期刊文献+

免疫磁珠法与差异裂解法检验混合样本的效果观察 被引量:1

Comparative study of magnetic beads method and differential lysis method for the separation and detection of mixed cell samples
原文传递
导出
摘要 目的分析应用免疫磁珠分离技术和差异裂解法对混合样本的DNA检验效果,评价其在法医学应用中的价值。方法制备含有不同数量精细胞与阴道上皮细胞混合悬液和植绒混合斑拭子,分别采用MOSPD3抗体、SPAG8抗体免疫磁珠法及差异裂解法对样本进行检验,观察分析不同比例的混合细胞悬液,以及不同保存时间植绒混合斑拭子DNA分型的正确率。结果当混合细胞悬液内精细胞与阴道上皮细胞比例约1∶10时,免疫磁珠法的正确率明显高于差异裂解法(P<0.01);当比例等于或高于1∶1时,两种方法正确率的差异无显著性意义(P>0.05)。植绒拭子混合斑的免疫磁珠法正确率明显低于差异裂解法(P<0.01);随混合斑保存时间的延长,差异裂解法正确率无明显下降(P>0.05)。结论免疫磁珠法适用于新鲜混合样本中精细胞的分离检验,且在精细胞含量较低时优势更明显,差异裂解法较适合陈旧混合斑的分离检验。 Objective Results of antibody-magnetic beads method and differential lysis method for the separation of sperm in mixed cell samples were compared, to explore the potential feasibility in forensic practice. Methods Three different concentration mixed cell suspensions and simulated stains of sperm and vaginal epithelial cell was prepared. All the samples of mixed cell suspensions and simulated mixed stains were detected by antibody-magnetic beads method and differential lysis method respectively. The number of STR loci and the correct detection rate were calculated and compared. Results The correct detection rate of antibody-magnetic beads method was higher than differential lysis method when the number of sperm cells and vaginal epithelial cell was approximately 1 : 10 ( P 〈 0. 01 ). While there was no significant difference for the two methods when the number of sperm cells and vaginal epithelial cell was equal to or more than 1 : 1 (P 〉 0.05 ). The correct detection rate of antibody-magnetic beads method was lower than differential lysis method for the samples of simulated mixed stains in different storage time groups (P 〈 0. 01 ). While there was no significant difference for the differential lysis method with the samples preservation time lasting (P 〉 0. 05 ). Conclusion Antibody-magnetic beads method was effective for the fresh mixed samples especially when contains few sperm cells. Differential lysis method was more suitable for the old mixed samples.
出处 《中国法医学杂志》 CSCD 2015年第3期249-251,255,共4页 Chinese Journal of Forensic Medicine
基金 山东省自然科学基金资助项目(ZR2014HQ018) 山东省高校证据鉴识重点实验室(山东政法学院)开放课题(2014-01) 重庆市公安局科技攻关项目(2014-10)
关键词 法医物证学 免疫磁珠 混合斑 精子 DNA forensic biological evidence immunomagnetic beads mixed stain sperm cell DNA
  • 相关文献

