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评估 TaqMan Jr 基因分型法快速预测抗-Jr 抗体存在的方法。

Evaluating the TaqMan Jr-Genotyping Method for Rapidly Predicting the Presence of Anti-Jr Antibodies.

机构信息

Department of Laboratory Medicine, Severance Hospital, Yonsei University College of Medicine, Seoul, Korea.

出版信息

Ann Lab Med. 2024 Sep 1;44(5):418-425. doi: 10.3343/alm.2023.0325. Epub 2024 Feb 20.

Abstract

BACKGROUND

The Jr antigen is a high-prevalence red blood cell (RBC) antigen. Reports on cases of fatal hemolytic disease of the fetus and newborn and acute hemolytic transfusion reactions suggest that antibodies against Jr (anti-Jr) have potential clinical significance. Identifying anti-Jr is challenging owing to a lack of commercially available antisera. We developed an alternative approach to rapidly predict the presence of anti-Jr using the TaqMan single-nucleotide polymorphism (SNP)-genotyping method.

METHODS

Residual peripheral blood samples from 10 patients suspected of having the anti-Jr were collected. Two samples with confirmed Jr(a-) RBCs and anti-Jr were used to validate the TaqMan genotyping assay by comparing the genotyping results with direct sequencing. The accuracy of the assay in predicting the presence of anti-Jr was verified through crossmatching with in-house Jr(a-) O+ RBCs.

RESULTS

The TaqMan-genotyping method was validated with two Jr(a-) RBC- and anti-Jr-confirmed samples that showed concordant Jr genotyping and direct sequencing results. Jr genotyping for the remaining samples and crossmatching the serum samples with inhouse Jr(a-) O+ RBCs showed consistent results.

CONCLUSIONS

We validated a rapid, simple, accurate, and cost-effective method for predicting the presence of anti-Jr using a TaqMan-based SNP-genotyping assay. Implementing this method in routine practice in clinical laboratories will assist in solving difficult problems regarding alloantibodies to high-prevalence RBC antigens and ultimately aid in providing safe and timely transfusions and proper patient care.

摘要

背景

Jr 抗原是一种高频率出现的红细胞(RBC)抗原。有关胎儿和新生儿溶血病和急性溶血性输血反应的报告表明,针对 Jr 的抗体(抗-Jr)具有潜在的临床意义。由于缺乏商业上可获得的抗血清,因此鉴定抗-Jr 具有挑战性。我们开发了一种替代方法,使用 TaqMan 单核苷酸多态性(SNP)-基因分型方法来快速预测抗-Jr 的存在。

方法

收集了 10 例疑似存在抗-Jr 的患者的残留外周血样本。使用两种经确认 Jr(a-) RBC 和抗-Jr 的样本,通过将基因分型结果与直接测序进行比较,验证 TaqMan 基因分型检测的准确性。通过与内部 Jr(a-) O+ RBC 进行交叉配血,验证了该检测方法预测抗-Jr 存在的准确性。

结果

TaqMan 基因分型方法通过两种经确认 Jr(a-) RBC 和抗-Jr 的样本进行了验证,这两种样本的 Jr 基因分型和直接测序结果一致。对其余样本进行 Jr 基因分型,并将血清样本与内部 Jr(a-) O+ RBC 进行交叉配血,结果一致。

结论

我们验证了一种使用基于 TaqMan 的 SNP 基因分型检测快速、简单、准确且具有成本效益的方法来预测抗-Jr 的存在。在临床实验室常规实践中实施这种方法将有助于解决针对高频率 RBC 抗原的同种抗体的难题,并最终有助于提供安全及时的输血和适当的患者护理。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c146/11169768/ae0f0bb338e5/alm-44-5-418-f1.jpg

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