scholarly journals Single Nucleotide Polymorphism (SNP) Genotyping in Unprocessed Whole Blood and Serum by Real-Time PCR

2001 ◽  
Vol 47 (11) ◽  
pp. 2050-2053 ◽  
Author(s):  
Arve Ulvik ◽  
Per Magne Ueland
2020 ◽  
Vol 14 (01) ◽  
pp. 80-88
Author(s):  
Amal E Saafan ◽  
Ashraf Abobaker ◽  
Mohamed S Abbas ◽  
Ahmed El-Gendy

Introduction: In Egypt, 15% of the populations are suffering from chronic hepatitis C especially genotype 4. Sofosbuvir was approved by FDA in December 2013 for treatment of HCV genotypes 2 and 3 in combination with Ribavirin, and for genotypes 1 and 4 in combination with Peg-IFN. Recently, polymorphism of different genes and plasma levels of IL-6 were utilized for better prediction of HCV clearance. This study aimed at early prediction of the efficacy of HCV treatment with Sofosbuvir (Sovaldi) and comparing the antiviral efficacy of dual and triple Sovaldi combination therapy. Methodology: Blood samples were collected from 100 HCV positive patients and detected by real time PCR at three time intervals. SNP genotyping of INFL-4 gene was estimated by using real-time PCR with predesigned primers and Taqman probes. IL-6 serum level was estimated before, during and after the end of the treatment using ELISA assay based on human IL-6 KIT. Results: SNP genotyping of INFL-4 gene showed that 13.1% of patients carried ∆G/∆G, 30.4% patients had TT/TT and 56.5% patients possessed heterozygote allele ∆G/TT. Clinical data displayed that 13 patients were got relapsed at SVR 12. Serum level of IL-6 was noticed higher in HCV patients than healthy ones. Noteworthy, it was increased during treatment then decreased to a minimal level than begining of treatment. Conclusion: SNP in INFL-4 gene has displayed no effect in response to Sofosbuvir. Dual therapy had the same effect like triple therapy, so interferon could be withdrawn from the treatment regimen.


2021 ◽  
Vol 37 (7) ◽  
Author(s):  
Ayesha Nayyar ◽  
Suhaib Ahmed

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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