幽門螺桿菌感染小鼠呼氣試驗(yàn)?zāi)Ｐ徒⒓捌溆绊懸蛩胤治?/H1>
發(fā)布時(shí)間：2018-11-23 14:47
【摘要】：目的：通過建立幽門螺桿菌小鼠13C-UBT、14C-UBT模型，探究構(gòu)建小鼠呼氣模型的條件和方法，分析影響幽門螺桿菌小鼠呼氣模型儀器、條件及生物因素，為研究幽門螺桿菌動(dòng)物模型提供一種無創(chuàng)性實(shí)時(shí)檢測(cè)方法，客觀評(píng)價(jià)HP疫苗在小鼠模型中的免疫保護(hù)效果以及對(duì)H.pylori致病性的深入研究提供一種簡(jiǎn)單、快速、便宜而且可靠的診斷手段。 方法： 利用6~8周齡BABL/c小鼠構(gòu)建小鼠呼氣模型的條件方法：通過對(duì)小鼠飼喂不同13C-Urea、14C-Urea藥物劑量，，間隔不同時(shí)間檢測(cè)，建立13C-UBT、14C-UBT實(shí)驗(yàn)?zāi)Ｐ�；飼喂不同H.pylori菌量，通過相同時(shí)間檢測(cè)，探究13C-UBT在相同檢測(cè)時(shí)間中飼喂不同菌量的小鼠的呼氣關(guān)系；小鼠飼喂不同H.pylori菌量，不同時(shí)間檢測(cè)，探究13C-UBT、14C-UBT中不同菌量和時(shí)間的相互關(guān)系。 探究影響幽門螺桿菌小鼠呼氣試驗(yàn)?zāi)Ｐ偷纳镆蛩兀_定影響呼氣試驗(yàn)的小鼠體內(nèi)菌種的組織定位，分離培養(yǎng)并篩選影響呼氣試驗(yàn)的小鼠體內(nèi)尿素酶陽(yáng)性細(xì)菌；對(duì)分離出的尿素酶陽(yáng)性細(xì)菌進(jìn)行飛行質(zhì)譜鑒定及16srDNA-PCR擴(kuò)增測(cè)序，兩種方法確定菌株菌種；對(duì)分離出的尿素酶陽(yáng)性細(xì)菌進(jìn)行尿素酶活性進(jìn)行定性定量的檢測(cè)。 結(jié)果： 小鼠13C-UBT、14C-UBT同一呼氣時(shí)間檢測(cè)數(shù)值與小鼠飼喂藥量成正相關(guān)；同一飼喂藥量中隨著飼喂藥物后檢測(cè)時(shí)間延長(zhǎng)，衡量呼氣的檢測(cè)數(shù)值增大，一定時(shí)間后達(dá)到最大值，此后隨時(shí)間的延長(zhǎng)，檢測(cè)數(shù)值減小。 成功建立幽門螺桿菌小鼠13C-UBT、14C-UBT模型：13C-UBT模型中選用0.1mg-13C-Urea藥量，給予小鼠300ml活動(dòng)空間，飼喂藥物后呼氣10min，收集250ml氣體進(jìn)行檢測(cè)；14C-UBT呼氣中選用0.556KBq-14C-Urea藥量，給予200ml活動(dòng)空間，飼喂藥物后呼氣10min，收集150ml氣體進(jìn)行檢測(cè)；飼喂小鼠不同菌量H.pylori進(jìn)行相同呼氣時(shí)間的檢測(cè)，13C-Urea、14C-Urea呼氣試驗(yàn)中呼氣數(shù)值均隨著H.pylori菌量的升高而增大；飼喂小鼠不同H.pylori菌量，13C-UBT、14C-UBT各曲線走勢(shì)相同，低濃度的菌量受到小鼠體內(nèi)因素的影響大，1×109CFU的H.pylori菌量有很好的增幅曲線和趨勢(shì)。 成功對(duì)小鼠呼氣試驗(yàn)中參與細(xì)菌進(jìn)行定位，參與影響呼氣試驗(yàn)的細(xì)菌位于胃及小腸組織，篩選出155株尿素酶陽(yáng)性細(xì)菌；利用飛行質(zhì)譜及16srDNA-PCR擴(kuò)增測(cè)序兩種方法確定菌株菌種，大部分為大腸桿菌，剩余以侵肺巴斯德菌居多，也檢測(cè)出緩慢葡萄球菌、流感嗜血桿菌、鏈球菌等細(xì)菌；通過對(duì)分離出的細(xì)菌進(jìn)行尿素酶活性定性定量測(cè)定，分離出的細(xì)菌尿素酶活性遠(yuǎn)遠(yuǎn)小于H.pylori。 結(jié)論： （1）成功建立H.pylori小鼠13C-UBT、14C-UBT模型，能夠?qū)π∈筮M(jìn)行無創(chuàng)性實(shí)時(shí)檢測(cè)； （2）小鼠13C-UBT、14C-UBT模型中13C-Urea、14C-Urea飼喂藥量、 H.pylori菌量、呼氣檢測(cè)時(shí)間均與檢測(cè)數(shù)值相關(guān)。其中H.pylori低濃度菌量受小鼠體內(nèi)因素的影響大，H.pylori菌量為1×109CFU受到的影響��； （3）雖分離出的影響呼氣試驗(yàn)的細(xì)菌尿素酶活性小于H.pylori，但在進(jìn)行小鼠呼氣試驗(yàn)中仍要注意并盡量減少其對(duì)實(shí)驗(yàn)的影響。
[Abstract]:Objective: To study the conditions and methods of constructing the mouse exhale model by establishing the model of 13C-UBT and 14C-UBT in Helicobacter pylori, and to provide a non-invasive real-time detection method for the study of Helicobacter pylori animal model. Objective To evaluate the immune protective effect of the HP vaccine in the mouse model and to provide a simple, rapid, cheap and reliable diagnostic method for the in-depth study of the pathogenicity of the H. pylori. Methods: The method of constructing the exhale model of mice by using BABL/ c mice at 6 to 8 weeks was used to establish the experimental model of 13C-UBT and 14C-UBT by feeding different doses of 13C-Urea and 14C-Urea in mice. In the same time, the exhale relation of the mice fed with different bacteria was investigated in the same detection time. The mice were fed with different