Cheng Lin, Feng Chun, Wu Ke, Wang Zixiang, Zhu Wentao, Geng Ting, Li Naiqi, Kang Jiawei, Duan Jie, Li Jin, Shao Yuqi, Yang Saisai, Peng Jianhong, Cai Bo, Ma Jianhong, Zhao Xingzhong, Zhang Yuanzhen
Department of Obstetrics, Zhongnan Hospital of Wuhan University, Wuhan, 430071, China; Hubei Clinical Research Center for Prenatal Diagnosis and Birth Health, Wuhan, 430071, China.
Department of Gynecology, Maternal and Child Health Hospital of Hubei Province, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430070, China.
Anal Chim Acta. 2025 Mar 15;1343:343690. doi: 10.1016/j.aca.2025.343690. Epub 2025 Jan 20.
Fetal monogenic disorders cause gestation anomalies, birth defects or infant mortality. Implementing non-invasive prenatal testing (NIPT) of these diseases benefits timely intervention while avoids high risk of inducing maternal complications and fetal injuries. However, fetal materials used for current NIPT (e.g., blood-borne cell-free fetal DNA (cffDNA)) are inevitably interfered by tremendous maternal interference in peripheral blood, leading to difficulty of exactly analyzing fetal monogenic mutations. Fetal nucleated red blood cells (FNRBCs) in maternal peripheral blood possess the entire fetal genome, which is encapsulated inside the cells and is away from maternal interference. Therefore, FNRBCs are promising and superior for NIPT of fetal genetic disorders after they are highly purified from huge maternal blood cells and subsequently analyzed.
We synthesized multi-functional magnetic nanoparticles (MMNs) by coating erythrocyte-leukocyte hybrid membrane on FeO magnetic nanoparticles and grafting specific antibody (anti-CD147) to highly purify FNRBCs from maternal peripheral blood. Fetal origin of all the isolated cells were confirmed by short tandem repeat (STR) to validate their sufficient purity for downstream analysis. Potential maternal factors that would influence FNRBC counts were evaluated based on clinical samples. Utilizing Sanger sequencing and droplet digital PCR (ddPCR), FNRBCs were analyzed to non-invasively assess the risk of five kinds of fetal monogenic disorders, which was challenging for current NIPT.
It is anticipated that this MMN-based strategy will create possibilities for overcoming limitations of NIPT when detecting fetal monogenic disorders nowadays.
胎儿单基因疾病会导致妊娠异常、出生缺陷或婴儿死亡。对这些疾病实施无创产前检测（NIPT）有利于及时干预，同时避免诱发母体并发症和胎儿损伤的高风险。然而，目前NIPT所使用的胎儿材料（如血源游离胎儿DNA（cffDNA））不可避免地受到外周血中大量母体干扰的影响，导致准确分析胎儿单基因突变存在困难。母体外周血中的胎儿有核红细胞（FNRBCs）拥有完整的胎儿基因组，该基因组封装在细胞内，不受母体干扰。因此，从大量母体血细胞中高度纯化FNRBCs并随后进行分析后，它们在胎儿遗传疾病的NIPT中具有广阔前景且更为优越。
我们通过在FeO磁性纳米颗粒上包覆红细胞-白细胞混合膜并接枝特异性抗体（抗CD147）来合成多功能磁性纳米颗粒（MMNs），以从母体外周血中高度纯化FNRBCs。通过短串联重复序列（STR）确认所有分离细胞的胎儿来源，以验证其纯度足以进行下游分析。基于临床样本评估了可能影响FNRBC计数的潜在母体因素。利用桑格测序和液滴数字PCR（ddPCR）对FNRBCs进行分析，以无创评估五种胎儿单基因疾病的风险，这对当前的NIPT来说具有挑战性。
预计这种基于MMN的策略将为克服当前NIPT在检测胎儿单基因疾病时的局限性创造可能性。
