- 學位論文
利用UF-1000i尿沉渣分析儀結合SDS-NaOH溶液快速鑑別尿液檢體中革蘭氏陽性/陰性菌
Rapid Discrimination of Gram-Positive and Gram-Negative Bacteria in urine specimen by Using SDS-NaOH Solution and UF-1000i
摘要
泌尿道感染(urinary tract infection, UTI)是人類最常見的感染之一，臨床上會隨著年齡、性別及是否合併先天構造異常而有不同的臨床表徵。在診斷上可由尿液常規檢查中是否有膿尿、細菌、亞硝酸及白血球來評估，而細菌培養為診斷泌尿道感染的黃金準則。Sysmex UF-1000i全自動尿沉渣分析儀係利用流式細胞法針對細菌的核酸進行染色與偵測，可快速進行細菌定量及桿菌與非桿菌的初步分型但效率不高。另有文獻指出，利用革蘭氏陽性菌與革蘭氏陰性菌對SDS-NaOH溶液抗性的不同，可用來區分細菌型別。本研究目的在利用Sysmex UF-1000i的檢測結合SDS-NaOH對受測尿液進行處理，研討細菌型別之快速鑑別的可行性。方法為將含有細菌的尿液檢體，以SDS-NaOH溶液處理後以Sysmex UF-1000i分析，預測受測檢體中細菌的革蘭氏型別，最後與尿液培養比對兩者細菌型別的結果。結果發現經SDS-NaOH溶液在室溫下處理4分鐘後，可利用檢體中革蘭氏陰性菌幾乎會被完全溶解，而革蘭氏陽性菌則不易被溶解之特性來推斷細菌型別。此研究最終目的在於建立一個簡單快速的新方法，可以在常規尿液檢查中另外即時地提供細菌分型資訊給臨床醫師作為診斷與治療之參考。
並列摘要
Urinary tract infection (UTI) is one of the most common human infections. The symptoms with age, sex and whether to merge congenital structural anomalies have different performance. Currently, in the diagnosis by urine routine whether there is pyuria, bacteria, white blood cells and nitrite assessed by bacteria were cultured for further recognized as the gold standard. Sysmex UF-1000i, an automated urine analyzer, is by flow-based method for nucleic acid staining and bacteria count, it can quickly to discrimination of bacillus and non-bacillus. Literature indicated that the different resistance to SDS-NaOH solution of gram-positive bacteria and negative bacteria can be used to discrimination of bacterial gram-subtypes. In this study, we observed the change of bacteria count and bacteria scattergrams after urine specimens were treated with SDS-NaOH solution for 4 min at room temperature and then analyzed by Sysmex UF-1000i. We found that gram-negative bacteria could easily be lysed by direct addition of equal volumes of SDS-NaOH solution. In contrast, gram-positive bacteria could not be completely lysed by SDS-NaOH solution. This study aimed to describe simple and rapid methods that can discrimination of bacterial gram-subtypes in urine specimen by using Sysmex UF-1000i and SDS-NaOH solution. It also can easily applied in order to obtain additional information for clinical prescriptions from bacteriuria specimen with UTIs.