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Pathogen reduction in human plasma using an ultrashort pulsed laser.

Tsen SW, Kingsley DH, Kibler K, Jacobs B, Sizemore S, Vaiana SM, Anderson J, Tsen KT, Achilefu S - PLoS ONE (2014)

Bottom Line: Pathogen reduction is a viable approach to ensure the continued safety of the blood supply against emerging pathogens.We found that laser treatment resulted in 2-log, 1-log, and 3-log reductions in human immunodeficiency virus, hepatitis A virus, and murine cytomegalovirus in human plasma, respectively.Ultrashort pulsed lasers are a promising new method for chemical-free, broad-spectrum pathogen reduction in human plasma.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiology, Washington University School of Medicine, St Louis, Missouri, United States of America.

ABSTRACT
Pathogen reduction is a viable approach to ensure the continued safety of the blood supply against emerging pathogens. However, the currently licensed pathogen reduction techniques are ineffective against non-enveloped viruses such as hepatitis A virus, and they introduce chemicals with concerns of side effects which prevent their widespread use. In this report, we demonstrate the inactivation of both enveloped and non-enveloped viruses in human plasma using a novel chemical-free method, a visible ultrashort pulsed laser. We found that laser treatment resulted in 2-log, 1-log, and 3-log reductions in human immunodeficiency virus, hepatitis A virus, and murine cytomegalovirus in human plasma, respectively. Laser-treated plasma showed ≥70% retention for most coagulation factors tested. Furthermore, laser treatment did not alter the structure of a model coagulation factor, fibrinogen. Ultrashort pulsed lasers are a promising new method for chemical-free, broad-spectrum pathogen reduction in human plasma.

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Related in: MedlinePlus

Inactivation of viruses in plasma using a USP laser.Human plasma containing HIV (A), HAV (B), or MCMV (C) were treated with the USP laser. For the HIV-spiked plasma, viral titer was assessed by plaque assay in MAGI cells. For the HAV-spiked plasma, viral titer was assessed by plaque assay in fetal rhesus monkey kidney cells. For the MCMV-spiked plasma, viral titer was assessed by TCID50 assay in murine embryonic fibroblast cells. Results are representative of triplicate experiments and are shown as means ± SEM.
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pone-0111673-g001: Inactivation of viruses in plasma using a USP laser.Human plasma containing HIV (A), HAV (B), or MCMV (C) were treated with the USP laser. For the HIV-spiked plasma, viral titer was assessed by plaque assay in MAGI cells. For the HAV-spiked plasma, viral titer was assessed by plaque assay in fetal rhesus monkey kidney cells. For the MCMV-spiked plasma, viral titer was assessed by TCID50 assay in murine embryonic fibroblast cells. Results are representative of triplicate experiments and are shown as means ± SEM.

Mentions: For this study, we chose HIV and HAV as medically significant enveloped and non-enveloped RNA viruses, respectively, and we chose MCMV as a representative enveloped DNA virus whose results could be extrapolated to relevant human pathogens such as cytomegalovirus and hepatitis B virus. To demonstrate that the USP laser treatment can inactivate viruses in plasma, aliquots of HIV, HAV, or MCMV were spiked into human plasma and treated with the laser. USP laser treatment of virus-spiked plasma samples resulted in approximately 2-log, 1-log, and 3-log reductions in HIV, HAV, and MCMV titers, respectively (Figure 1A–C). The reduction in HAV titers after USP laser treatment exceeds that achieved by the currently licensed SD and amotosalen techniques [6]. It is anticipated that further optimization of laser parameters such as wavelength (i.e., operating at wavelengths where absorption by bilirubin/hemoglobin are minimal) would yield greater inactivation of viruses. These data indicate that USP laser treatment can achieve clinically meaningful reduction of viruses in human plasma.


Pathogen reduction in human plasma using an ultrashort pulsed laser.

Tsen SW, Kingsley DH, Kibler K, Jacobs B, Sizemore S, Vaiana SM, Anderson J, Tsen KT, Achilefu S - PLoS ONE (2014)

Inactivation of viruses in plasma using a USP laser.Human plasma containing HIV (A), HAV (B), or MCMV (C) were treated with the USP laser. For the HIV-spiked plasma, viral titer was assessed by plaque assay in MAGI cells. For the HAV-spiked plasma, viral titer was assessed by plaque assay in fetal rhesus monkey kidney cells. For the MCMV-spiked plasma, viral titer was assessed by TCID50 assay in murine embryonic fibroblast cells. Results are representative of triplicate experiments and are shown as means ± SEM.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4221090&req=5

pone-0111673-g001: Inactivation of viruses in plasma using a USP laser.Human plasma containing HIV (A), HAV (B), or MCMV (C) were treated with the USP laser. For the HIV-spiked plasma, viral titer was assessed by plaque assay in MAGI cells. For the HAV-spiked plasma, viral titer was assessed by plaque assay in fetal rhesus monkey kidney cells. For the MCMV-spiked plasma, viral titer was assessed by TCID50 assay in murine embryonic fibroblast cells. Results are representative of triplicate experiments and are shown as means ± SEM.
Mentions: For this study, we chose HIV and HAV as medically significant enveloped and non-enveloped RNA viruses, respectively, and we chose MCMV as a representative enveloped DNA virus whose results could be extrapolated to relevant human pathogens such as cytomegalovirus and hepatitis B virus. To demonstrate that the USP laser treatment can inactivate viruses in plasma, aliquots of HIV, HAV, or MCMV were spiked into human plasma and treated with the laser. USP laser treatment of virus-spiked plasma samples resulted in approximately 2-log, 1-log, and 3-log reductions in HIV, HAV, and MCMV titers, respectively (Figure 1A–C). The reduction in HAV titers after USP laser treatment exceeds that achieved by the currently licensed SD and amotosalen techniques [6]. It is anticipated that further optimization of laser parameters such as wavelength (i.e., operating at wavelengths where absorption by bilirubin/hemoglobin are minimal) would yield greater inactivation of viruses. These data indicate that USP laser treatment can achieve clinically meaningful reduction of viruses in human plasma.

Bottom Line: Pathogen reduction is a viable approach to ensure the continued safety of the blood supply against emerging pathogens.We found that laser treatment resulted in 2-log, 1-log, and 3-log reductions in human immunodeficiency virus, hepatitis A virus, and murine cytomegalovirus in human plasma, respectively.Ultrashort pulsed lasers are a promising new method for chemical-free, broad-spectrum pathogen reduction in human plasma.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiology, Washington University School of Medicine, St Louis, Missouri, United States of America.

ABSTRACT
Pathogen reduction is a viable approach to ensure the continued safety of the blood supply against emerging pathogens. However, the currently licensed pathogen reduction techniques are ineffective against non-enveloped viruses such as hepatitis A virus, and they introduce chemicals with concerns of side effects which prevent their widespread use. In this report, we demonstrate the inactivation of both enveloped and non-enveloped viruses in human plasma using a novel chemical-free method, a visible ultrashort pulsed laser. We found that laser treatment resulted in 2-log, 1-log, and 3-log reductions in human immunodeficiency virus, hepatitis A virus, and murine cytomegalovirus in human plasma, respectively. Laser-treated plasma showed ≥70% retention for most coagulation factors tested. Furthermore, laser treatment did not alter the structure of a model coagulation factor, fibrinogen. Ultrashort pulsed lasers are a promising new method for chemical-free, broad-spectrum pathogen reduction in human plasma.

Show MeSH
Related in: MedlinePlus