Anti-primer quenching-based real-time PCR for simplex or multiplex DNA quantification and single-nucleotide polymorphism genotyping

2007 ◽  
Vol 2 (1) ◽  
pp. 50-58 ◽  
Author(s):  
Jin Li ◽  
G Mike Makrigiorgos
Keyword(s):  
Single Nucleotide Polymorphism ◽  
Real Time ◽  
Real Time Pcr ◽  
Dna Quantification ◽  
Single Nucleotide Polymorphism Genotyping ◽  
Nucleotide Polymorphism ◽  
Single Nucleotide

scholarly journals Donor Chimerism Study by Single Nucleotide Polymorphism using SYBR green based Real Time PCR

2021 ◽  
Vol 37 (7) ◽  
Author(s):  
Ayesha Nayyar ◽  
Suhaib Ahmed
Keyword(s):  
Single Nucleotide Polymorphism ◽  
Real Time ◽  
Real Time Pcr ◽  
Sybr Green ◽  
Haematopoietic Stem Cell ◽  
Nucleotide Polymorphism ◽  
Green Method ◽  
Single Nucleotide ◽  
Donor Chimerism ◽  
Haematological Disorders

Objective: To optimize and evaluate a real time PCR of Single Nucleotide Polymorphism by SYBR Green method for detection of donor chimerism after haematopoietic stem cell transplantation. Methods: This descriptive study was conducted at Genetic Resource Centre (GRC) Lab Rawalpindi from Oct 2017 - Dec 2019. A total of twenty patients of post haematopoietic stem cell transplant with various haematological disorders were studied to see the status of donor chimerism by using SNP real time PCR using SYBR Green method and short tandem repeat PCR. These patients had undergone allogeneic HSCT from HLA-matched sibling donors at Pakistan Institute of Medical Science and Armed Forces Bone Marrow Transplant Centre. Results: Real time PCR using SYBR Green was able to detect significant amount of chimerism in all 20 patients having undergone HSCT. Regarding precision of the real time PCR assay the mean value of donor chimerism was 94.1% (SD 3.96) and by STR PCR it was 95.1% (SD 1.41). The assay was found to be sensitive with a detection limit of <1%. Conclusion: Our results demonstrate that SNP analysis by SYBR Green real time PCR may be used for the evaluation of chimerism status in patients having undergone HSCT with a sensitivity of <1%. Hence donor chimerism by this sensitive method can be used in monitoring of chimerism in post-transplant patients with various haematological disorders. doi: https://doi.org/10.12669/pjms.37.7.4203 How to cite this:Nayyar A, Ahmed S. Donor Chimerism Study by Single Nucleotide Polymorphism using SYBR green based Real Time PCR. Pak J Med Sci. 2021;37(7):---------. doi: https://doi.org/10.12669/pjms.37.7.4203 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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