Fully genotyping and screening of clinically important blood‐group antigens by MALDI TOF mass spectrometry

Abstract

Several molecular biology methods are available for high‐throughput blood typing. In this study, we aimed to build a high‐throughput blood‐group genetic screening system for high‐frequency blood‐group antigen‐negative rare‐blood groups in donors and patients. The amplification primers for all blood‐type gene fragments involving the selected alleles were designed for detection. Single‐base extend primers were also designed based on specific loci. DNA fragments were detected by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MS) for the last nucleotide identification of amplification products in the extend step. The accuracy was verified by known samples. Thirty‐six random samples were detected by serological tests and sequencing to verify the system stability. After verification, according to the collected known rare‐blood‐type samples, all the alleles designed to be detected matched with the validated single‐nucleotide polymorphisms. The verification tests showed that all genotyping results of the random samples were in accordance with the findings of serotyping and sequencing. Then, 1258 random donor samples were screened by the built typing system after the verification. Three Fy(a−) and four s− were screened out in 1258 random blood samples. The multiple polymerase chain reaction‐based MS detection system can be used in rare‐blood‐type screening with good accuracy and stability.

High-Throughput Screening for Oligonucleotide Detection by ADE-OPI-MS

“…Matrix assisted laser desorption/ionization (MALDI) mass spectrometry is also a high-throughput MS screening technology for a macromolecule analysis. 13,18,19 However, MALDI-MS relies on indispensable matrices to assist the ionization of analytes. Due to the heterogeneity in the morphology of sample-matrix cocrystals, MALDI-MS has suffered from a high degree of signal variation, especially for the analysis of DNA.…”

“…Owing to these advantages, LC–MS has now become the predominant analytics of oligonucleotide identification in a low-throughput manner. − However, the extended separation time of LC–MS is not conducive to the processing of thousands of high-throughput samples. Matrix assisted laser desorption/ionization (MALDI) mass spectrometry is also a high-throughput MS screening technology for a macromolecule analysis. ,, However, MALDI-MS relies on indispensable matrices to assist the ionization of analytes. Due to the heterogeneity in the morphology of sample-matrix cocrystals, MALDI-MS has suffered from a high degree of signal variation, especially for the analysis of DNA.…”