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本文证实伤寒杆菌得以精确的遗传学技术制备的口服伤寒杆菌活菌苗具有表达耐酸应答(ATR)的能力,在pH5.0,厌氧条件下诱导ATR更为有效.作者将需氧培养物置37℃振摇16小时后按1:10稀释,再培养2小时,至培养物的波长600nm的吸光度(A600)为0.4.将厌氧培养物于37℃静置培养16小时,经标准化后使A600约为0.4.培养期间用缓冲液保持pH值不变.培养完毕后.在4℃下离心收集细菌.伤寒杆菌ATR评估方法是:上述细菌经生理盐水洗涤后在营养肉汤中(pH3.0)37℃培育90分钟,在0分钟和90分钟时分别取0.1ml悬浮菌液,以0.9ml 10mM的Tris-HCL(p3H7.5)中和并稀释,作活菌计数.
This article demonstrates that Salmonella typhimurium can be accurately gentrially produced viable Salmonella typhimurium oral vaccine capable of expressing an acid-fast response (ATR) and is more effective in inducing ATR under anaerobic conditions at pH 5.0. Shake 16 hours at 1:10 dilution, and then cultured for another 2 hours until the absorbance (A600) of the culture at a wavelength of 600nm was 0.4 The anaerobic culture was allowed to stand at 37 ° C for 16 hours, after standardization A600 About 0.4 during the incubation with the buffer to maintain the same pH After the culture was centrifuged to collect bacteria at 4 ° C. Salmonella typhi ATR assessment method is: the above bacteria after saline wash in nutrient broth (pH 3.0 ) At 37 ° C for 90 minutes. At 0 minutes and 90 minutes, 0.1 ml of the suspended bacterial liquid was taken, neutralized and diluted with 0.9 ml of 10 mM Tris-HCL (p3H7.5), and counted as viable cells.