单管5色荧光标记QF—PCR在常见染色体数目异常检测中的研究

Assessment of multiple quantitative fluorescence aneuploidies

目的评估单管5色荧光标记荧光定量PCR（QF—PCR）技术在常见染色体数目异常检测中的价值，以期优化产前诊断方案、缩短诊断周期。方法回顾性和前瞻性结合的临床对比研究。选取2013年8月至2015年9月于浙江大学医学院附属妇产科医院就诊的731例孕妇，超声指导下羊膜腔穿刺取羊水分别进行染色体核型分析及QF—PCR法短串联重复序列（STR）基因座检测。其中558例采用单盲法对常规染色体核型分析的剩余羊水标本作QF—PCR检测分析，173例采用双盲法将羊水标本同步作染色体核型分析和QF—PCR检测分析。结果采用单盲法分析的558例，用QF—PCR技术共检出2l三体5例、18三体2例、13三体1例、45X1例、47XXY1例、47XYY1例、47XXX1例和69XXX1例；采用双盲法173例，用QF—PCR技术共检出21三体1例、18三体1例。QF—PCR方法与染色体核型分析结果比较，阳性一致率达15／16，阴性一致率达100％（715／715），非嵌合体染色体数目异常检出率达15／15。结论单管5色荧光标记QF—PCR作为快速、准确、高通量、自动化产前诊断技术之一，实现其实验操作和结果判读的标准化和规范化，可用于诊断常见染色体（13，18，21，X，Y等）非整倍体异常，可作为染色体核型分析的重要补充，对于优化完善产前诊断体系，为孕妇提供更适宜的诊断方式有重要意义。

Objective To evaluate the effectiveness of muhiple quantitative fluorescence PCR （QF- PCR） as a rapid technique for prenatal diagnosis of common chromosome aneuploidies, in order to optimize the prenatal diagnosis and shorten the period of diagnosis. Methods Totally 731 amniotic fluid samples of pregnant subjects, who were referred to the Women＇s Hospital School of Medicine Zhejiang University during August 2013 and September 2015, were analyzed with conventional karyotype and the QF-PCR technique by short tandem repeat（STR） markers to detect chromosomes 13,18,21 ,X and Y aneuploidies. There were 558 samples detected by single blind method, 173 samples detected by double blind method. Results All of the 731 amniotic fluid samples were tested in this study by QF-PCR and the results were compared to the conventional eytogenetic analysis results of the same sample. Totally 558 samples with single blind method detected 5 trisomy 21, 2 trisomy 18, 1 trisomy 13, 1（45,X）, 1 （47,XXY）, 1 （47,XYY）, 1 （47,XXX） and 1 （69,XXX）, 173 samples with double blind method detected 1 trisomy 21 and 1 trisomy 18. The rapid QF-PCR assay was successful to detect all aneuploidies involving chromosomes 21, 18, 13, X and Y in prenatal diagnosis, which were verified by chromosome karyotype analysis. The results of QF-PCR method were compared with the results of chromosome karyotype analysis, the positive rate was 15/16, the negative rate was 100% （715/715）. Non chimeric chromosome abnormality detection rate was 15/15. Conclusions The multiple QF-PCR was a reliable method of detecting common chromosome aneuploidies for rapid prenatal diagnosis. As an important supplement of karyotype analysis, it was of great significance to optimize and improve the prenatal diagnosis system, and might provide more appropriate diagnostic methods for pregnant women.