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  本文選題：化膿性腦膜炎 + 腦脊液　； 參考：《寧夏醫(yī)科大學》2013年碩士論文

【摘要】：目的研究化膿性腦膜炎患者腦脊液中致病菌的常見細菌種類、細菌豐度及菌群結(jié)構(gòu)，探討化膿性腦膜炎病原學診斷的新途徑和方法。 方法本研究選取臨床診斷為化膿性腦膜炎患者腦脊液樣本61例，提取化膿性腦膜炎腦脊液細菌基因組DNA、再進行PCR鑒定，同時對CSF進行細菌培養(yǎng)，計算兩種方法的陽性率，將二者陽性率采用SPSS17.0統(tǒng)計軟件進行卡方檢驗。利用16S rDNA宏基因組測序技術(shù)對PCR陽性的樣本進行高通量測序，通過生物信息學分析的方法對測序結(jié)果進行整合、分析。 結(jié)果對選取的61例化膿性腦膜炎腦脊液樣本分別進行細菌培養(yǎng)和檢驗，有26例樣本在550bp處擴增出特異的DNA產(chǎn)物，而細菌培養(yǎng)陽性的樣本僅為10例，61例腦脊液樣本中，26例16S rDNA PCR陽性，，陽性率為42.6%；61例腦脊液樣本中細菌培養(yǎng)陽性為10例，陽性率為16.4%；通過16S rDNA宏基因組測序技術(shù)及生物信息學技術(shù)研究發(fā)現(xiàn)：通過觀察每個樣品所含有的OTU數(shù)量，與其他環(huán)境相比，腦脊液含有的物種種類相對偏少；PM腦脊液樣本致病菌的種類大致可以分為以厚壁菌門和變形桿菌門為主，以屬為分類標準，優(yōu)勢菌屬主要為鏈球菌屬，鮑曼不動桿菌，假單胞菌，奈瑟菌屬等；通過beta多樣性分析, PM腦脊液中細菌感染在屬的水平上兩兩樣本之間差異不大。 結(jié)論通過對上述兩種檢測方法陽性率比較，PCR檢查方法診斷化膿性腦膜炎明顯優(yōu)于分離培養(yǎng)法（P＜0.05）；本研究選取QIAGEN DNeasy Blood and Tissue Kit試劑盒DNA提取的方法，獲得的基因組DNA，引物及PCR條件較好，能對化膿性腦膜炎腦脊液樣本中16S rDNA進行良好的擴增，能適用于化膿性腦膜炎腦脊液宏基因組學的后續(xù)數(shù)據(jù)分析及研究。通過16S rDNA宏基因組測序技術(shù)對于化膿性腦膜炎腦脊液致病菌的種類，豐度及菌群結(jié)構(gòu)的分析發(fā)現(xiàn)銀川地區(qū)感染的優(yōu)勢菌群為：鏈球菌，鮑曼不動桿菌及假單胞菌，還有一些未知的細菌也是造成顱內(nèi)感染的重要元兇。
[Abstract]:Objective to study the common bacteria species, bacterial abundance and bacterial community structure in cerebrospinal fluid (CSF) of patients with suppurative meningitis, and to explore a new approach and method for etiological diagnosis of suppurative meningitis. Methods in this study, 61 samples of cerebrospinal fluid (CSF) from patients with suppurative meningitis were selected to extract bacterial genomic DNA from cerebrospinal fluid (CSF) of suppurative meningitis, then PCR was used to identify CSF, and the positive rate of the two methods was calculated. The positive rates were tested by SPSS17.0 software. High-throughput sequencing of PCR positive samples was carried out by using 16s rDNA macro genome sequencing technique. The results were integrated and analyzed by bioinformatics analysis. Results bacterial culture and examination were carried out in 61 samples of cerebrospinal fluid from suppurative meningitis. Specific DNA products were amplified from 26 samples at 550bp. The positive rate of bacterial culture was 42.6%. The positive rate of bacterial culture was 42.6%, and the positive rate of bacterial culture was 10 cases in 61 cerebrospinal fluid samples. The positive rate was 16. 4%. By studying 16s rDNA macro genome sequencing and bioinformatics, we found that by observing the amount of OTU in each sample, compared with other environments, The species contained in cerebrospinal fluid (CSF) were relatively few. The species of pathogenic bacteria in CSF samples of PM could be roughly divided into two groups: phylum platyphylum and phylum Proteus. The dominant species were Streptococcus and Acinetobacter baumannii. Pseudomonas, Neisseria and so on. By beta diversity analysis, bacterial infection in PM cerebrospinal fluid had little difference between two samples at the generic level. Conclusion the positive rate of the above two methods is better than that of the culture method (P < 0.05) in the diagnosis of suppurative meningitis, and the method of DNA extraction with QIAGEN DNeasy Blood and Tissue Kit kit is selected in this study. The obtained genomic DNA, primers and PCR conditions were good for amplification of 16s rDNA in cerebrospinal fluid samples of suppurative meningitis, and suitable for subsequent data analysis and research on macrogenomics of cerebrospinal fluid in suppurative meningitis. By analyzing the species, abundance and microflora structure of cerebrospinal fluid pathogenic bacteria of suppurative meningitis by 16s rDNA macro genome sequencing, it was found that the dominant bacterial groups of infection in Yinchuan area were Streptococcus, Acinetobacter baumannii and Pseudomonas. Unknown bacteria are also responsible for intracranial infections.
【學位授予單位】：寧夏醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R515.2

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