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Determination of ABO blood group genotypes using the real‑time loop‑mediated isothermal amplification method.

Zhang C, Zhu J, Yang J, Wan Y, Ma T, Cui Y - Mol Med Rep (2015)

Bottom Line: ABO genotyping is commonly used in several situations, including blood transfusion, personal identification and disease detection.The results were compared with the phenotypes determined by serological assay and the genotypes determined by direct sequencing, and no discrepancies were observed.This novel and rapid method, with good accuracy and reasonably cost effective, provides a supplement to routine serological ABO typing and may also be useful in other point‑of‑care testing.

View Article: PubMed Central - PubMed

Affiliation: College of Life Sciences, Northwest University, Xi'an, Shaanxi 710069, P.R. China.

ABSTRACT
ABO genotyping is commonly used in several situations, including blood transfusion, personal identification and disease detection. The present study developed a novel method for ABO genotyping, using loop‑mediated isothermal amplification (LAMP). This method allows the simultaneous determination of six ABO genotypes under 40 min at a constant temperature of 62˚C. The genotypes of 101 blood samples were determined to be AA (n=6), AO (n=38), BB (n=12), BO (n=29), AB (n=8) and OO (n=8) by the LAMP assay. The results were compared with the phenotypes determined by serological assay and the genotypes determined by direct sequencing, and no discrepancies were observed. This novel and rapid method, with good accuracy and reasonably cost effective, provides a supplement to routine serological ABO typing and may also be useful in other point‑of‑care testing.

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Schematic of primers and the process for real-time loop-mediated isothermal amplification using the O primer set used as an example.
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f1-mmr-12-04-5963: Schematic of primers and the process for real-time loop-mediated isothermal amplification using the O primer set used as an example.

Mentions: Two single nucleotide polymorphisms (SNPs) at nucleotides 261 and 803, located on exons 6 and 7 of the ABO gene, which cover the most polymorphic sites of the complete ABO sequences, as shown in Fig. 1 (9), were selected. The primer set for LAMP amplification includes a set of four primers, comprising two outer and two internal primers, which recognize six distinct regions on the target sequence, designated in Fig. 1. Table I lists the sequences of the typing primers used for the LAMP reaction. All oligonucleotide primers were synthesized by Beijing Genomics Institute (BGI; Beijing, China).


Determination of ABO blood group genotypes using the real‑time loop‑mediated isothermal amplification method.

Zhang C, Zhu J, Yang J, Wan Y, Ma T, Cui Y - Mol Med Rep (2015)

Schematic of primers and the process for real-time loop-mediated isothermal amplification using the O primer set used as an example.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581819&req=5

f1-mmr-12-04-5963: Schematic of primers and the process for real-time loop-mediated isothermal amplification using the O primer set used as an example.
Mentions: Two single nucleotide polymorphisms (SNPs) at nucleotides 261 and 803, located on exons 6 and 7 of the ABO gene, which cover the most polymorphic sites of the complete ABO sequences, as shown in Fig. 1 (9), were selected. The primer set for LAMP amplification includes a set of four primers, comprising two outer and two internal primers, which recognize six distinct regions on the target sequence, designated in Fig. 1. Table I lists the sequences of the typing primers used for the LAMP reaction. All oligonucleotide primers were synthesized by Beijing Genomics Institute (BGI; Beijing, China).

Bottom Line: ABO genotyping is commonly used in several situations, including blood transfusion, personal identification and disease detection.The results were compared with the phenotypes determined by serological assay and the genotypes determined by direct sequencing, and no discrepancies were observed.This novel and rapid method, with good accuracy and reasonably cost effective, provides a supplement to routine serological ABO typing and may also be useful in other point‑of‑care testing.

View Article: PubMed Central - PubMed

Affiliation: College of Life Sciences, Northwest University, Xi'an, Shaanxi 710069, P.R. China.

ABSTRACT
ABO genotyping is commonly used in several situations, including blood transfusion, personal identification and disease detection. The present study developed a novel method for ABO genotyping, using loop‑mediated isothermal amplification (LAMP). This method allows the simultaneous determination of six ABO genotypes under 40 min at a constant temperature of 62˚C. The genotypes of 101 blood samples were determined to be AA (n=6), AO (n=38), BB (n=12), BO (n=29), AB (n=8) and OO (n=8) by the LAMP assay. The results were compared with the phenotypes determined by serological assay and the genotypes determined by direct sequencing, and no discrepancies were observed. This novel and rapid method, with good accuracy and reasonably cost effective, provides a supplement to routine serological ABO typing and may also be useful in other point‑of‑care testing.

Show MeSH