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The Investigation of Women's Recognition Tendency of Pap Smear Services and the Influent Factors of Its Acceptability
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A Study on Training Evaluation for Cultivate-Education of State-Run-Corporation-Taking Taiwan Power Company for an Example
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Assessment of the efficacy of platelet transfusion with post-transfusion corrected count increment(CCI)
No full text[[abstract]]許多疾病都可能會造成血小板減少，而輸注血小板是面對這種併發症的主要處置方法。止血是血小板的重要生理功能，為了維持此功能必須要有足夠質量的血小板與凝血因子共同作用方可達成。血小板輸血後是否能有效地增加血小板數量是影響臨床治療成效的重要指標。利用校正後血小板增加數量(corrected count increment；CCI值)作為血小板的輸血療效評估是目前能得到較客觀及正確的重要參考指標。
在 2009年從4月至10月間, 我們收集47位輸注血小板的病患參與本研究，藉由探討病患輸注血小板的CCI值與各項因素間之相關性，其中包括性別、年齡、血小板成分、白血球過濾器、血型相容性、輸血反應以及不規則抗體，以了解台灣人輸注血小板之療效。我們收集輸注血小板後18至24小時的血小板數換算成CCI值,其結果以大於或等於 4,500 m2/μL判定成功的有效輸血.我們的研究結果顯示平均CCI值是6,777m2/μL (-52,903 m2/μL~119,653m2/μL)。在研究對象中有效輸血僅占了51.06%。然而，國人平均體表面積與外國人平均體表面積有明顯差異。因此，本研究結果建議應定義出適合國人之CCI參考值,以提升血小板輸注成效並節省醫療成本。
Many diseases may lead to thrombocytopenia, and platelet transfusion for this complication is the main treatment. Hemostasis is an important physiological function of platelet. To maintain this function, there must be enough quantity and quality of platelet to interact with coagulation factors. Whether platelet transfusion can effectively increase the quantity of platelet is an important indicator of the efficacy. Corrected count increment (CCI value) as the evaluation of efficacy of platelet transfusion, can be an objective and correct reference.
From April to October in 2009, we included 47 patients with platelet transfusion in this study. We investigated the correlation between CCI value of post-transfusion and several factors, including gender, age, platelets component, WBC filtration, blood type compatibility, transfusion reaction, and irregular antibodies, to understand the efficacy of platelet transfusion in Taiwan. We analyzed platelets count after transfusion of 18 to 24 hours to calculate the CCI value. The outcome was whether or not a transfusion lead to a successful response, defined as a CCI greater or equal to 4,500 m2/μL.
Our results showed that the mean value of CCI value is 6,777 m2/μL (-52,903 m2/μL~119,653 m2/μL). Effectiveness of platelet
transfusion is only 51.06%. However, the average body surface area (BSA) of Taiwanese is different from other populations. Therefore, the results of this study conclude that the necessity of further study to define a reference value of CCI for Taiwanese to enhance the effectiveness of platelet transfusion and to save medical costs
The development of detecting techniques of Xanthomonas axonopodis pv. dieffenbachiae and X. oryzae pv. oryzae
Get PDF[[abstract]]Xanthomonas屬為革蘭氏陰細菌且是世界重要植物病原菌之一，可造成許多農業損失。其中火鶴花細菌性葉枯病菌及水稻白葉枯病菌會感染許多台灣地區之經濟作物，為能早期鑑定出火鶴花細菌性葉枯病菌及水稻白葉枯病菌，本研究利用gyrB、virD4、rpoD及Lytic eznyme等序列並結合PCR開發快速之檢測技術。研究中將火鶴花細菌性葉枯病菌、水稻白葉枯病菌、檬果細菌性黑斑病菌、茄科細菌性斑點病菌、十字花科黑腐病菌及柑桔潰瘍病菌基因體中之gyrB、virD4、rpoD及Lytic eznyme等序列進行核酸選殖，利用生物資訊進行多序列比對顯示序列間之差異性，並進一步利用火鶴花細菌性葉枯病菌及水稻白葉枯病菌之序列差異處設計專一性引子組搭配聚合酶連鎖反應技術進行引子組之鑑定。在偵測火鶴花細菌性葉枯病菌方面，分別使用XAD-virD4、XAD-gyrB及XAD-LE等3組專一性引子組，其PCR檢測之專一性皆為100％，其中XAD-gyrB引子組可偵測之敏感度介於40pg與4pg之DNA濃度或103cfu與102cfu之間，而XAD-virD4引子組可偵測之敏感度介於2pg與200fg之DNA濃度或102cfu與101cfu之間，至於XAD-LE引子組其偵測之敏感度介於30pg與3pg 之DNA濃度或102cfu與101cfu之間。在偵測水稻白葉枯病菌方面，分別使用XOO-gyrB及XOO-rpoD等2組專一性引子組，其PCR檢測之專一性皆為100％；其中XOO-gyrB引子組可偵測之敏感度介於30pg與3pgDNA濃度或102cfu與101cfu之間，而XOO-rpoD引子組可偵測之敏感度介於40pg與4pg之DNA濃度或103cfu與102cfu之間。此外，研究中利用多引子組PCR(Multiplex PCR)之技術來偵測火鶴花細菌性葉枯病或水稻白葉枯病菌。以引子組XAD-virD4、XAD-gyrB及XAD-LE之組合用於偵測火鶴花細菌性葉枯病；引子組XOO-gyrB及XOO-rpoD之組合用於偵測水稻白葉枯病菌，其結果顯示引子組之組合專一性皆為100％。因此可知，PCR及多引子組PCR為可使用及可靠之偵測技術，可用於篩選及偵測火鶴花細菌性葉枯病菌及水稻白葉枯病菌之診斷工具。
