Abstract: Objectives　To explore the value of second generation sequencing in the diagnosis of central core disease (CCD). Methods　The clinical data and the results of detection of pathogenic genes by multiplex linked probe amplification and second generation sequencing in one CCD patient were retrospectively analyzed. Results　A 3-year-old girl presented slowly progressive walking difficulty, developmental retardation, facial myasthenia and congenital dislocation of hip joint. His parents had no similar clinical manifestations. Physical examination showed scoliosis, varying degrees of reduction in proximal muscle tension and muscle strength, muscle volume reduction and tendon reflex weakening, while pathological signs were negative. The patient's creatine kinase was slightly elevated and electromyography was normal. There was a novel missense mutation of c.14582G>A, p.Arg4861His (heterozygous) in the RYR1 gene detected by second generation sequencing method. The Alamut functional software predicted that the mutation could affect the structure and functions of the protein, and was highly related to CCD. Conclusion　The RYR1 gene mutation of c.14582G>A, p.Arg4861His is first reported in China. The second generation sequencing method can be used for the confirmation of diagnosis of suspected CCD.