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

作　　者：李婷 谭奇凤 杨丽菲 孔雯雯 倪世明 黄明 梁艳 吴雪琼 伊君梅 田园 LI Ting;TAN Qifeng;YANG Lifei;KONG Wen wen;NI Shiming;HUANG Ming;LIANG Yan;WU Xueqiong;YI Junmei;TIAN Yuan(Guangzhou Baiyunshan Baidi Bio-Technology Co.,Ltd.,Guangzhou,Guangdong 511495,China;Tuberculosis Key Laboratory,the Sth Medical Center,PLA General Hospital,Beijing 100091,China;Gu'an Dingtai Haigui Biotechnology Co.,Ltd.,Langfang,Hebei 065500,China)

机构地区：[1]广州白云山拜迪生物医药有限公司,广东广州511495 [2]中国人民解放军总医院第八医学中心结核病重点实验室,北京100091 [3]固安鼎泰海规生物科技有限公司,河北廊坊065500

出　　处：《今日药学》2021年第9期678-681,共4页Pharmacy Today

基　　金：国家科技重大专项(2018ZX10731301-005-003)。

摘　　要：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.

关 键 词：抗生素残留 半定量法 酶联免疫吸附法 验证 

分 类 号：R917[医药卫生—药物分析学]