Volume 21, Issue 2 (Summer 2024)
Sci J Iran Blood Transfus Organ 2024, 21(2): 138-150 |
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Masoudi M, Sharifi Z, Maghsudlu M. Molecular detection of HTLV-1 provirus from DNA released from cells in plasma, in cases of lack of access to genomic DNA of peripheral blood mononuclear cells. Sci J Iran Blood Transfus Organ 2024; 21 (2) :138-150
URL: http://bloodjournal.ir/article-1-1529-en.html
URL: http://bloodjournal.ir/article-1-1529-en.html
Abstract: (841 Views)
Abstract
Background and Objectives
Human lymphotropic virus type I mainly infects lymphocytes. For the detection of this virus, mainly nucleated cells are evaluated. Due to the short longevity of cells, sample collection is a challenge. The purpose of the study was to investigate the presence of HTLV-1 virus in genomic DNA released from cells in plasma and to determine its viral load in Western blot-positive blood donors.
Materials and Methods
In this descriptive study, 30 HTLV-1 positive Western blot donors were evaluated. Using an extraction kit, genomic DNA was extracted from plasma and isolated PBMC using Ficol. A nested PCR test was used to confirm the presence of HTLV-1 virus. A real-time PCR quantitative test by Sybergreen method was used to determine the amount of virus load.
Results
The average age of subjects was 42.9 ± 13.5 years; 93.3% of them were male and 6.6% female. LTR gene region was detected by nested PCR method in 80% of plasma and 100% of PBMC samples. The median viral loads in plasma and PBMC were 1.90 Copies/µL and 20 copies/106 PBMC, respectively.
Conclusions
To detect the HTLV-1 virus by molecular method, the HTLV-1 provirus DNA released from cells, in the plasma can be used when peripheral blood mononuclear cells are unavailable. It has also been observed that the virus load in the plasma of asymptomatic carriers is approximately 10 times lower than in PBMC.
Background and Objectives
Human lymphotropic virus type I mainly infects lymphocytes. For the detection of this virus, mainly nucleated cells are evaluated. Due to the short longevity of cells, sample collection is a challenge. The purpose of the study was to investigate the presence of HTLV-1 virus in genomic DNA released from cells in plasma and to determine its viral load in Western blot-positive blood donors.
Materials and Methods
In this descriptive study, 30 HTLV-1 positive Western blot donors were evaluated. Using an extraction kit, genomic DNA was extracted from plasma and isolated PBMC using Ficol. A nested PCR test was used to confirm the presence of HTLV-1 virus. A real-time PCR quantitative test by Sybergreen method was used to determine the amount of virus load.
Results
The average age of subjects was 42.9 ± 13.5 years; 93.3% of them were male and 6.6% female. LTR gene region was detected by nested PCR method in 80% of plasma and 100% of PBMC samples. The median viral loads in plasma and PBMC were 1.90 Copies/µL and 20 copies/106 PBMC, respectively.
Conclusions
To detect the HTLV-1 virus by molecular method, the HTLV-1 provirus DNA released from cells, in the plasma can be used when peripheral blood mononuclear cells are unavailable. It has also been observed that the virus load in the plasma of asymptomatic carriers is approximately 10 times lower than in PBMC.
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