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  • 學位論文

KIF9重組蛋白片段不適合作為婦科疾病的自體抗體診斷標記

KIF9 Recombinant protein fragment is not suitable as a diagnostic autoantibody marker of gynecological diseases

指導教授 : 陳凌雲

摘要


婦科疾病:子宮肌腺症(Adenomyosis)、子宮肌瘤(Myoma)和子宮內膜異位症(Endometriosis),雖然不是造成高死亡率的疾病,但若症狀過於嚴重,不僅會影響患者的生活品質甚至影響到患者的生育能力,患者接受治療後其復發率仍偏高,目前臨床上唯一可用來確定診斷這三種疾病的檢查只有侵入性較高的腹腔鏡檢查。文獻指出SEREX (Serological identification of antigens by recombinant expression cloning)的實驗方法,利用癌症患者血清中含有因癌變過程所產生的自體抗體來協助診斷或當成癒後治療追蹤的工具。在偵測子宮肌腺症、子宮肌瘤和子宮內膜異位症的診斷標記中,如果能藉由抽血檢查血清中有效的抗腫瘤抗原自體抗體,將會對患者在診斷及治療追蹤上有所助益。先前本實驗室以酵素連結免疫吸附分析法已經找到,以IMP1與cyclin B1自體抗體同時檢測婦女是否罹患子宮內膜異位瘤,其靈敏度有83.9% 特異性為72.7%;因此我們希望能再找看看是否有其他針對子宮肌腺症、子宮肌瘤和子宮內膜異位症的特異性TAA存在,可當成診斷的標記。以酵素連結免疫吸附分析法與西方墨點法篩檢KIF9腫瘤相關抗原的自體抗體,發現pET32a載體所表達的thioredoxin蛋白會干擾偵測結果而呈現偽陽性;改以pET30a載體表達KIF9腫瘤相關抗原重組蛋白片段,結果在84位婦科疾病患者血清中,陽性率只有1.2%,顯示KIF9重組蛋白片段不適合作為婦科疾病的診斷標記。

並列摘要


Gynecological diseases such as adenomyosis, myoma, and endometriosis, although are not diseases with high mortality rates, if the symptoms are too severe, they will not only affect the patients` quality of life, but also result in infertility of the patients. Even after the patients have received treatment, the recurrence rate is still high. Currently, the only available clinical diagnostic examination of these three diseases is the highly invasive procedure-laparoscopy. References reflect the SEREX (Serological identification of antigens by recombinant expression cloning) experimental method, which identifies TAAs by screening the patient`s sera, produces auto-antibodies by the tumorgenesis process. It would be helpful in diagnosis and treatment if we can detect the autoantibodies in the patients` serum as diagnostic or prognosis markers for gynecological diseases. Previously, in our laboratory, it has been found that the use of IMP1 and cyclin B1 autoantibody coordination can aid the detection of endometrioma. The sensitivity and specificity was 83.9% and 72.7%. Because of the above results, we hope to find other specific TAAs to use as autoantiboby diagnostic markers for adenomyosis, myoma and endometriosis. By using ELISA and the Western blot experiment to screen KIF9 autoantiboby, we have found that pET32a vector recombinant protein expressions would produce false positive results by inducing thioredoxin protein influence. Through the usage of pET30a vector instead of pET32a to express KIF9 tumor associated protein, out of 84 patients with gynecological diseases, only 1.2% of tested serum showed positive reactions. This indicates KIF9 recombinant protein fragment is not suitable as a diagnostic autoantibody marker of gynecological diseases.

參考文獻


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