BACKGROUND: Macrophage receptor with collagenous structure (MARCO), is an innate immunity scavenger receptor as possible gene candidate to respiratory syncytial virus (RSV) disease susceptibility. We developed a TaqMAMA real time PCR for detection of rs1318645 MARCO SNP in infant with bronchiolitis infection. METHODS: We enrolled 59 healthy term infants and 60 bronchiolitis. TaqMAMA assays has been designed for discrimination of the MARCO alleles. Specificity has been assessed evaluating ACt of a homozygous DNA sample intended as the difference between the specific and aspecific amplifications. RESULTS: No ΔCt value was obtained for the two primer and probe conditions tested. Coefficient of variation (CV) for MARCOc and -g was under 1% and our group in previously TaqMAMA development assay. All 119 samples were tested with 70/119 (58.8%) homozygotes (30 CC and 40 GG) and 49/119 (41.2%) heterozygotes (CG). Diverse papers in literature displayed an association between the presence of MARCO rs1318645 polymorphism and RSV disease susceptibility. CONCLUSIONS: We identified CC homozygotes to predispose to RSV infection. The method here described needful negligible development time, return data that are straightforward to analyze and had a high possibility of success.
TaqMAMA assay polymerase chain reaction real time for allelic discrimination of Macrophage receptor with collagenous structure rs1318645 polymorphism
Galliano I.;Dapra V.;Ciferri F.;Calvi C.;Alliaudi C.;Bergallo M.
2020-01-01
Abstract
BACKGROUND: Macrophage receptor with collagenous structure (MARCO), is an innate immunity scavenger receptor as possible gene candidate to respiratory syncytial virus (RSV) disease susceptibility. We developed a TaqMAMA real time PCR for detection of rs1318645 MARCO SNP in infant with bronchiolitis infection. METHODS: We enrolled 59 healthy term infants and 60 bronchiolitis. TaqMAMA assays has been designed for discrimination of the MARCO alleles. Specificity has been assessed evaluating ACt of a homozygous DNA sample intended as the difference between the specific and aspecific amplifications. RESULTS: No ΔCt value was obtained for the two primer and probe conditions tested. Coefficient of variation (CV) for MARCOc and -g was under 1% and our group in previously TaqMAMA development assay. All 119 samples were tested with 70/119 (58.8%) homozygotes (30 CC and 40 GG) and 49/119 (41.2%) heterozygotes (CG). Diverse papers in literature displayed an association between the presence of MARCO rs1318645 polymorphism and RSV disease susceptibility. CONCLUSIONS: We identified CC homozygotes to predispose to RSV infection. The method here described needful negligible development time, return data that are straightforward to analyze and had a high possibility of success.
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