结核DNA疫苗中卡那霉素残留的半定量检测法的建立及验证

Validation of Semi-quantitative Determination Method for Kanamycin Residue in Therapeutic Tuberculosis DNA Vaccine

目的用半定量法检测结核DNA疫苗中卡那霉素残留量。方法优化酶联免疫吸附法,验证该方法的准确度、中间精密度、专属性和耐用性。结果该方法能通过吸光度值的比较,判断供试品的含量,平均加样回收率为117%,准确度符合要求,其中间精密度符合要求。卡那霉素和其他3种相同浓度的抗生素(氨苄西林、庆大霉素、四环素)吸光度值相比,阳性率<1%,无交叉反应。当孵育温度和时间改变时,结果的RSD值均<20%。结论建立的方法有较好的准确性、中间精密度、专属性和耐用性,可用于结核DNA疫苗中卡那霉素残留量的检测。

OBJECTIVE To develop a semi-quantitative determination method for Kanamycin residue in therapeutic tuberculosis DNA vaccine. METHODS To verify the accuracy, intermediate precision, specificity, and durability of the method for optimizing the enzyme-linked immunosorbent assay(ELISA). RESULTS The ELISA method was accurately determined the content of the test sample by comparing the absorbance values. The average recovery rate of the sample was 117%. The accuracy and intermediate precision could meet the requirements. Compared with the other three antibiotics(ampicillin, gentamicin, tetracycline) at the same concentration, the positive rate of kanamycin was less than 1%,and had no cross-reaction. When the incubation temperature and incubation time changed, the results showed the RSD<20%. CONCLUSION The semi-quantitative method shows high accuracy, intermedia precision, specificity, and durability, which is suitable for detecting kanamycin residue in therapeutic tuberculosis DNA vaccine.