Xanthomonas is a Gram-negative bacterium and one of the most important phytopathogenic bacteria in the world, which causes many agricultural loss. As Xanthomonas axonopodis pv. dieffenbachiae (Xad) and Xanthomonas oryzae pv. oryzae (Xoo) affected many economic plants in Taiwan. In order to identify Xad and Xoo at the early stage of infection, we plan to develop a rapid detecting technique based on the DNA sequences of gyrB、virD4、rpoD and Lytic enzyme. We cloned several DNA fragments from different species of Xad、Xoo、Xcm、Xav、Xcc and Xac, and the bioinformatics analysis of Multiple Sequences Alignment displayed the diversity sequences. The specific primer-pairs were designed for the PCR identification of Xad and Xoo by the diversity sequences. On the detection of Xad, the PCR detection results of XAD-gyrB、XAD-virD4 and XAD-LE revealed the specificity will be 100％. The sensitivity of XAD-gyrB the will be 40 pg-4 pg DNA or 103 cfu-102 cfu. The sensitivity of XAD-virD4 will be 2 pg-200 fg DNA or 102-101 cfu. The sensitivity of XAD-LE will be 30 pg-3 pg DNA or 102 cfu-101 cfu. On the detection of Xoo, the PCR detection results of XOO-gyrB、XOO-rpoD the specificity will be 100％. The sensitivity of XOO-gyrB the sensitivity will be 30 pg-3 pg DNA or 102 cfu-101 cfu. The sensitivity of XOO-rpoD the sensitivity will be 40 pg-4 pg DNA or 103 cfu-102 cfu. Moreover, we used Multiplex-PCR method to detect Xad or xoo. The combination of XAD-virD4、XAD-gyrB and XAD-LE was used to detect Xad and the combination XOO-gyrB and XOO-virD4 was used to detect Xoo. The results revealed the specificity of these combinations will be 100% for detection. Accordingly, the PCR and multiplex PCR are useful and reliable methods for the detection. This technique could be a diagnostic tool for screening and monitoring of Xanthomonas axonopodis pv. dieffenbachiae (Xad) and Xanthomonas oryzae pv. oryzae (Xoo)
PCR techniques for identification of phytopathogenic bacteria in Xanthomonas
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中文摘要…………………………………………………………… I
英文摘要…………………………………………………………… Ⅱ
壹、前人研究……………………………………………………… 1
貳、材料與方法…………………………………………………… 1
一、 供試菌株之來源、培養與保存……………………………… 11
二、 抗體之製備………………………..………………………… 11
三、 供試菌株染色體DNA之純化……………………………… 11
四、 共通性引子組之設計……………………………………… 11
五、 聚合酵素連鎖反應(polymerase chain reaction PCR)… 12
六、 PCR產物之純化…………………………………………… 12
七、 PCR產物與質體之接合作用……………………………… 12
八、 核酸轉型作用……………………………………………… 13
九、 DNA定序與引子組設計…………………………………… 13
十、 單一引子組之專一性子檢測……………………………… 14
十一、 單一引子組之敏感度分析……………………………… 14
十二、 多引子組之專一性子檢測……………………………… 15
十三、 IgG抗體的純化………………………………………… 16
十四、 磁珠結合抗體之操作………………………………… 16
十五、 免疫磁珠之病原細菌分離法…………………………… 17
參、結果…………………………………………………………… 18
一、 XAC、XCC及XAV菌株之ITS核酸定序與分析………… 18
(一) XAC菌株之ITS核酸定序與分析……………………… 18
(二) XCC菌株之ITS核酸定序與分析……………………… 18
(三) XAV菌株之ITS核酸定序與分析……………………… 19
(四) XAC、XCC及XAV之ITS核酸定序與分析…………… 19
二、 XAC、XCC及XAV菌株Lytic enzyme核酸定序與分析 20
