Mutation screening in 48 non-syndromic short stature Chinese patients
Forty-eight non-syndromic short stature Chinese patients were recruited in Shanghai Children’s Medical Center. Their age, gender and height information are included in Additional file 1. The inclusion criteria were individuals with height below 3rd percentile without a clinical diagnosis of intellectual disability or developmental delay. All information was obtained with appropriate consent based on requirements of Shanghai Children’s Medical Center【SCMC-IRB-K2013007】.Subjects were randomly selected in non-syndromic short stature patients. Since it is unclear yet if ARID1B affects hormone-related pathways, we did not use hormonal status as a criteria for subject selection. Genomic DNA was extracted from peripheral blood of all participants using QIAamp Blood DNA Mini kit®. Mutation screening for all coding regions of ARID1B were done by Polymerase chain reaction (PCR) amplification followed by Sanger sequencing. Sequence variants were evaluated with mutation surveyor (Soft Genetics, State College, PA) and their potential functional impact was predicted using insilico prediction programs including SIFT [5], Polyphen2 [6], Condel [7] and Align-GVGD [8]. Paternity tests were performed with short tandem repeat (STR) markers for the two probands with de novo variants (using AmpFLSTR® Identifiler® PCR amplification kit). The study was reviewed and approved by the SCMC ethical committee and all participants or their parents signed an informed consent form. In addition, we compared the variant frequency in the ARID1B coding regions detected by exome sequencing to that of 494 normal Chinese controls. The normal Chinese controls were age and gender matching Chinese individuals of normal height, weight and were recruited from multiple geographic areas for an effort to create a common sequence variants database of normal Chinese children.