amounts of H. pylori and different time, and the amount and time of different bacteria in the 13C-UBT, 14C-UBT were explored. To explore the biological factors that affect the exhale test model of Helicobacter pylori, determine the tissue location of the strain in the mice that affect the breath test, isolate the culture, and screen the mice that affect the exhale test. and carrying out flight mass spectrum identification and 16srDNA-PCR amplification sequencing on the isolated urea-enzyme-positive bacteria, and determining the strain strains by two methods; and carrying out urease activity on the isolated urea-enzyme-positive bacteria, line characterization The results showed that the same exhale time of 13C-UBT and 14C-UBT in mice was positively correlated with that of the mice. the maximum value is reached, The detection value was decreased with the extension of time. The model of 13C-UBT and 14C-UBT was successfully established in the model of 13C-UBT and 14C-UBT in the model of 13C-UBT, and 300ml of the active space was given. After the drug was fed, it was exhale for 10min, and 250ml of the gas was collected for testing; and 0.556KBq-14C-Urea was used in the 14C-UBT exhale, and 200ml was given. The activity space, after the drug was fed for 10 minutes, collected 150ml of gas for testing; the same expiration time was detected by H. pylori in the feeding mice, and the exhale value in the 13C-Urea and 14C-Urea expired test increased with the increase of the amount of H. pylori, and the amount of H. pylori in the feeding mice was 13C-U. The curves of BT and 14C-UBT were the same, and the amount of the low-concentration bacteria was affected by the internal factors of the mice. There was a good growth curve and trend in the amount of Lori. The bacteria were located in the breath test of the mice successfully. The bacteria involved in the breath test were located in the stomach and small intestine, and 155 strains of urease-positive bacteria were screened out. Using the flight mass spectrometry and the 16srDNA-PCR amplification test, the bacteria were screened out. The strain is determined by two methods, most of which are E. coli, and the rest of the strain is the Pasteurella, and the bacteria such as the slow Staphylococci, the Haemophilus influenzae and the streptococcus are also detected; and the isolated bacteria are subjected to the qualitative and quantitative determination of the activity of the urease, and the strains are separated out. bacteria The activity of urease is much less than that of H. pylori. Conclusion: (1) H. pylori mouse 13C-UB was successfully established. T, 14C-UBT model, capable of non-invasive real-time detection of mice; (2) 13C-Urea, 14C-Urea feeding in mouse 13C-UBT, 14C-UBT model The amount of H. pylori and the detection time of H. pylori were related to the detection value. The amount of H. pylori in the mice was affected by the in vivo factors. (3) The activity of bacterial urease was less than that of H. py.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R-332
