Rapid and Reliable One-Step ABO Genotyping Using Direct Real-Time Allele-Specific PCR and Melting Curve Analysis Without DNA Preparation

Indian Journal of Hematology & Blood Transfusion : an Official Journal of Indian Society of Hematology and Blood Transfusion
Jun-Hee ParkGeon Park

Abstract

ABO genotyping is a molecular diagnostic technique important for transfusion and transplantation in medicine, and human identification in forensic science. Because ABO genotyping are labor intensive and time consuming, the genotyping cannot be firstly used to resolve the serological ABO discrepancy in blood bank. For rapid one-step ABO genotyping, we developed direct, real-time, allele-specific polymerase chain reaction (PCR), and melting curve analysis (DRAM assay) without DNA preparation. In DRAM assay, we used a special PCR buffer for direct PCR, a rapid RBC lysis buffer, white blood cells as template without DNA preparation, allele-specific primers for discriminating three ABO alleles (261G/del, 796C/A, and 803G/C), and melting curve analysis as a detection method. There was 100% concordance among the results of ABO genotyping by the DRAM assay, serologic typing, PCR-RFLP and PCR-direct sequencing of 96 venous blood samples. We were able to reduce the number of manual steps to three and the hands-on time to 12 min, compared to seven steps and approximately 40 min for conventional ABO genotyping using allele-specific PCR with purified DNA and agarose gel electrophoresis. We have established and validated the DRAM assay for r...Continue Reading

References

Aug 1, 1992·Human Molecular Genetics·P H Johnson, D A Hopkinson
Jun 28, 2003·Blood·Axel SeltsamRainer Blasczyk
May 26, 2004·Transfusion·Gianmarco FerriSusi Pelotti
Mar 3, 2007·Clinica Chimica Acta; International Journal of Clinical Chemistry·Young Geun YangDoo-Sik Kim
Jul 3, 2009·The Korean journal of laboratory medicine·Sung Ho LeeSuhng Wook Kim
Dec 17, 2009·Journal of Forensic Sciences·Li RuanQingge Li
Jan 16, 2010·The Journal of Molecular Diagnostics : JMD·Zhian ZhangWayne M Barnes
Jan 14, 2011·Journal of Forensic Sciences·Hwan Young LeeKyoung-Jin Shin
Jan 14, 2011·Journal of Forensic Sciences·Ken WatanabeKoichi Sakurada
Jul 16, 2011·Clinica Chimica Acta; International Journal of Clinical Chemistry·Seon Young KimJunghan Song
Dec 20, 2011·Legal Medicine·Tomonori MuroHaruo Takeshita
Jun 12, 2012·Molecular and Cellular Probes·Hirofumi TsutsumiToshinobu Komuro
Oct 17, 2012·Legal Medicine·Kazuho MaedaKatsuyoshi Kurihara
Oct 24, 2017·Forensic Science International. Genetics·Sarah E Cavanaugh, Abigail S Bathrick

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