新生兒表面蛋白A1基因多態(tài)性與支氣管肺發(fā)育不良相關(guān)性研究.pdf

1、華中科技大學(xué)碩士學(xué)位論文新生兒表面蛋白A1基因多態(tài)性與支氣管肺發(fā)育不良相關(guān)性研究姓名：張佳申請學(xué)位級別：碩士專業(yè)：兒科學(xué)指導(dǎo)教師：常立文2011-05華 中 科 技 大 學(xué) 碩 士 學(xué) 位 論 文 5Research on the correlation of gene polymorphism of surfactant protein A1 and bronchopulmonary dysplasia Abstract 【objec

2、tive】 To investigate the association between single-nucleotide polymorphisms(SNP) in the pulmonary surfactant Protein A1 gene and bronchopulmonary dysplasia 【Methods 】From Jul.2008 to Dec.2010 in Tongji Hospital，Tongji

3、Medical College，38 infants with BPD and 55 normal subjects in control were chose in the respective group.Whole-blood samples (2ml) from the umbilical cord were collected into plastic EDTA tubes and were prepared from blo

4、od using a routine phenol/trichloromethane method，then were stored at -20℃ until they were studied and genetype analysis was determined with the polymerse chain reaction restriction fragment length polymorphism(PCR-RFLP)

5、 method.To achieve specific and efficient PCR amplification ， we used two rounds of PCR amolificition.In the first round PCR，a gene-specific template was generated by a long PCR.Then the long PCR product was diluted 1:9

6、9 in dH2O and used as a template for the second PCR，which is nested-PCR.The PCR products(5µl) from the nested-PCR were digested with the appropriate enzyme and AA219 were water-bathed at 65℃ in two hours，while other

7、s were bathed at 37℃for one night.The allelic PCR fragments in the digested PCR products could be separated by electrophoresis and discriminated according to differences in length. 【Result】The first analysis was to ident

8、ify the type and frequency of each allele for both group，followed by an application of the Hardy-Weiberg trend test.The test yielded a unsignicant result.As the same mean，the samples in the group could represent the wh