【參考文獻(xiàn)】
相關(guān)期刊論文 前10條
1 樊新霞 ,姬珊;幽門螺桿菌致病機(jī)制的研究進(jìn)展[J];臨床醫(yī)學(xué);2003年04期
2 任英霞;崔東來;張玉琢;;熒光定量聚合酶鏈反應(yīng)在胃黏膜幽門螺桿菌感染診斷中的應(yīng)用研究[J];臨床薈萃;2008年11期
3 李巖;幽門螺桿菌檢測(cè)方法[J];中國(guó)實(shí)用內(nèi)科雜志;2002年01期
4 胡伏蓮;胡品津;劉文忠;王繼德;呂農(nóng)華;蕭樹東;張萬岱;成虹;謝勇;;第三次全國(guó)幽門螺桿菌感染若干問題共識(shí)報(bào)告[J];胃腸病學(xué);2008年01期
5 閆偉;曹建彪;;胃幽門螺桿菌檢測(cè)技術(shù)進(jìn)展[J];世界華人消化雜志;2009年15期
6 張萬岱;幽門螺桿菌若干問題的共識(shí)意見[J];中華消化雜志;2000年02期
7 李曉波,劉文忠;幽門螺桿菌常用診斷方法的評(píng)價(jià)[J];中華消化雜志;2002年11期
8 張萬岱,徐智民;幽門螺桿菌感染診斷方法的評(píng)價(jià)與診斷標(biāo)準(zhǔn)[J];中華全科醫(yī)師雜志;2004年06期
9 辛強(qiáng);;改良Warthin-Starry銀染色法在幽門螺桿菌檢測(cè)中的應(yīng)用[J];中國(guó)衛(wèi)生檢驗(yàn)雜志;2008年11期
10 ;Stability of randomly amplified polymorphic DNA fingerprinting in genotyping clinical isolates of Helicobacter pylori[J];World Journal of Gastroenterology;2003年09期
本文編號(hào)：2351851
本文鏈接：http://sikaile.net/xiyixuelunwen/2351851.html
[Abstract]:Objective: To study the conditions and methods of constructing the mouse exhale model by establishing the model of 13C-UBT and 14C-UBT in Helicobacter pylori, and to provide a non-invasive real-time detection method for the study of Helicobacter pylori animal model. Objective To evaluate the immune protective effect of the HP vaccine in the mouse model and to provide a simple, rapid, cheap and reliable diagnostic method for the in-depth study of the pathogenicity of the H. pylori. Methods: The method of constructing the exhale model of mice by using BABL/ c mice at 6 to 8 weeks was used to establish the experimental model of 13C-UBT and 14C-UBT by feeding different doses of 13C-Urea and 14C-Urea in mice. In the same time, the exhale relation of the mice fed with different bacteria was investigated in the same detection time. The mice were fed with different amounts of H. pylori and different time, and the amount and time of different bacteria in the 13C-UBT, 14C-UBT were explored. To explore the biological factors that affect the exhale test model of Helicobacter pylori, determine the tissue location of the strain in the mice that affect the breath test, isolate the culture, and screen the mice that affect the exhale test. and carrying out flight mass spectrum identification and 16srDNA-PCR amplification sequencing on the isolated urea-enzyme-positive bacteria, and determining the strain strains by two methods; and carrying out urease activity on the isolated urea-enzyme-positive bacteria, line characterization The results showed that the same exhale time of 13C-UBT and 14C-UBT in mice was positively correlated with that of the mice. the maximum value is reached, The detection value was decreased with the extension of time. The model of 13C-UBT and 14C-UBT was successfully established in the model of 13C-UBT and 14C-UBT in the model of 13C-UBT, and 300ml of the