| 摘要: | 正確檢測及鑑定造成感染性疾病的微生物，在環境監測、臨床醫學、生物防禦等議題上變得日益重要，病患若受到細菌感染可能會變成嚴重的感染性疾病，所以快速的鑑定致病原，就可以給予適當治療及使群突發獲得控制。標準化培養方式可鑑別致病原，而藥敏試驗可獲得抗生素藥敏試驗結果。然而微生物培養及藥敏試驗需耗費人力、時間、金錢，更重要的是鑑定致病原微生物的生物化學及血清學試驗，只能鑑別至菌屬的種或血清型等層級，而不能直接說明其致病因素特性。
本論文研究以郭綜合醫院98年4月至10月間內科、外科、婦產科、兒科、泌尿科，胸腔內科，耳鼻喉科、放射腫瘤科之住院或門診病人，共917人為採樣對象，分離出1016株菌株，共鑑定出53種菌種，其中革蘭氏陰性菌38種，共726株，革蘭氏陽性菌15種，共290株。由於 rDNA基因是理想的遺傳指標，且16S-23S rDNA間內轉錄區(intergenic spacer region, ITS) 具高度保留性，但於不同菌種間則具明顯變異性，因此可應用於細菌鑑定，利用內轉錄區核酸序列，做為鑑別臨床菌株的工具，聚合酶連鎖反應 (polymerase chain reaction, PCR) 技術對於挑剔性及無法培養的微生物，提供了獨特的優勢。
本研究利用PCR 技術，可縮短鑑定菌種時效，由24小時縮短為4小時；配合本實驗室每半年的各菌種抗生素感受性試驗統計結果，可將完整報告由需要48小時縮短為只需24小時，可提供臨床醫師初步訊息，得以盡快治療病友。另微生物培養檢查而產生的醫療廢棄物，若改以本研究實驗方法，可使原本使用的傳統生化試驗鑑定法而產生之醫療廢棄物，由每日約6公斤減少至約3.5公斤，大幅減少42%的感染性醫療廢棄物。綜合以上結果，核酸片段分析是快速、簡單且準確度高的方法，可提供臨床菌種鑑定更好的選擇，以促進醫療品質。
The ability to accurately detect and identify microorganisms that are capable of causing infectious diseases has become increasingly important in environmental surveillance, clinical medicine, and biodefense settings. Bacteria infection may emerge as a serious infection for patients. Rapid identification of pathgens is required so that appropriate therapy can be given and outbreaks controlled. Standard culture and susceptibility tests permit pathogen identification and antimicrobial susceptibility profiling, but they are laborious, time-consuming, and expensive and required labile natural products. More importantly, the biochemical and serologic tests that are routingly utilized for pathogen identification only in the species or serogroup level and do not directly characterize virulence factors.
In this study, 1016 bacterial clones were isolated from 917 patients. By biochemical detection, there were 38 gram negative bacteria species and 15 gram positive species. The rDNA genes are ideal genetic targets that can be used for bacterial identification, since these sequences are highly conserved between species. The intergenic spacer (ITS) region between the 16S and 23S rDNA genes is a good candidate for bacterial species identification. ITS sequence-based identification is reliable and provides a promising tool for elucidation of the clinical significance of the different species. Polymerase chain reaction (PCR)-based methods offer a distinct advantage for the detection of fastidious and noncultivable organisms.
By using PCR technology, it can reduce decidable the mold identifiable spawn effectiveness (by 24 hour reduction as 4 hours), coordinate this laboratory every half year display antibiotic sensation experiment results, this method takes advantage of the results and the antibiotic feeling test results, provides clinician the preliminary news, can treat the fellow patient (complete report by 48 hour reduction is as soon as possible 24 hours). Another microorganism raise inspection produces the medical waste, according to this research experimental technique, may cause the traditional biochemical experiments which use originally to decide method produces the medical waste, by the approximately 6 kilograms reductions to the approximately 3.5 kilograms, reduced 42% infectious medical reject largely every day. These results indicate that the analysis of nucleic acid fragments is fast and simple, it provides a better method for identify the species of bacteria in order to improve the quality of therapy. |
