改良固相BANA试验方法的建立

Establishment of an improved solid BANA test technique

目的:建立一种稳定、实用的改良固相BANA试验方法。方法:利用牙龈卟啉单胞菌、福赛氏类杆菌、螺旋体等牙周炎关键致病菌均能产生胰蛋白酶样酶(Trypsin-like enzyme,TLE),可使人工合成的酶底物—苯甲酰精氨酸萘酰胺(bengoyl-DL-arginine-naphthlamide,BANA)水解的原理,并以经典的液相法为"参照标准",在国外固相法的基础上进行了适当的改良,建立了改良固相BANA试验。结果:实验发现2mmol的BANA为改良固相BNA试验最适工作浓度,在55℃水箱孵育30min,可以检测到1.95×10-6Anson以上的胰蛋白酶,并具有良好的特异性和敏感性。结论:改良固相法与液相法一样具有较好的特异性和敏感性,但固相法更简单、快捷,为临床牙周病的检测提供了客观、简便的检测方法。

AIM： To establish a practical and stable solid BANA test technique. METHODS： P. gingivalis （P g）, B. forsythus （ Bf） and T. denticola （Td） can produce trypsin-like enzyme which is one of the important destructive factors for periodontal tissue. The hydrolyzing activity of trypsin - like enzyme on bengoyl - DL - arginine- naphthlamide （BANA） can be measured with routine liquid BANA test, a relative complicated techinque. An improved solid BANA test was established at the basis of foreign solid BANA test. RESULTS： 2mM BANA was the optimal substrate concentration. Over 1.95 ×10^-6 Anson unit trypsin was detected in hot water bath for 30 min at 55℃. The specificity and sensitivity were very high . CONCLUSION： The specificity and sensitivity of the improved solid BANA test were very high. The improved solide BANA test was quicker and more convenient than the routine liquid BANA test, and it might be an objective and useful index for diagnosis of clinical periodontitis.