The main aim of the present study is to analyze the impact of human rhinovirus (hRV) on patients with cystic fibrosis and their level of vitamin D3. To meet that aim, a statistical analysis is conducted. Forty samples were taken from patients’ pain from respiratory diseases, such as cystic fibrosis (CF) in Al-Noor teaching hospital in AL-Hilla town from December 2018 to February 2019. RNA illustrations were taken out from the nasopharyngeal swabs poised from patients thru cystic fibrosis (CF), succeeding the company’s guidelines. Based on the results, out of 40 patients with respiratory diseases, with cystic fibrosis (CF), a total of 6 (15%) human rhinovirus (hRV) isolates were recovered from nasopharyngeal swabs after they were subjected to reverse transcription-quantitative PCR (RT–qPCR). A total of 40 patients’ sera infected with cystic fibrosis (CF) and 40 control sera enrolled in this study were assessed for vitamin D3 (VD3) levels in their blood. Results demonstrate that VD3 concentrations were significantly lower in hRV positive than those negative and control (p < 0.05). It can be concluded that a serum focus of 25(OH)D is the recovered key of vitamin D prominence. It mirrors vitamin D bred cutaneous too that multiplied from food and