评价多重连接探针扩增法诊断染色体22q11.2微缺失结果研究

目的 对多重连接探针扩增(multiplex ligation-dependent probe amplification,MLPA)诊断染色体22q11.2微缺失结果进行评价.方法 应用MLPA及荧光原位杂交(fluorescence in situ hybridization,FISH)两种方法分别检测了32份儿童(男16例,女16例;年龄1～13岁,平均3.6±3.1岁)血样本,其中16例为染色体22q11.2微缺失患儿组(阳性对照组),16名为体检正常儿童组(阴性对照组).采用灵敏度、特异度及Kappa分析来评估结果.结果 MLPA检测32份样本中,16例阳性对照样本均有染色体22q11.2微缺失,且缺失片段长度约3-Mb;16名对照样本中未发现22号染色体缺失.FISH证实16例22q11.2微缺失患儿均存在缺失,16名对照样本不存在缺失.因此,MLPA诊断染色体22q11.2微缺失的灵敏度及特异度高.结论 MLPA是一种快速、可靠、高通量及相对经济的诊断染色体22q11.2微缺失的有效方法,弥补了FISH技术的不足,可用于临床实验室快速诊断染色体22q11.2微缺失,具有较高的临床诊断价值.

Abstract：

Objective To evaluate multiplex ligation-dependent probe amplification (MLPA) assay detection in analysis of chromosome 22q11.2 microdeletion. Methods Between March 2008 and September 2009, thirty-two patients including 10 males and 16 females aged between years (3.6±3.1) were selected and evaluated by history, physical examination and medical records. Of these patients, sixteen patients who were previous diagnostic as 22q11.2 microdeletion were in positive control group, the other 16 healthy children were in negative control group. All the patients were detected by MLPA and fluorescence in situ hybridization (FISH) for the presence of a 22q1 1.2 microdeletion after informed consent. Diagnostic efficacy was assessed by sensitivity, specificity and Kappa analysis. Results We have applied the two assays of detection of chromosome 22q11.2 microdeletion in 32 patients. Sixteen patients in positive control group were found to have a 22q11. 2 deletion and, with the deletion size of 3-Mb. However, as expected,chromosome 22q11.2 deletion was not found in negative control group. The MLPA results were in good agreement with that by FISH. Therefore, MLPA has high sensitivity and specificity. Conclusion MLPA is a rapid, reliable, high-throughput and relatively economical alternative to FISH technology for the diagnosis of 22q11.2 microdeletion. It can provide reliable and helpful information for clinical diagnosis of 22q11.2 microdeletion syndrome.

Evaluation of detection and analysis of chromosome 22ql 1.2 microdeletion by multiple ligation-dependent probe amplification assay

Objective To evaluate multiplex ligation-dependent probe amplification (MLPA) assay detection in analysis of chromosome 22q11.2 microdeletion. Methods Between March 2008 and September 2009, thirty-two patients including 10 males and 16 females aged between years (3.6±3.1) were selected and evaluated by history, physical examination and medical records. Of these patients, sixteen patients who were previous diagnostic as 22q11.2 microdeletion were in positive control group, the other 16 healthy children were in negative control group. All the patients were detected by MLPA and fluorescence in situ hybridization (FISH) for the presence of a 22q1 1.2 microdeletion after informed consent. Diagnostic efficacy was assessed by sensitivity, specificity and Kappa analysis. Results We have applied the two assays of detection of chromosome 22q11.2 microdeletion in 32 patients. Sixteen patients in positive control group were found to have a 22q11. 2 deletion and, with the deletion size of 3-Mb. However, as expected,chromosome 22q11.2 deletion was not found in negative control group. The MLPA results were in good agreement with that by FISH. Therefore, MLPA has high sensitivity and specificity. Conclusion MLPA is a rapid, reliable, high-throughput and relatively economical alternative to FISH technology for the diagnosis of 22q11.2 microdeletion. It can provide reliable and helpful information for clinical diagnosis of 22q11.2 microdeletion syndrome.