【摘要】：病理切片中各種蠕蟲的鑒定是臨床病理診斷中常見的難題,其原因是到目前為止針對不同蟲種的組織形態(tài)特征未見有系統(tǒng)的形態(tài)描述或圖譜分析,也未建立一種實用于臨床鑒別蟲種的方法。本研究對(肺吸蟲、豬囊蟲和曼氏裂頭蚴)此三種蠕蟲病理組織切片進行了形態(tài)學(xué)比較分析和基因檢測。研究方法：收集多個不同種蠕蟲的石蠟切片標本(來源于動物或臨床患者)和12個臨床病理切片標本(來源于疑似感染此三種蠕蟲的臨床患者)；在光學(xué)鏡下比較觀察和分析已知肺吸蟲、豬囊蟲和裂頭蚴的組織結(jié)構(gòu)相似性和特征性,闡述了蟲體組織退變而呈現(xiàn)為不典型形態(tài)結(jié)構(gòu)的特征；根據(jù)此三種蠕蟲的種特異性基因序列,設(shè)計相應(yīng)PCR引物檢測此三種蠕蟲做種特異性,并探索了從組織內(nèi)蠕蟲的病理白片、染色片和組化片中提取DNA的方法以及提取DNA所需的蟲體組織量,最后檢測該方法是否實用于臨床切片標本。結(jié)果顯示：①三種蠕蟲和其他吸蟲的網(wǎng)狀組織結(jié)構(gòu)和皮層具有相似性；三種蠕蟲的特征性標志,肺吸蟲具有尖刀樣的皮棘、橢圓形的斷面和組織內(nèi)可見管腔器官及吸盤,豬囊蟲斷面大且具有多個不規(guī)則囊腔,裂頭蚴斷面呈長橢圓形可見大量石灰小體；退化變性后的蟲體使其特征性結(jié)構(gòu)有不同程度地消失。②用肺吸蟲(L12/R12和F3/B3)、裂頭蚴(F650/R800)和豬囊蟲(F3/R7)的引物分別對此三種蠕蟲、其他寄生蟲、宿主組織進行PCR擴增后,在相應(yīng)的這三種蠕蟲的泳道中有明顯的目的條帶,而其他寄生蟲和宿主組織均未擴增出條帶。③用改良蛋白酶K消化法提出DNA所需最小組織量：肺吸蟲的為1.50mm2×10μm；曼氏裂頭蚴的為4.87mm2×5μm；豬囊蟲的為5.80mm×5μm.研究結(jié)論：描述了這三種蠕蟲的組織病理學(xué),為臨床病理診斷提供了更可靠的形態(tài)學(xué)依據(jù)；建立了一種快速簡便、特異性高和敏感性高的適用于臨床鑒別診斷此三種蠕蟲的分子生物學(xué)方法。圖23幅,表1個,參考文獻84篇。
[Abstract]:The identification of helminth in pathological sections is a common problem in clinical pathological diagnosis. The reason is that so far, no systematic morphological description or map analysis has been done for the morphological characteristics of different species of insects, nor has a method for clinical identification of insects has been established. * (paragonimiasis, cysticercus cellulosae and Schistosoma mansoni) were studied in this study. Methods: Paraffin section specimens (from animal or clinical patients) and 12 clinical pathological section specimens (from clinical patients suspected to be infected with the three worms) were collected and compared under optical microscope. * the structure and similarity of paragonimiasis, cysticercus cellulosae and larval digits were described, and the characteristics of atypical morphological structure were described. The corresponding PCR primers were designed according to the specific gene sequences of these three species of worms, and the three species of worms were detected. The results showed that: (1) the reticular structure and cortex of the three worms and other trematodes were similar; the characteristic markers of the three worms, paragonimiasis, had sharp cuticular spines and ellipses. There are large number of irregular cavities in the cross section of the cysticercus cellulosae * with a large number of irregular cysts, and a large number of calcareous bodies are seen in the section of the cleft spider. The degenerated degenerated worm has disappearance of its characteristic structure. * (L12/ R12 and F3/B3), F650/R800 and cysticercus cellulosae (F3/R7) (3) After PCR amplification of the primers on the swimming lanes of the three worms, other parasites and host tissues, there were obvious target bands in the swimming lanes of the three worms, but no bands were amplified in other parasites and host tissues. The results are as follows: 4.87mm2 * 5 m; cysticercus cellulosae 5.80mm * 5 m. M. conclusion: the histopathology of these three worms is described, which provides a more reliable morphological basis for clinical pathological diagnosis. A rapid, simple, highly specific and sensitive molecular biology suitable for clinical differential diagnosis of these three worms has been established. There are 23 tables, 1 tables and 84 references.
【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】：2013
【分類號】：R361.2

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