Development and Application of DNA Detection Kit for Deer Fetal Positive Control Sample
CUI Lin1, AI Jin-xia1*, DUAN Si-qi1, LIU Lin1, LI Ya-ru1, SHAO Bo-yu1, YANG Chang1, GAO Li-jun1, SUN Li-yuan1, LI Ming-cheng1,2, YUAN Guang-xin1
1. School of Medical Technology, Beihua University, Jilin 132013, China; 2. Innovation Center of DNA Fingerprint Detection Technology of Traditional Chinese Medicine, Jilin 132013, China
Abstract：OBJECTIVE To develop a deer fetus DNA positive control sample and assemble a test kit. METHODS Deer fetal DNA and its fake DNA were extracted, universal deer primers were designed and PCR amplified, species-specific target fragments were cloned, deer fetal DNA positive quality control products were constructed, and a DNA detection kit was assembled. The performance test of the kit was carried out and applied on commercially available deer fetus samples. RESULTS The sequencing comparison results were analyzed by NCBI. The accuracy rate was 100%. A clear band was found at 526 bp, which was consistent with the target fragment. The constructed sample can be used as a deer fetal positive control sample. The minimum detection limit of the sensitivity of the positive control was 10-1 ng·μL-1. After repeated freeze-thaw cycles, the stability of the PCR test results was proved. The test results of the commercially available deer fetuses and fakes using the kit were in line with expectations. CONCLUSION The constructed deer fetal positive control can provide more accurate and stable results for electrophoretic analysis and is suitable for the rapid identification of deer fetuses and fakes, which guarantees effective supervision of deer fetus quality.
崔琳, 艾金霞, 段思琪, 刘琳, 李亚茹, 邵博宇, 杨畅, 高丽君, 孙丽媛, 李明成, 苑广信. 鹿胎阳性质控品DNA检测试剂盒的开发与应用[J]. 中国药学杂志, 2021, 56(3): 232-237.
CUI Lin, AI Jin-xia, DUAN Si-qi, LIU Lin, LI Ya-ru, SHAO Bo-yu, YANG Chang, GAO Li-jun, SUN Li-yuan, LI Ming-cheng, YUAN Guang-xin. Development and Application of DNA Detection Kit for Deer Fetal Positive Control Sample. Chinese Pharmaceutical Journal, 2021, 56(3): 232-237.
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2021, Vol. 56 
