ABSTRACT
Objective@#To provide experimental evidence for genetic counseling and prenatal molecular diagnosis by analyzing the clinical characteristics and screening for pathogenic genes of a five-generation suspected multiple epiphyseal dysplasia (MED) family (17 patients).@*Methods@#The family members' medical history, general physical examination and hip joint X-ray examination were collected. Peripheral blood samples of the family members were collected and DNA were extracted from these samples. The exons of clinical genes from probands' DNA were sequenced by High throughput sequencing method. Next Gene software was used to compare and analyze the sequence and INGENUITY software was further used to annotate the mutations in order to find the pathogenic mutations in probands. The suspicious mutations were confirmed in pedigree members by PCR and Sanger sequencing.@*Results@#The family consisted of 5 generations and 38 members. Pedigree analysis was consistent with autosomal dominant inheritance. There were 17 patients in the family, and their clinical manifestations showed abnormal walking posture in childhood, pain in hip and knee joints, and typical pathological changes of epiphyseal dysplasia on X-ray. Cartilage oligomeric matrix protein (COMP) gene c.1153G>A (p.Asp385Asn) missense heterozygous mutation was screened in proband, which was genotypically and phenotypically segregated in the pedigree.@*Conclusion@#A missense mutation of the comp gene has been identified in a pedigree affected with MED which was the first reported in a big family. Our result is conducive to the further diagnosis and treatment and also provides a molecular basisfor the future prenatal diagnosis.
ABSTRACT
Objective To provide experimental evidence for genetic counseling and prenatal molecular diagnosis by analyzing the clinical characteristics and screening for pathogenic genes of a five-generation suspected multiple epiphyseal dysplasia (MED) family (17 patients).Methods The family members' medical history,general physical examination and hip joint X-ray examination were collected.Peripheral blood samples of the family members were collected and DNA were extracted from these samples.The exons of clinical genes from probands' DNA were sequenced by High throughput sequencing method.Next Gene software was used to compare and analyze the sequence and INGENUITY software was further used to annotate the mutations in order to find the pathogenic mutations in probands.The suspicious mutations were confirmed in pedigree members by PCR and Sanger sequencing.Results The family consisted of 5 generations and 38 members.Pedigree analysis was consistent with autosomal dominant inheritance.There were 17 patients in the family,and their clinical manifestations showed abnormal walking posture in childhood,pain in hip and knee joints,and typical pathological changes of epiphyseal dysplasia on X-ray.Cartilage oligomeric matrix protein (COMP) gene c.1153G ＞ A (p.Asp385Asn) missense heterozygous mutation was screened in proband,which was genotypically and phenotypically segregated in the pedigree.Conclusion A missense mutation of the comp gene has been identified in a pedigree affected with MED which was the first reported in a big family.Our result is conducive to the further diagnosis and treatment and also provides a molecular basisfor the future prenatal diagnosis.
ABSTRACT
Objective To identify the pathogens of 50 cases of fungal corneal ulceration by using semi-nested PCR amplification of ITS2 region.Methods Fifty isolates of fungal corneal ulceration and 3standard fungal strains cultures were collected and their DNAs were extracted.Their ITS2 regions were amplified by semi-nested PCR and sequenced.The results were compared with the nucleotide sequences in the NCBI GenBank.The pathogens of the fungi were identified and their distribution were analysed.Results The sequences results of the 3 standard fungal strains were consistent with the information in the GenBank.The pathological microorganisms of 50 cases of fungal corneal ulceration were:24 Fusarium (48％),including 17 Fusarium solani,6 Fusarium oxysporum and 1 Fusarium verticillioide; 10 Aspergillius (20％),including 5 Aspergillius flavus,3 Aspergillius sydowii and 2 Aspergillius nidulans; 6 Penicillium (12％),including 2 Penicillium citreo-viride,2 Penicillium multicolor and 2 Penicillium oxalicum ;5 Candida (10％),including 3 Candida albicans and 2 Candida parapsilosis; 3 Cladosporium (6％),including 2 Cladosporium herbarum and 1 case of Cladosporium cladosporioides ; 1 case of Neurospora crassa (2％) ;1 Alternaria alternata(2％).Conclusion Semi-nested PCR amplification of ITS2 region was proved to be a fast,simple and accurate method to identify pathogens of fungal corneal ulceration,and may be useful for personalized treatment and epidemiological investigation of local fungi.