本文選題：志賀菌 + 蘭州市區(qū)��； 參考：《蘭州大學(xué)》2008年碩士論文

【摘要】： 目的:調(diào)查蘭州市區(qū)腹瀉患者中志賀菌的感染率和感染型別;確定蘭州市區(qū)分離志賀菌株的毒力基因型;測定不同毒力基因型志賀菌的毒力,以探討毒力與毒力基因型的關(guān)系。 方法:選擇蘭州市區(qū)三家三甲醫(yī)院門、急診急性腹瀉患者,采用分離培養(yǎng)、生化反應(yīng)和血清學(xué)鑒別確定蘭州市區(qū)腹瀉患者中志賀菌的感染率和感染型別;設(shè)計引物,應(yīng)用聚合酶鏈反應(yīng)(PCR)方法,檢測志賀菌腸毒素1基因(setlA和setlB)、志賀菌腸毒素2基因(sen)、侵襲性質(zhì)粒抗原H基因(ipaH)、侵襲相關(guān)位點(diǎn)基因(ial)以及多重調(diào)控基因virA,確定蘭州市區(qū)分離志賀菌株的毒力基因型;以腹腔注射為感染途徑,測定不同毒力基因型的志賀菌對昆明種小鼠的半數(shù)致死量(LD50)。 結(jié)果: 1.從80例急性腹瀉患者糞便中分離出志賀菌16株,陽性率為20.0%。其中福氏志賀菌9株,占56.2%,宋內(nèi)志賀菌6株,占37.5%,痢疾志賀菌1株,占6.2%。同一菌群的不同志賀菌間在生化反應(yīng)和抗原性方面存在差異。 2.蘭州市區(qū)的志賀菌毒力基因virA、setlA陽性率最高,為100%;ipaH次之,陽性率為93.7%;ial、sen陽性率分別為56.2%、62.5%;setlB陽性率最低,僅為25.0%。除virA、ipaH、setlA外,宋內(nèi)志賀菌sen陽性率較高,為50.0%;ial檢出率較低,為33.3%,6株宋內(nèi)志賀菌均未檢出setlB。福氏志賀菌ial、setlB、sen陽性率分別為66.7%、44.4%、66.7%均高于宋內(nèi)志賀菌。 3.7株不同毒力基因型(virA+ipaH+setlA,virA+ial+ipaH+setlA+setlB+sen,virA+setlA,virA+ial+ipaH+setlA+sen,virA+ipaH+setlA+sen,virA+ipaH+setlA+setlB,virA+ial+ipaH+setlA+setlB)志賀菌LD50測定的結(jié)果分別為:1.422×10~9cfu/ml、6.653×10~7 cfu/ml、陰性、6.653×10~8 cfu/ml、1.422×10~9 cfu/ml、1.422×10~9cfu/ml、6.653×10~8 cfu/ml;對照宋內(nèi)志賀菌51334的LD50為3.062×10~9 cfu/ml。 結(jié)論: 1.蘭州市區(qū)腹瀉患者中志賀菌的感染率為20.0%左右,感染菌群以福氏志賀菌為主,其次為宋內(nèi)志賀菌。宋內(nèi)志賀菌感染的比例相對較高,占37.5%。 2.蘭州市區(qū)志賀菌中存在virA、ial、ipaH、setlA、setlB、sen毒力基因,以virA,setlA,ipaH較穩(wěn)定。志賀菌相關(guān)毒力基因檢測中,福氏志賀菌檢出率高于宋內(nèi)志賀菌。宋內(nèi)志賀菌攜帶setlA。 3蘭州市區(qū)志賀菌的毒力與毒力基因型密切相關(guān),以virA、ial、ipaH、setlA、setlB和sen均陽性的福氏志賀菌Ⅳ型毒力最強(qiáng)。 4.virA,setlA,ipaH基因可作為靶基因用于蘭州市區(qū)志賀菌的檢驗(yàn)。setlB可用于篩選福氏志賀菌Ⅳ型菌株。
[Abstract]:Objective: to investigate the infection rate and type of Shigella in diarrhea patients in Lanzhou city, determine the virulence genotype of Shigella isolated from Lanzhou city, and determine the virulence of Shigella virulent in different virulence genotypes, so as to explore the relationship between virulence and virulence genotypes. Methods: the isolated culture, biochemical reaction and serology were used to identify the infection rate and type of Shigella in three hospitals of Lanzhou district, and the primers were designed. Polymerase chain reaction (PCR) method was used. The virulence genotypes of Shigella enterotoxin 1 gene, Shigella enterotoxin 2 gene, invasive plasmid antigen H gene, invasion related locus gene, and multiple regulatory gene vira were detected, and the virulence genotypes of Shigella isolates were determined. The lethal dose (LD50) of Shigella with different virulence genotypes on Kunming mice was determined by intraperitoneal injection. Results: 1. Sixteen Shigella strains were isolated from the feces of 80 patients with acute diarrhea, and the positive rate was 20.0%. Among them, 9 strains of Shigella flexneri, 6 strains of Shigella sonnei, 1 strain of Shigella flexneri, 6 strains of Shigella sonnei, 1 strain of Shigella flexneri, accounted for 6. 2% of Shigella flexneri. There were differences in biochemical reaction and antigenicity among different Shigella strains of the same flora. 2. The positive rate of Shigella virulence gene virAtsetlA was the highest in Lanzhou city, with a positive rate of 100% ipaH, followed by 93.7% ialsen positive rate. The positive rate of Shigella virulence gene was 56.2% and 62.5% respectively, the lowest was 25.0%. The positive rate of sen in Shigella sonnei was higher than that in the other 6 strains of Shigella sonnei, and the detection rate of 50.0ial Shigella was lower than that of 33.3 strains of Shigella sonnei, and no setlB was detected in all the 6 strains of Shigella sonnei. The positive rates of Shigella flexneri were 66.7% and 44.4%, respectively, which were higher than those of Shigella sonnei. The results of LD50 determination of ial ipaH setlA setlB senvira ial ipaH setlA senvirA setlA ial setlA setlB of Shigella flexneri The LD50 of Shigella sonnei 51334 was 3.062 脳 10 9 cfuml-1 / ml, and the LD50 of Shigella sonnei 51334 was 3.062 脳 10 9 cfuml-1 路ml, and 1.422 脳 10 9 cfumlr / ml of 1.422 脳 10 9 cfuml-1 / ml, respectively, and the LD50 of Shigella sonnei 51334 was 3.062 脳 10 9 cfuml / r / ml, and the LD50 of Shigella sonnei 51334 was 3.062 脳 10 9 cfuml-1 / ml, instead of 1. 422 脳 10 9 cfuml / r = 1.422 脳 10 9 cfuml-1 / ml. Conclusion: 1. The infection rate of Shigella was about 20.0% in diarrhea patients in Lanzhou city. Shigella flexneri was the main infection group, followed by Shigella sonnei. The proportion of Shigella sonnei infection was relatively high, accounting for 37.5%. The virulence gene of Shigella spp. The detection rate of Shigella flexneri was higher than that of Shigella sonnei. The virulence of Shigella sonnei carrying setlA.3 was closely related to virulence genotypes. The virulence of Shigella flexneri type 鈪，

本文編號：2015273