Novel, non-radioactive, simple and multiplex PCR-cRFLP methods for genotyping human SP-A and SP-D marker alleles

We have previously identified an allele of the human SP-A2 gene that occurs with greater frequency in an RDS population [12]. Because of the importanc e of SP-A in normal lung function and its newly emerging role in innate hos t defense and regulation of inflammatory processes, we wish to better chara cterize genotypes of both SP-A1 and SP-A2 genes. It has been determined tha t SP-D shares similar roles in immune response. Therefore, in this report w e 1) describe a novel, non radioactive PCR based-cRFLP method for genotypin g both SP-A and SP-D; 2) describe two previously unpublished biallelic poly morphisms within the SP-D gene; 3) present the partial sequence of one new SP-A1 allele (6A(14)) and describe other new SP-A1 and SP-A2 alleles; and 4 ) describe additional methodologies for SP-A genotype assessment. The abili ty to more accurately and efficiently genotype samples from individuals wit h various pulmonary diseases will facilitate population and family based as sociation studies. Genetic polymorphisms may be identified that partially e xplain individual disease susceptibility and/or treatment effectiveness.