Ritvaniemi P, Körkkö J, Bonaventure J, Vikkula M, Hyland J, Paassilta P, Kaitila I, Kääriäinen H, Sokolov B P, Hakala M
Jefferson Medical College of Thomas Jefferson University, Philadelphia, Pennsylvania, USA.
Arthritis Rheum. 1995 Jul;38(7):999-1004. doi: 10.1002/art.1780380717.
To use a recently developed procedure for analysis of blood leukocyte DNA to detect mutations in the gene for type II procollagen (COL2A1) in patients with cartilage diseases ranging from early-onset familial osteoarthritis (OA) to lethal chondrodysplasias.
The technique of denaturing gradient gel electrophoresis was used to scan polymerase chain reaction (PCR) products from 45 exons and exon-flanking sequences of the COL2A1 gene in more than 70 patients with cartilage diseases whose severity ranged from mild to lethal. PCR products with abnormal migrations were then sequenced.
Among the 3 patients with lethal hypochondrogenesis who were analyzed, all 3 were found to have a mutation in the COL2A1 gene. Among 17 patients with spondyloepiphyseal or spondyloepimetaphyseal dysplasia, 2 well-defined and 2 probable mutations were found. Among 15 patients with the Wagner-Stickler syndrome, 2 well-defined and 2 probable mutations were found. Among 45 patients with early-onset familial OA, 1 probable mutation was found.
Using the procedure developed for analysis of the COL2A1 gene, mutations were detected in > 20% of patients with chondrodysplasias and up to 2% of patients with early-onset familial OA. However, these percentages are only minimal estimates because all possible mutations in the gene cannot be detected with this procedure.
运用最近开发的血液白细胞DNA分析程序，检测从早发性家族性骨关节炎（OA）到致死性软骨发育不良等各类软骨疾病患者中Ⅱ型胶原蛋白（COL2A1）基因的突变情况。
采用变性梯度凝胶电泳技术，对70多名病情严重程度从轻度到致死性的软骨疾病患者的COL2A1基因45个外显子及外显子侧翼序列的聚合酶链反应（PCR）产物进行扫描。然后对迁移异常的PCR产物进行测序。
在分析的3例致死性低软骨生成患者中，全部3例均发现COL2A1基因突变。在17例脊椎骨骺或脊椎干骺端发育不良患者中，发现2个明确的和2个可能的突变。在15例瓦格纳-斯蒂克勒综合征患者中，发现2个明确的和2个可能的突变。在45例早发性家族性OA患者中，发现1个可能的突变。
运用为分析COL2A1基因而开发的程序，在超过20%的软骨发育不良患者以及高达2%的早发性家族性OA患者中检测到了突变。然而，这些百分比只是最低估计值，因为该程序无法检测到该基因的所有可能突变。
