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５，１０亚甲基四氢叶酸还原酶（ＭＴＨＦＲ）是叶酸代谢的关键酶．为了试验肌肉介导外源基因人ＭＴＨＦＲ（ｈＭＴＨＦＲ）制备抗体和建立ＭＴＨＦＲ免疫检测的可能性，构建了ＭＴＨＦＲ基因真核表达载体（ｐｃＤＮＡ３／ＭＴＨＦＲ）；通过基因缝线法将携带ｐｃＤＮＡ３／ＭＴＨＦＲ的质粒，缝合于预先注射再生剂（丁哌卡因）的肌肉内．２个月后分离血清，所得抗体应用Ｗｅｓｔｅｒｎｂｌｏｔ，ＥＬＩＳＡ和胎肝免疫组织化学染色进行免疫鉴定．胎肝免疫组织化学显示，在肝小梁细胞浆中具有大量ＭＴＨＦＲ阳性反应颗粒；Ｗｅｓｔｅｒｎｂｌｏｔ有ＭＴＨＦＲ抗体与其抗原特异的褐色条带，分子量约为３７ｋＤ；ＥＬＩＳＡ分析表明，３种不同浓度的抗体与不同剂量的抗原反应具有剂效关系，最适抗体滴度（ＥＤ５０）为１∶４００。以上结果说明肌肉介导外源基因是获得抗体的一种简单、快捷的方法．该抗体可用于ＭＴＨＦＲ的免疫检测和有关的叶酸代谢研究工作．
5,10 methylene tetrahydrofolate reductase (MTHFR) is the key enzyme in folic acid metabolism. MTHFR eukaryotic expression vector (pcDNA3 / MTHFR) was constructed in order to test the muscle-mediated production of antibodies against human MTHFR (hMTHFR) and to establish the MTHFR immunoassay. Plasmids carrying pcDNA3 / MTHFR Sutured into the muscle of a pre-injected regenerative agent (bupivacaine). After 2 months, the serum was separated and the antibodies were identified by Western blot, ELISA and fetal liver immunohistochemical staining. Fetal liver immunohistochemistry showed a large number of MTHFR-positive reaction particles in the trabecular cell cytoplasm; Westernblot with MTHFR antibody and antigen-specific brown band, molecular weight of about 37kD; ELISA analysis showed that three different concentrations of antibodies and different Dose of the antigen response with a dose-effect relationship, the optimal antibody titer (ED50) of 1: 400. The above results indicate that muscle-mediated exogenous gene is a simple and quick method to obtain antibodies. The antibody can be used for MTHFR immune detection and related folate metabolism research.