三、 單一引子組之專一性分析……………………………… 21
(一) ITS單一引子組之專一性檢測…………………………… 21
1. XCC之專一性檢測……………………………………… 21
2. XAC之專一性檢測……...……………………………… 21
3. XAV之專一性檢測……………………………………… 22
(二) LE單一引子組之專一性檢測…………………… 22
1. XCC之專一性檢測………………………………………... 22
2. XAC之專一性檢測………………...……………………… 22
3. XAV之專一性檢測………………………………………... 23
四、 專一性引子之偵測敏感度分析………………………… 23
(一) XCC專一性引子之偵測敏感度…………………………… 23
1. 引子組XCC-ITS-F/ Xan-16S-R之PCR敏感度分析…...… 23
2. 引子組XCC-LE2-F/ XCC-LE2-R之PCR敏感度分析…… 23
3. 引子組XCC-LE4-F/ Xan-Lytic-R之PCR敏感度分析........ 23
(二) XAC專一性引子之偵測敏感度…………………………… 24
1. 引子組XAC-ITS-F/ XAC-ITS-R之敏感度分析….……… 24
2. 引子組Xan-Lytic-F/ XAC-LE4-R之敏感度分析……….... 24
(三) XAV專一性引子偵測敏感度分析………………………… 24
1. 引子組XAV-ITS-F/ Xan-16S-R之敏感度分析….……….. 24
2. 引子組XAV-LE4-F/ Xan-Lytic-R之敏感度分析.……….. 25
五、 Multiplex PCR在檢測上之應用………………………… 25
(一) 雙引子組對單一DNA之檢測…………………………… 25
(二) 多引子組對單一DNA之檢測…………………………… 25
(三) 單一引子組對多種DNA之檢測………………………… 25
(四) 三引子組對單一DNA之檢測…………………………… 26
(五) 三引子組對三種DNA之檢測…………………………… 26
六、 免疫磁株分離細菌之效果分析…………………………… 27
肆、討論…………………………………………………………… 28
伍、參考文獻……………………………………………………… 33
陸、圖表…………………………………………………………… 39
柒、附錄…………………………………………………………… 69
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Zaccardelli, M., Campanile, F., Spasiano, A., and Merighi, M. 2007. Detection and identification of the crucifer pathogen, Xanthomonas campestris pv. campestris, by PCR amplification of the conserved Hrp/type III secretion system gene hrcC. Eur J Plant Pathol. 118：299-306.[[abstract]]Xanthomonas屬之植物病原細菌為革蘭氏陰性、呈桿狀，具有一條極生鞭毛，通常會引起植物之重要病害。其中柑橘潰瘍病菌、十字花科黑腐病菌及茄科細菌性斑點病菌則為台灣地區常見之植物病原細菌，可引起許多經濟性蔬果之重要病害。為了能在早期鑑定出柑橘潰瘍病原細菌、十字花科黑腐病原細菌及茄科細菌性斑點病原細菌之感染，本研究利用internal transcribe spacer (ITS)與lytic enzyme序列結合PCR開發出快速之檢測技術，研究中將柑橘潰瘍病菌、十字花科黑腐病菌及茄科細菌性斑點病菌基因體中之ITS與lytic enzyme 等DNA進行選殖，利用生物資訊進行多序列分析，顯示不同種之Xanthomonas間的DNA序列具有較多的變異。利用序列之差異性可設計出專一性引子，成功的檢測出柑橘潰瘍病菌、十字花科黑腐病菌及茄科細菌性斑點病菌。在單一引子組PCR(Uniplex PCR)分析中能夠鑑定柑橘潰瘍病菌、十字花科黑腐病菌及茄科細菌性斑點病菌之差異性。研究中也進一步開發出以多引子組PCR (Multiplex PCR)技術來鑑定柑橘潰瘍病菌、十字花科黑腐病菌及茄科細菌性斑點病菌。本研究中也利用免疫磁珠分離(IMS)搭配PCR技術來達到分離及鑑定病原細菌之目的，葉片上之柑橘潰瘍病原細菌在免疫磁珠PCR檢測時之敏感度則介於101與102 cfu之間。
Xanthomonas phytopathogenic bacteria were gram-negative, rod-shaped, polarly-flagellated bacteria which members caused serious plant diseases usually. Xanthomonas axonopodis pv. citri (XAC) , Xanthomonas campestris pv. campestris (XCC) and Xanthomonas axonopodis pv. vesicatoria (XAV) affected many economic plants which included vegetables and fruits in Taiwan. In order to identify XAC、XCC and XAV at the early stage of infection, we planed to develop a rapid detecting technique based on specific sequences of the internal spacer regions (ITS) or lytic enzyme. In this study, we cloned several DNA fragments of ITS and lytic enzyme from XAC、XCC and XAV. The bioinformatics analysis by multiple sequences alignment showed the diversity exist in different species of XAC、XCC and XAV. We designed specific primers based on the sequence (ITS and lytic enzyme) divergence and developed the detective method successfully. The development of specific uniplex-PCRs could detect XAC、XCC and XAV. We designed new multiplex-PCRs, could detect XAC、XCC and XAV at the same PCR reaction. In this project, we intended to combine the immuno-magnetic beads separation technique with the specific PCR detecting method to isolate the bacteria. This technique could isolate XAC from leafs and the sensitivity of immuno-magnetic PCR was 101 – 102 cfu