本文選題：實(shí)時(shí)熒光PCR + 核酸侵入反應(yīng)�。� 參考：《分析化學(xué)》2017年10期

【摘要】：通過檢測(cè)母體外周血中胎兒游離DNA(cffDNA)的SRY基因,確定胎兒性別,可評(píng)估胎兒性連鎖遺傳病的發(fā)病風(fēng)險(xiǎn),降低病兒出生率。本研究建立了高靈敏、高特異、閉管檢測(cè)不易污染的實(shí)時(shí)熒光PCR偶聯(lián)核酸侵入反應(yīng)方法用于SRY基因的檢測(cè)。通過優(yōu)化反應(yīng)體系中的檢測(cè)探針濃度、FEN1酶用量、Taq酶用量及預(yù)擴(kuò)增退火溫度,確定了最佳的反應(yīng)條件,即檢測(cè)探針濃度為250 nmol/L、FEN1酶用量為7.5 U、Taq酶用量為0.5 U、預(yù)擴(kuò)增退火溫度為67℃。在最佳反應(yīng)條件下,實(shí)現(xiàn)對(duì)含量低至4‰(4 copies/μL)的模擬樣本的檢測(cè),并成功檢測(cè)兩例孕期分別為9周和10周的臨床實(shí)際樣本。結(jié)果表明,所建立的方法可用于母體外周血cffDNA的SRY基因檢測(cè),為臨床開展基于SRY基因的無(wú)創(chuàng)產(chǎn)前診斷提供了新方法。
[Abstract]:By detecting the SRY gene of fetal free DNA in maternal peripheral blood and determining the fetal sex, we can evaluate the risk of fetal linked genetic disease and reduce the birth rate. In this study, a real-time fluorescent PCR coupled nucleic acid invasion method with high sensitivity, high specificity and low contamination was established for the detection of SRY gene. The optimum reaction conditions were determined by optimizing the probe concentration and the amount of FEN1 enzyme and Taq enzyme and the pre-amplification annealing temperature in the reaction system. The optimum reaction conditions were as follows: the detection probe concentration was 250 nmol / L FERN1 enzyme dosage 7.5 UTaq enzyme dosage was 0.5 UTaq enzyme dosage, the pre-amplification annealing temperature was 67 鈩，

本文編號(hào)：1819768