参考文献11

  • 1李学博,王清山,宁淑华,王业全,封宇,缪元颖,李红卫.利用精子特异性抗体SPAG8分离混合细胞体系中的精子[J].第三军医大学学报,2014,36(12):1322-1326. 被引量:2
  • 2Li X,Wang Q,Feng Y,et al.Magnetic bead-based separation of sperm from buccal epithelial cells using a monoclonal antibody against mospd3[J].Int J Legal Med.2014,128(6):905-911. 被引量:1
  • 3王清山,宁淑华,李学博,李红卫,林子清.应用免疫磁珠分离混合斑中精子[J].中国法医学杂志,2013,28(4):317-319. 被引量:3
  • 4Yoshida K,Sekiguchi K,Mizuno N,et al.The modified method of two-step differential extraction of sperm and vaginal epithelial cell DNA from vaginal fluid mixed with semen[J].Forensic Sci Int,1995,72(1):25-33. 被引量:1
  • 5Schulz M M,Buschner M G,Leidig R,et al.A new approach to the investigation of sexual offenses-cytoskeleton analysis reveals the origin of cells found on forensic swabs[J].J Forensic Sci,2010,55(2):492-498. 被引量:1
  • 6Vandewoestyne M,Van Nieuwerburgh F,Van Hoofstat D,et al.Evaluation of three DNA extraction protocols for forensic str typing after laser capture microdissection[J].Forensic Sci Int Genet,2012,6(2):258-262. 被引量:1
  • 7Horsman KM,Barker SL,Ferrance JP,et al.Separation of sperm and epithelial cells in a microfabricated device:Potential application to forensic analysis of sexual assault evidence[J].Anal Chem,2005,77(3):742-749. 被引量:1
  • 8赵兴春,姜伯玮.精子细胞定向捕获与分离技术初步研究[J].刑事技术,2012,37(1):14-17. 被引量:13
  • 9Wascher R A,Huynh K T,Giuliano A E,et al.Stanniocalcin-1a novel molecular blood and bone marrow marker for human breast cancer[J].Clin Cancer Res,2003,9(4):1427-1435. 被引量:1
  • 10Feng H L,Han Y B,Sparks A E,et al.Characterization of human sperm antigens reacting with anti-sperm antibodies from an infertile female patient's serum[J].J Androl,2008,29(4):440-448. 被引量:1

二级参考文献31

  • 1袁自闯,赵凯,李海燕,孙宏钰,欧桂生.Differex^(TM)系统在混合斑精子DNA分离提取中的应用[J].中国法医学杂志,2006,21(2):99-99. 被引量:6
  • 2郑会芬,董研,尚宝良.混合斑中精子DNA抽提方法的改进[J].中国司法鉴定,2006(3):35-35. 被引量:5
  • 3李鑫,胡兰,郭红玲,刘志芳,凃政,刘开会,米瑞华.显微操作法提取混合斑中精子细胞方法的探讨[J].中国法医学杂志,2006,21(5):263-265. 被引量:19
  • 4袁自闯,李海燕,赵凯,宁忠,唐振亚,邝玉斌,欧桂生.3种分离提取混合斑精子DNA的方法比较[J].中国法医学杂志,2007,22(5):325-326. 被引量:10
  • 5Chen J, Kobilinsky L, Wolosin D, et al. A physical method for separating spermatozoa from epithelial cell in sexual assault evidence[J]. J Forensic Sci,1998,43( 1): 114. 被引量:1
  • 6Elliott K, HillD S, lambertc C, et al. Use of lasermicrodissection greatly impoves the recovery of DNA from sperm on microscope slides [ J ]. Forensic Sci Int, 2003, 137(1) :28-36. 被引量:1
  • 7Gusmaol, Gonzalez-Neira A, Pestoni C, et al. Robustness of the Y STRs DYS19, DYS389 I and I1, DYS390 and DYS393: optimization of a PCR penaplex [ J ]. Forensic Sci Int, 1999,106 : 163. 被引量:1
  • 8Lei Jiahui, Guan Fei, Xu Hong, et al. Application of an immunomagnetic bead ELISA based on IgY for detection of circulating antigen in urine of mice infected with Schistosoa japonicum [ J ]. Vet Parasitol. 2012,187 : 196-202. 被引量:1
  • 9Liu D Y, Marticl M, Clarke G N, et al. An anti-actin monoclonal antibody inhibits the zonapellucida-induced acrosome reaction and hyperactivmotility of human sperm [J]. Mol Hum Reprod. 2002, 8(1):37-47. 被引量:1
  • 10Marshall P. Optimization of spermatozoa capture during the differential extraction process for STR typing with the potential for automation. Master's thesis. University of North Texas. 被引量:1

共引文献14

同被引文献22

引证文献1

二级引证文献5

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部