本文選題：巢式PCR + 血漿游離胎兒DNA��； 參考：《中國(guó)輸血雜志》2016年08期

【摘要】：目的建立巢式PCR技術(shù)檢測(cè)RhD陰性孕婦血漿中游離胎兒DNA(cffDNA)的RHD基因型,以預(yù)測(cè)胎兒RhD血型。方法采用QIAamp DNA Blood Mini Kit提取32例RhD陰性孕婦血漿游離DNA,針對(duì)RHD外顯子7和10分別設(shè)計(jì)外側(cè)、內(nèi)側(cè)2組特異性引物,巢式PCR方法檢測(cè)孕婦血漿游離胎兒DNA的RHD型,測(cè)序驗(yàn)證PCR產(chǎn)物的序列特異性。結(jié)果孕婦血漿游離DNA經(jīng)巢式PCR擴(kuò)增后,有27例成功擴(kuò)增出RHD外顯子7、10特異性條帶,5例未檢測(cè)到RHD基因特異性擴(kuò)增,32例中有30例胎兒RHD型與出生后血型相符,檢測(cè)準(zhǔn)確率為93.1%。結(jié)論采用巢式PCR技術(shù)檢測(cè)RhD陰性孕婦血漿游離胎兒DNA來(lái)判定胎兒RHD型,具有良好的準(zhǔn)確性、敏感性和特異性,為RhD新生兒溶血病的早期診斷提供了一種新的、可靠的檢測(cè)手段。
[Abstract]:Objective to establish a nested PCR technique to detect the RHD genotypes of free fetal DNA (cffDNA) in the plasma of RhD negative pregnant women and to predict the fetal RhD blood group. Methods Plasma free DNA was extracted from 32 RhD-negative pregnant women by QIAamp DNA Blood Mini Kit. Two specific primers were designed for RHD exon 7 and 10, respectively. Nested PCR was used to detect the RHD type of free fetal DNA in maternal plasma. Sequence specificity of PCR products was verified by sequencing. Results in 27 cases of maternal plasma free DNA amplified by nested PCR, the RHD exon 710 specific band was successfully amplified in 5 cases. Of the 32 cases without RHD gene specific amplification, 30 cases of fetal RHD type corresponded to postnatal RHD blood type, and the detection accuracy was 93.1%. Conclusion Detection of free fetal DNA in plasma of RhD negative pregnant women by nested PCR has good accuracy, sensitivity and specificity, which provides a new and reliable method for the early diagnosis of RhD hemolytic disease.
【作者單位】： 陜西省血液中心;南方醫(yī)科大學(xué)南方醫(yī)院;深圳市血液中心;
【分類號(hào)】：R714.5;R440

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本文編號(hào)：2085861