CAJ | 학술논문

基于三磷酸腺苷(ATP)光学反应原理,结合实验室自制的光学生物传感器,构建了一种快速检测卡介苗(BCG)活菌数的方法.选用加热裂解法提取BCG活菌内ATP,对BCG疫苗进行了检测.结果表明,BCG活菌浓度与相对发光强度(RLU)线性相关,相关系数为0.9908(P<0.01),其ATP含量与文献报道的结果处于同一数量级(10~(-18)mol/CFU).检测方法的相对标准偏差(RSD)为6.17%.检测样品仅需30μL,检测时间小于30 min.与国外商业化检测系统的测试结果线性相关,相关系数为0.9676(P<0.01).这种方法简便快速,在BCG疫苗及其他活菌疫苗质量控制方面具有广泛的应用前景.
기우삼린산선감(ATP)광학반응원리,결합실험실자제적광학생물전감기,구건료일충쾌속검측잡개묘(BCG)활균수적방법.선용가열렬해법제취BCG활균내ATP,대BCG역묘진행료검측.결과표명,BCG활균농도여상대발광강도(RLU)선성상관,상관계수위0.9908(P<0.01),기ATP함량여문헌보도적결과처우동일수량급(10~(-18)mol/CFU).검측방법적상대표준편차(RSD)위6.17%.검측양품부수30μL,검측시간소우30 min.여국외상업화검측계통적측시결과선성상관,상관계수위0.9676(P<0.01).저충방법간편쾌속,재BCG역묘급기타활균역묘질량공제방면구유엄범적응용전경.
Based on the principle of ATP bioluminescence reaction, a rapid method was developed for detecting the viability of BCG ( Bacillus Calmette Guerin) vaccine which combined homemade optical biosensor. ATP ( adenosine triphosphate) extraction was performed in boiling Tris-EDTA buffer in the experiments. In this study, a high correlation between the viable count of BCG and Relative luminescence intensity (RLU) was obtained with correlation coefficient of 0.9908 ( P < 0.01) according to the linearity of ATP standard, and a low Relative Standard Deviation (RSD) of 6.17%. The ATP content of BCG was 10~(-18) mol/CFU, consistent with the results in papers. The sample volume was 30 μL and the whole test for a sample was completed within 30 min. There was also a high cor-relation between homemade optical biosensor and the commercial system with correlation coefficient of 0.676(P < 0.01). With the characters of easy operation and fast responses, this method can be applied widely in routine quali-ty control to estimate viable count of BCG and other vaccines.