本文選題：曼氏裂頭蚴 + 病原診斷��； 參考：《中南大學(xué)》2012年碩士論文

【摘要】：[目的]系統(tǒng)觀察和分析曼氏裂頭蚴的解剖組織學(xué)和組織病理學(xué)用以補充對該病原診斷基礎(chǔ)的不足；探索檢測切片中曼氏裂頭蚴抗原和基因的方法用以解決病理切片中不能確定蟲種的鑒定技術(shù)問題。 [方法]收集來自青蛙和病人的曼氏裂頭蚴標(biāo)本42個、來自動物的其他蠕蟲標(biāo)本13個和人體組織標(biāo)本2個。用倒置顯微鏡比較觀察裂頭蚴各部位的解剖結(jié)構(gòu)組織學(xué)特征；福爾馬林固定和石蠟包埋的曼氏裂頭蚴切片標(biāo)本,對HE染色的蟲體不同部位組織學(xué)進(jìn)行比較觀察；制備曼氏裂頭蚴免疫血清,用間接酶免疫組化法比較檢測不同蟲種切片組織中抗原；設(shè)計作曼氏裂頭蚴mtDNACOX1基因部分片斷檢測的引物4對,用商品化試劑分別提取數(shù)種蠕蟲基因組DNA作PCR,取擴增產(chǎn)物經(jīng)電泳觀察目的條帶,并測序,用blast軟件分析其與已知基因序列的符合率。 [結(jié)果]曼氏裂頭蚴顯微解剖組織學(xué)特征為：頭部吸槽內(nèi)具囊腔；體壁具凹凸橫皺褶；體內(nèi)實質(zhì)呈網(wǎng)狀纖維結(jié)構(gòu),富含石灰小體,在體中后部可見排泄小管；從蟲體劃破處可見有大量的呈圓形或橢圓形、無細(xì)胞結(jié)構(gòu)的強折光盤狀體溢出,即石灰小體,其個體明顯比體細(xì)胞大。曼氏裂頭蚴組織病理學(xué)特征為：可因切取部位不同而有差異；頭部形態(tài)最不規(guī)則,其橫切面可見囊腔和成石灰小體；體部和尾部斷面較大；體部的不同切面在組織結(jié)構(gòu)上有差異；除頭部外,蟲體各斷面的共同特征為體壁具凹凸不等的橫皺褶,皮層較厚呈嗜伊紅深染,有的皮層外可見長短不一微毛,體內(nèi)為網(wǎng)狀疏松的實質(zhì)組織,無體腔和消化器官結(jié)構(gòu),可見分散的肌纖維和圓形,橢圓形或不規(guī)則形石灰小體,亦可見排泄管腔；在病理切片9例中有5例在蟲體組織內(nèi)未見典型的網(wǎng)狀結(jié)構(gòu)(呈鈣化變性),石灰小體和肌纖維束結(jié)構(gòu)不典型。用酶免疫組化法最佳條件,對已知并殖吸蟲、豬囊尾蚴和曼氏裂頭蚴的組織切片檢測均未出現(xiàn)假陰性和假陽性,對12例臨床來源的病理切片中裂頭蚴檢測均呈陽性反應(yīng)。在基因擴增用的4對引物中,有兩對(F650/R800和F965/R1120)可檢出曼氏裂頭蚴特異性目的條帶。 [結(jié)論]本研究描述的曼氏裂頭蚴顯微解剖組織學(xué)特征與其觀察方法為病理學(xué)對石蠟包埋前蟲體鑒定提供了簡捷手段；系統(tǒng)觀察和闡述的裂頭蚴組織病理學(xué)特征為病理學(xué)觀察提供了參考依據(jù)；探索檢測曼氏裂頭蚴特異性抗原和基因的方法為病理切片中疑似蟲體鑒定提供了技術(shù)。
[Abstract]:[objective] to observe and analyze systematically the anatomical histology and histopathology of caterpillars mansoni in order to supplement the deficiency of the basis of diagnosis of the pathogen. To explore the method of detecting antigen and gene of mitomycaria mansoni in order to solve the problem of identification of insect species in pathological sections. Methods 42 specimens of mitomycaria mansoni from frogs and patients, 13 specimens of other worms from animals and 2 specimens of human tissues were collected. The anatomical structure and histological characteristics of each part of the caterpillar were observed by inverted microscope, and the histological characteristics of the different parts of the insect body stained by HE were compared with those of the specimens fixed with formalin and paraffin embedded in the sections of the filariae. To prepare the immune serum of mitomycaria mansoni, to compare and detect the antigens in the sections of different insect species by indirect enzyme immunohistochemical method, and to design 4 pairs of primers for the partial detection of mtDNACOX1 gene fragment of mitomycaria mansoni. Several worm genomic DNA were extracted by commercial reagents as PCR products. The target bands were observed by electrophoresis and sequenced. The coincidence rate of the PCR products with known gene sequences was analyzed by blast software. [results] the microanatomical and histological features of the caterpillar were as follows: there was a cystic cavity in the suction groove of the head, a convoluted transverse fold in the body wall, a reticular fibrous structure in the body, rich in lime bodies, and a excretory tubule in the posterior part of the body. A large number of strongly refractive disklike bodies with circular or elliptical shape and no cell structure were found to overflow from the area of the insect body, that is, calcareous bodies, whose individual size was obviously larger than that of somatic cells. The histopathological features of mitocercariae mansoni are as follows: the shape of the head is the most irregular, the cystic cavity and calcareous bodies can be seen on the transverse section of the head, the section of body and tail is large. In addition to the head, the common features of the different sections of the insect body are the uneven transverse folds of the body wall, the thicker layer of the body and the deep staining of the iridescent, and some of them have different hairs of different lengths outside of the cortex, and the common features of the different sections of the body are that the body wall has different transverse folds, and the cortical layer is thicker and reddish. The body was reticular-loose parenchyma, without the structure of body cavity and digestive organ, with scattered muscle fiber and circular, elliptic or irregular calcareous bodies, and also with excretory lumen. Typical reticular structures (calcified denaturation, calcareous bodies and muscular fiber bundles) were not found in 5 of 9 cases of pathological sections. Under the best condition of enzyme immunohistochemical method, no false negative and false positive staining was found in the sections of known paragonimiasis, cysticercus and cysticercus mansoni, but positive reaction was found in 12 pathological sections of clinical origin. Of the 4 pairs of primers used for gene amplification, two pairs of F650 / R800 and F965 / R1120) could detect the specific target bands of mitomycaria mansoni. [conclusion] the microanatomical and histological characteristics and observation methods described in this study provide a simple and convenient method for the identification of paraffin embedded prebugs by pathology. The histopathological characteristics of mitomycaria provided a reference basis for pathological observation, and a method for detecting specific antigen and gene of mitomycaria mansoni provided a technique for identification of suspected parasites in pathological sections.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R383.35

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