{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T23:14:55Z","timestamp":1783466095565,"version":"3.55.0"},"reference-count":26,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,5,12]],"date-time":"2022-05-12T00:00:00Z","timestamp":1652313600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81671476 31570906"],"award-info":[{"award-number":["81671476 31570906"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Genet."],"abstract":"<jats:p><jats:bold>Background:<\/jats:bold> Human neutrophil antigen-3A (HNA-3A) and human neutrophil antigen-3B (HNA-3B) are generated by a single-nucleotide polymorphism (rs2288904, c.461G &amp;gt; A) in exon 7 of the choline transporter-like protein-2 gene (<jats:italic>CTL2<\/jats:italic>, also known as <jats:italic>SLC44A2<\/jats:italic>). Antibodies to HNA-3 can be generated following blood transfusion or other factors resulting in exposure to HNA-3 antigens. These antibodies can cause transfusion-related acute lung injury (TRALI) or neonatal alloimmune neutropenia (NAIN). This study describes a sensitive and specific TaqMan real-time polymerase chain reaction (PCR) method to screen for the HNA-3 genotype using specific primers and probes designed to detect allelic polymorphisms. Considering the high sensitivity and accuracy of droplet digital PCR (ddPCR) in the identification of the rare <jats:italic>SLC44A2*2<\/jats:italic> allele, we used this technique to identify blood donors with the rare HNA-3B antigen and calculate the allele frequency of <jats:italic>SLC44A2<\/jats:italic> in mixed populations with different proportions.<\/jats:p><jats:p><jats:bold>Methods:<\/jats:bold> DNA samples purified from 208 donors in northwest China were subjected to TaqMan real-time PCR to detect allelic polymorphisms in <jats:italic>SLC44A2<\/jats:italic>. The results were confirmed by Sanger sequencing. The rare HNA-3B antigen was detected by ddPCR. <jats:italic>SLC44A2<\/jats:italic> frequency was determined by two-channel ddPCR.<\/jats:p><jats:p><jats:bold>Results:<\/jats:bold> The genotypes of all DNA samples were detected by the TaqMan real-time PCR using specific probes for HNA-3, and the results were consistent with the Sanger sequencing results in respect to the HNA-3A and HNA-3B polymorphisms. The allele frequencies of <jats:italic>SLC44A2*1<\/jats:italic> and <jats:italic>SLC44A2*2<\/jats:italic> in the 208 donors in northwest China were 64.9% (95% confidence interval [CI], 59%\u201370.8%) and 35.1% (95% CI, 29.2%\u201341%), respectively. The ratio of <jats:italic>SLC44A2*2<\/jats:italic> alleles was accurately detected in all blood pools by ddPCR but not by TaqMan real-time PCR. This allowed for the <jats:italic>SLC44A2<\/jats:italic> frequency in the population to be accurately inferred.<\/jats:p><jats:p><jats:bold>Conclusion:<\/jats:bold> This new method of detecting <jats:italic>SLC44A2<\/jats:italic> alleles was highly sensitive and specific, as confirmed by Sanger sequencing. ddPCR using the designed probes resulted in successful detection of the rare HNA-3B antigen. Furthermore, we successfully detected the rare HNA-3B antigen and inferred the <jats:italic>SLC44A2<\/jats:italic> frequency by ddPCR using the probes that we designed.<\/jats:p>","DOI":"10.3389\/fgene.2022.794285","type":"journal-article","created":{"date-parts":[[2022,5,12]],"date-time":"2022-05-12T09:17:05Z","timestamp":1652347025000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":3,"title":["SLC44A2 Frequency, a New TaqMan Real-Time Polymerase Chain Reaction Method for HNA-3A\/3B Genotyping, and a New Application of Droplet Digital PCR"],"prefix":"10.3389","volume":"13","author":[{"given":"Yufeng","family":"Wang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xihui","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qi","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tangdong","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kun","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanming","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2022,5,12]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"778","DOI":"10.1111\/bjh.17295","article-title":"Serologic and Molecular Studies to Identify Neonatal Alloimmune Neutropenia in a Cohort of 10,000 Neonates","volume":"192","author":"Abbas","year":"2021","journal-title":"Br. 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