KL-BoBs联合QF-PCR技术对流产组织检测的临床应用评估

Clinical Application Assessment of KaryoLite BoBs Combined with QF-PCR in the Detection of Products of Conception

目的通过用核型细菌人工染色体标记-微球鉴别/分离法(KL-BoBs)对早孕期流产组织进行遗传学检测,评估KL-BoBs联合荧光定量PCR技术(QF-PCR)对流产组织进行遗传学检测的准确性。方法收集2016年5~8月在四川大学华西第二医院产前诊断中心进行染色体微阵列分析(CMA)检测的81例早孕期流产组织样本(61例胎盘绒毛组织,19例胎儿肌肉组织,1例胎儿肝脏组织),用KL-BoBs及QF-PCR技术对样本进行检测,将检测结果与CMA检测结果进行对比,评估KL-BoBs与QF-PCR联合检测的准确性。结果在81例流产组织样本中,70例样本经KL-BoBs检测的结果与其CMA检测结果一致,包括36例正常核型、34例异常核型(非整倍体);KL-BoBs不能检出胎儿三倍体(结果显示为2例正常核型和5例非整倍体),CMA和QF-PCR均检出;KLBoBs不能检出较小片段的拷贝数变异。CMA检测出4例样本拷贝数变异。KL-BoBs联合QF-PCR方法与CMA阳性诊断的符合率为91.1%(41/45),阴性诊断的符合率为100%(36/36)。KL-BoBs的检测准确率为86.4%(70/81),假阳性率为0%,假阴性率为13.3%(6/45),如果同时进行KL-BoBs和QF-PCR检测,可将准确率提高到95.1%(77/81)。结论 KL-BoBs联合QF-PCR检测早孕期流产组织准确率高,可作为流产组织染色体异常的一线检测方法。

Objective To assess the accuracy and discuss the feasibility of KaryoLite bacterial artificial chromosome on beads （KL-BoBs） and quantitative fluorescent polymerase chain reaction （QF-PCR） in genetic testing of products of conception （POC） by comparing with the chromosomal microarray analysis （CMA） test results. Methods Eighty-one cases of abortion samples were collected in the prenatal diagnosis center of West China Second University Hospital in Sichuan University from May to August 2016, including 61 cases of placenta tissues, 19 cases of fetal muscle tissues and 1 case of fetal liver tissue. KL-BoBs and QF-PCR were used to detect the samples. The results were compared with those of CMA test to evaluate the accuracy of KL-BoBs and QF-PCR. Results Of the 81 POC samples, the results of 70 samples tested by KL-BoBs was consistent with that of CMA. Among them, 36 cases were normal karyotype and 34 cases were abnormal karyotypes （aneuploidy）. Triploid could not been detected by KL-BoBs （the results were shown 2 cases were normal karyotype and 5 cases were aneuploidy）, whereas CMA and QF-PCR could be detected. Copy number variation of small segments could not been detected by KL-Bobs. Four cases of copy number variationwere detected by CMA. Compared with CMA, the positive coincident rate of KL-BoBs combined with QF-PCR was 91.1% （41/45）, the negative coincidence rate was 100% （36/36）. The accuracy rate of KL-BoBs was 86. 4% （70/81）, the false positive was 0% and the false negative was 13.3% （6/45）. Whereas both KL-BoBs and QF-PCR were simultaneously detected, the accuracy rate would be improved to 95.1% （77/81）. Conclusion The accuracy rate of KL-BoBs combined with QF-PCR was high for testing early pregnancy abortion tissue. It can be used as a first-tier test.