切入點：16S　出處：《第三軍醫(yī)大學(xué)》2006年碩士論文　論文類型：學(xué)位論文

【摘要】： 益生菌制劑產(chǎn)品菌種與活菌數(shù)含量是其產(chǎn)品質(zhì)量的關(guān)鍵。國外益生菌產(chǎn)品的微生物分析表明,部分產(chǎn)品的微生物種屬和活菌數(shù)含量與產(chǎn)品標(biāo)簽上標(biāo)注的種屬和數(shù)量不符。然而,國內(nèi)還沒有這方面的研究報道。本課題對國內(nèi)主要的五種醫(yī)用益生菌制劑產(chǎn)品進(jìn)行菌種分析�？疾炝宋宸N醫(yī)用益生菌制劑多個批次產(chǎn)品的活菌數(shù)含量,有統(tǒng)計意義的反映產(chǎn)品在有效期內(nèi)的活菌數(shù)含量情況。通過結(jié)合表型特征分析法與16S rDNA序列分析法對從產(chǎn)品中分離的10株菌進(jìn)行種屬鑒定。同時將PCR-DGGE分析方法應(yīng)用到鑒定益生菌制劑產(chǎn)品菌株組成,為益生菌制劑產(chǎn)品的質(zhì)量控制提供快速、可靠、重復(fù)性好的分析方法。 方法與結(jié)果:(1)通過篩選獲得到適合的選擇培養(yǎng)基,并用于5種醫(yī)用益生菌制劑25批產(chǎn)品的活菌計數(shù),計數(shù)結(jié)果顯示:產(chǎn)品Ⅰ中的三種菌的活菌數(shù)變化范圍分別為10~5～10~(10)CFU/粒、10~5～10~8CFU/粒、10~6～10~8 CFU/粒,產(chǎn)品Ⅱ的活菌數(shù)變化范圍為10~8～10~9 CFU/粒,產(chǎn)品Ⅲ的活菌數(shù)變化范圍為10~8～10~9 CFU/粒,產(chǎn)品Ⅳ中的兩種菌的活菌數(shù)變化范圍分別為10~7～10~8 CFU/粒和10~8～10~(10) CFU/粒,產(chǎn)品Ⅴ中的三種菌的活菌數(shù)分別為10~7 CFU/粒、10~6 CFU/粒、10~6 CFU/粒。(2)分離純化10株產(chǎn)品菌株,提取單菌株基因組DNA,PCR擴(kuò)增16S rRNA基因,純化并測定PCR產(chǎn)物序列,將測得序列與GenBank數(shù)據(jù)庫的序列進(jìn)行同源性分析,并建立系統(tǒng)進(jìn)化樹。在GenBank數(shù)據(jù)庫中登陸其10株菌的16S rDNA序列,獲得如下序列號:PFK1(DQ295034),PFK2(DQ295035),PFK3(DQ295036),LZCL(DQ295040),ZCS1(DQ295037),MMA1(DQ295041),MMA2(DQ295038),JSQ1(DQ295042),JSQ2(DQ295039),JSQ3(DQ295043)�；诰�16S rDNA序列的Blast和系統(tǒng)進(jìn)化樹分析結(jié)果表明產(chǎn)品Ⅰ中含有的三種菌分別為長雙歧桿菌、植物乳桿菌和糞腸球菌;產(chǎn)品Ⅱ中含有的菌是動物雙歧桿菌;產(chǎn)品Ⅲ中含有的菌是地衣芽孢桿菌;產(chǎn)品Ⅳ中含有的兩種菌是屎腸球菌和枯草芽孢桿菌;產(chǎn)品Ⅴ中含有的三種菌分別為動物雙歧桿菌、保加利亞乳桿菌、嗜熱鏈球菌。(3)將分離菌株進(jìn)行形態(tài)、生理和糖發(fā)酵實驗,結(jié)果表明這些表型鑒定的結(jié)論與16 S rRNA基因序列同源性分析結(jié)論一致,分離純化5種產(chǎn)品的10株菌中仍有相同的3株菌與標(biāo)簽標(biāo)注的不一致,即其中標(biāo)簽上分別標(biāo)注為青春雙歧桿菌和長雙歧桿菌的兩株菌鑒定結(jié)果均為動物雙歧桿菌,
[Abstract]:The microbiological analysis of probiotic products in foreign countries shows that the microbial species and the number of live bacteria in some products are not consistent with the species and quantity labeled on the product label. There is no such research report in China. In this paper, the main five kinds of medical probiotics products in China were analyzed, and the number of live bacteria in five kinds of medical probiotics products were investigated. It is statistically significant to reflect the content of live bacteria in the valid period of product. By combining phenotypic characteristic analysis and 16s rDNA sequence analysis, the species of 10 strains isolated from the product were identified. Meanwhile, the method of PCR-DGGE analysis was applied to identify the species of 10 strains isolated from the product. Used to identify the composition of probiotic product strains, To provide a rapid, reliable and reproducible analytical method for the quality control of probiotics. Methods and results: a suitable medium was obtained by screening and used to count live bacteria in 25 batches of 5 kinds of medical probiotics. The counting results showed that the number of live bacteria in product I ranged from 10 ~ 5 ~ 10 ~ (10) to 10 ~ (10) ~ 10 ~ (10) ~ (10) ~ (10) ~ (10) ~ (10) C ~ (10) C ~ (8) ~ (10) C ~ (10) C ~ (10) ~ (8) CFU/, to 10 ~ (8) ~ 10 ~ (9) CFU/ for product 鈪，

本文編號：1607690