鏈球菌的分類以及命名，在過去的15年以來一直在改變。這些改變多肇因於分子生物學方法的應用，如DNA-DNA hybridization及 16S核糖體核酸序列定序等。傳統的鑑定方法大多以型態特徵及生理生化反應做為基礎。本研究評估利用16S-23S核糖體核酸基因(rDNA)的內轉錄區(internal transcribed spacer, ITS)序列做為鑑定臨床常見鏈球菌之可能性。本實驗共使用52株(23種)參考菌株以及149株臨床菌株，利用PCR方法放大ITS序列並加以定序比對。結果顯示大多數的鏈球菌，其不同種間的相似度值介於0.33至0.94之間，相較於16S核糖體核酸基因序列，其不同種間序列相似度高達92到99％，利用ITS序列進行菌種鑑定是較好的選擇。本研究利用參考菌株序列與GenBank資料庫序列，整合架構鏈球菌ITS序列資料庫。根據實驗結果，此資料庫可準確鑑定臨床常見致病性鏈球菌，特別是較難鑑定的草綠色鏈球菌(viridans group streptococci, VGS)。本研究並應用這個資料庫中的序列，設計寡核甘酸微矩陣晶片 (oligonucleotide microarray chip), 能夠快速並正確鑑定草綠色鏈球菌種。該晶片設計可鑑定11種草綠色鏈球菌，共利用26株參考菌株及62株臨床分離株進行測試。比對後結果具有100％的靈敏度 (sensitivity)與特異性(specificity).綜合以上結論，ITS序列分析是一個正確、快速、且簡單的方法，提供鑑定臨床常見鏈球菌的另一種選擇。

The changes in taxonomy and nomenclature of Streptococcus have occurred in the past 15 years. These changes are the results of the application of DNA-DNA hybridization, 16S rRNA gene (rDNA) sequencing, and other molecular methods. Conventional identification methods of streptococci are mostly based on phenotypic and culture characteristics. In this study, 16S-23S rDNA internal transcribed spacer (ITS) regions of clinical significant streptococci were sequenced and evaluated for molecular identification of these bacteria. Fifty and two reference strains (23 species) and 149 clinical isolates were included in this approach. The ITS sequence similarity scores ranged from 0.33 to 0.94 among different streptococcal species. The ITS region seems to be a better choice for species identification as compared to the 16S rDNA sequences that have interspecies similarities in the range of 92-99 %. A database was established by using the ITS sequences obtained from reference strains and GenBank database. The database was found to be useful for identification of clinically important streptococci, especially for viridans group streptococci (VGS). Based on the database, an oligonucleotide microarray was developed for rapid identification of VGS. Twenty and six reference strains (11 species) and 62 clinical isolates of VGS were tested. A sensitivity of 100% and specificity of 100% were obtained. In conclusion, ITS sequence analysis could provide an alternative rapid and accurate method for species identification of clinically relevant streptococci.

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