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標題: 利用突變特異性擴增系統偵測貓多囊性腎病與應用RIFLE分級評估犬隻急性腎衰竭預後
Molecular detection of feline polycystic kidney disease by multiplex amplification refractory mutation system-PCR and Prognosis of acute kidney injury in dogs using RIFLE criteria
作者: 李雅珍
Lee, Ya-Jane
關鍵字: canine;犬;feline;cystic kidney;renal failure;貓;囊腎;腎衰竭
出版社: 獸醫學系暨研究所
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在第二章我們建立了新的突變特異性擴增系統(PCR-ARMS)來偵測貓多囊性腎病的PKD1 基因點突變,在嘗試了多種引子(primer)的組合後,以三引子在同一管內反應進行基因增幅,不但可成功的偵測此基因的點突變,且可避免偽陰性的發生。分別對35隻已以基因定序確定基因序列的貓(15隻突變型,20隻非突變型)進行測試,發現無論敏感性和專一性都可達到百分之百。結果顯示此方法不但具有便利和花費較低的優勢,也可準確的偵測貓多囊性腎病的PKD1 基因點突變。
第三章中,我們則嘗試將人醫最近發展出的急性腎損傷(acute kidney injury)分類方法RIFLE (risk, injury, failure, loss and end-stage) criteria,應用在犬隻的急性腎損傷病例。利用853隻患有急性腎損傷的犬隻進行回溯性測試,將病例以RIFLE criteria 由輕微至嚴重分級後,以Risk 組為比較基準組,結果顯示無論是Injury 或Failure組都有顯著較高的死亡率(p < 0.05)。另外,我們還利用了多重性回歸分析在犬隻急性腎損傷的病例中找出和死亡相關的兩個因子,血磷和下痢情形。以此二因子加上RIFLE criteria產生一個新的算式。經由算式計算每一個病例的分數,以receiver operating characteristic (ROC) curve分析,以ROC面積大小來評估專一性和敏感性,結果顯示新算式比起單純的RIFLE criteria可更準確的評估預後。

In this study, autosomal dominant polycystic kidney disease (ADPKD) in feline and the prognosis of acute renal failure in canine were the main topics to be investigated. In chapter I, results of recent studies in both human and feline ADPKD as well as the methods for evaluating the prognosis of renal failure were introduced.
In chapter two, establishment of a molecular method derived from multiplex amplification refractory mutation system-PCR (ARMS-PCR) for detection of feline ADPKD was described. By using three primers, ARMS-PCR successfully detected the point mutation of exon 29 in feline PKD1 gene from 35 feline blood samples, including 15 mutant cats and 20 wild type cats. The result was validated with that of automated sequencing; 100% of specificity was revealed, indicating that this method is not only advantaged with convenience and low cost but also with high sensitivity and specificity.
In chapter three, we tried to use the RIFLE (risk, injury, failure, loss and end-stage) criteria, derived from human medicine, to evaluate prognosis of 853 dogs with acute kidney injury (AKI) by retrospective study. Taking the dogs in class Risk as the reference, mortality rate of the dogs either in class Injury or in class Failure was significantly higher than that of the dogs in class Risk (p < 0.05). In addition, we also generated a new score, which was combined with RIFLE criteria with other two factors, i.e. the level of serum phosphorus and diarrhea. Consequently, the statistic results revealed that the RIFLE criteria could be applied in veterinary medicine for classification of dogs with AKI and the new score had a better performance in prediction of prognosis than RIFLE criteria alone.
In chapter four, conclusions and perspectives of our study were described. The application of the new diagnostic method or the evaluation criteria derived from human medicine can benefit veterinarians and also provide animal model for human medicine.
其他識別: U0005-3006201014453900
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