Research On The Correlation Of Gene Polymorphism Of Surfactant Protein A1 And Bronchopulmonary Dysplasia

【objective】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 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 blood 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) 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 1:99 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 others 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 identify 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 whole population well. In the BPD group,the distribution of AA50 loucs genetype(P=0.038) and G/C allele(P=0.012)differed significantly with control cases.The frequencies of AA219, AA50 and AA19 genetypes and alleles were not different between the BPD infants and normal controls.The frequency of the C/Cgenetype in AA50 was significanty lower in the infants with BPD than in the normal controls.【Conclusion】The results indicate that a gene association exists between SPA1 gene and BPD. In BPD,possession of the C/C genetype in the AA50 would reduce the the risk of BPD...