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Minerva Biotecnologica 2020 March;32(1):1-5

DOI: 10.23736/S1120-4826.19.02567-9


language: English

TaqMAMA assay polymerase chain reaction real time for allelic discrimination of Macrophage receptor with collagenous structure rs1318645 polymorphism

Ilaria GALLIANO 1, 2, Valentina DAPRÀ 1, 3, Fernanda CIFERRI 2, Paola MONTANARI 1, 2, Cristina CALVI 1, 2, Carla ALLIAUDI 1, Francesco SAVINO 2, Massimiliano BERGALLO 1, 2, 3

1 Department of Public Health and Pediatrics, Medical School, University of Turin, Turin, Italy; 2 Department of Pediatrics, Regina Margherita Hospital, Città della Salute e della Scienza, Turin, Italy; 3 Biomole srl, Turin, Italy

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 ΔCt 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.

KEY WORDS: Real-time polymerase chain reaction; Bronchiolitis; Respiratory syncytial virus

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