CMP^(TM) MycoBeads分枝桿菌抗體磁珠應用於螢光染色及萋-尼兩氏抗酸性染色的效能

The Efficacy of CMP^(TM) MycoBeads for Used in the Fluorescence and the Ziehl-Neelsen Acid Fast Stain

羅晟展(Cheng-Chan Lo) ； 何耿德(Gen-Der Ho) ； 邱彥昕(Yen-Hsin Chiu) ； 黃玉君(Yu-June Huang) ； 楊士杰(Shih-Chien Yang) ； 謝賢修(Shian-Shiou Shie) ； 蔡岳廷(Yueh-Ting Tsai) ； 吳金洌

《檢驗及品保雜誌》 1卷2期 (2012/05) Pp. 59-68

https://doi.org/10.6573/JTQA.201205_1(2).02

摘要

本研究使用CMP^(TM) MycoBeads分枝桿菌磁珠（啟新生物科技有限公司，台灣）濃縮純化痰檢體中的分枝桿菌後製成之抹片操作螢光染色（fluorescence）及萋-尼兩氏抗酸性染色（Ziehl-Neelsen acid fast stain），將結果與直接塗抹片法、TB Beads（Microsens Medtech Ltd., London, UK）濃縮法和傳統離心濃縮法所製之抹片染色進行比較，評估鏡檢效能。實驗中使用9株不同來源之臨床分離結核分枝桿菌與1株結核分枝桿菌標準菌株（ATCC 25177），將10株菌分別以105 CFU混和痰液模擬臨床檢體狀態，得到抗酸菌的價數約為1+。檢體前處理過程如同一般臨床方法，但使用中性消化液（NALC-sodium citrate solution，不含2%NaOH），痰液化後，除進行直接塗抹片法外，再分別用離心法、TB beads及CMP^(TM) MycoBeads進行結核分枝桿菌濃縮純化製作抹片進行螢光染色及萋-尼兩氏抗酸性染色。以螢光顯微鏡（200×）或一般光學顯微鏡（1,000×）觀察染色結果並計數200個鏡場所觀察到的分枝桿菌總菌數。結果發現不論是螢光染色或萋-尼兩氏抗酸性染色，CMP^(TM) MycoBeads濃縮純化結核分枝桿菌的鏡檢偵測效能遠遠優於直接塗片之抗酸性染色法，並與離心法相當，且能顯著優於目前上市的磁珠產品TB Beads。

關鍵字

抗體磁珠 ； 濃縮 ； 抗酸性染色 ； 純化 ； 螢光染色

並列摘要

This study used the Mycobacterial magnetic beads (CMP^(TM) MycoBeads, Creative Microbiologicals, Ltd., Taiwan) and TB Beads (Microsens Medtech Ltd., London, UK) to concentrate Mycobacterium tuberculosis in simulated sputum specimens, and then applied to acid-fast staining. The total counts of mycobacteria (200 microscopic fields) were used to assess their performance. Nine M. tuberculosis clinical isolates and one M. tuberculosis reference strain (ATCC 25177) were used in the experiment. M. tuberculosis (10^5 CFU) was mixed respectively in sputum to simulate the clinical specimens. After the sputum specimens were digested by neutral NALC sodium citrate solution (without 2% NaOH), the bacteria in liquefied sputum were concentrated by centrifugation, TB beads or MycoBeads method, respectively. Pre-treated simulated sputum specimens were used to make smear for acid-fast stain (fluorescence and the Ziehl-Neelsen acid fast stain) to observe staining results and to count the total M. tuberculosis numbers in 200 fields. The results showed that regardless of the fluorescence stain or Ziehl- Neelsen acid-fast stain, the performance of MycoBeads was equally well comparable with centrifuge method and it's performance were significantly better than that of TB Beads method and direct smear method.

並列關鍵字

MycoBeads ； Fluorescence stain ； Ziehl-Neelsen acid-fast stain