active space was given. After the drug was fed, it was exhale for 10min, and 250ml of the gas was collected for testing; and 0.556KBq-14C-Urea was used in the 14C-UBT exhale, and 200ml was given. The activity space, after the drug was fed for 10 minutes, collected 150ml of gas for testing; the same expiration time was detected by H. pylori in the feeding mice, and the exhale value in the 13C-Urea and 14C-Urea expired test increased with the increase of the amount of H. pylori, and the amount of H. pylori in the feeding mice was 13C-U. The curves of BT and 14C-UBT were the same, and the amount of the low-concentration bacteria was affected by the internal factors of the mice. There was a good growth curve and trend in the amount of Lori. The bacteria were located in the breath test of the mice successfully. The bacteria involved in the breath test were located in the stomach and small intestine, and 155 strains of urease-positive bacteria were screened out. Using the flight mass spectrometry and the 16srDNA-PCR amplification test, the bacteria were screened out. The strain is determined by two methods, most of which are E. coli, and the rest of the strain is the Pasteurella, and the bacteria such as the slow Staphylococci, the Haemophilus influenzae and the streptococcus are also detected; and the isolated bacteria are subjected to the qualitative and quantitative determination of the activity of the urease, and the strains are separated out. bacteria The activity of urease is much less than that of H. pylori. Conclusion: (1) H. pylori mouse 13C-UB was successfully established. T, 14C-UBT model, capable of non-invasive real-time detection of mice; (2) 13C-Urea, 14C-Urea feeding in mouse 13C-UBT, 14C-UBT model The amount of H. pylori and the detection time of H. pylori were related to the detection value. The amount of H. pylori in the mice was affected by the in vivo factors. (3) The activity of bacterial urease was less than that of H. py.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R-332
【參考文獻(xiàn)】
相關(guān)期刊論文 前10條
1 樊新霞 ,姬珊;幽門螺桿菌致病機(jī)制的研究進(jìn)展[J];臨床醫(yī)學(xué);2003年04期
2 任英霞;崔東來;張玉琢;;熒光定量聚合酶鏈反應(yīng)在胃黏膜幽門螺桿菌感染診斷中的應(yīng)用研究[J];臨床薈萃;2008年11期
3 李巖;幽門螺桿菌檢測(cè)方法[J];中國(guó)實(shí)用內(nèi)科雜志;2002年01期
4 胡伏蓮;胡品津;劉文忠;王繼德;呂農(nóng)華;蕭樹東;張萬岱;成虹;謝勇;;第三次全國(guó)幽門螺桿菌感染若干問題共識(shí)報(bào)告[J];胃腸病學(xué);2008年01期
5 閆偉;曹建彪;;胃幽門螺桿菌檢測(cè)技術(shù)進(jìn)展[J];世界華人消化雜志;2009年15期
6 張萬岱;幽門螺桿菌若干問題的共識(shí)意見[J];中華消化雜志;2000年02期
7 李曉波,劉文忠;幽門螺桿菌常用診斷方法的評(píng)價(jià)[J];中華消化雜志;2002年11期
8 張萬岱,徐智民;幽門螺桿菌感染診斷方法的評(píng)價(jià)與診斷標(biāo)準(zhǔn)[J];中華全科醫(yī)師雜志;2004年06期
9 辛強(qiáng);;改良Warthin-Starry銀染色法在幽門螺桿菌檢測(cè)中的應(yīng)用[J];中國(guó)衛(wèi)生檢驗(yàn)雜志;2008年11期
10 ;Stability of randomly amplified polymorphic DNA fingerprinting in genotyping clinical isolates of Helicobacter pylori[J];World Journal of Gastroenterology;2003年09期
本文編號(hào)：2351851
本文鏈接：http://sikaile.net/xiyixuelunwen/2351851.html
最近更新
教材專著
