Original Article

Inactivation of model viruses and bacteria in human fresh frozen plasma using riboflavin and long wave ultraviolet rays


Background and Objectives: Pathogen reduction technologies are among methods to eliminate transfusion transmitted infections. Mirasol method using riboflavin in combination with ultraviolet rays is one of them. The aims of this study were to investigate the effectiveness of Mirasol method to inactivate some model pathogens as well as examination of the sensitivity of plasma proteins after treatment.
Materials and Methods: Riboflavin in 50μM concentration and ultraviolet (365 nm) in three different energy doses (3.6, 7.2, and 10.8 j/cm2) were employed to inactivate model pathogens. Four standard viruses were used in this study including Vesicular Stomatitis Virus (VSV), Herpes Simplex Virus1 (HSV-1), Bovine Viral Diarrhea Virus (BVDV) and Polio Virus. 50% Tissue Culture Infectious Dose (TCID50) and Reed–Muench Methods were used to estimate viruses’ titers. E. coli and Staphylococcus aureus were used as bacterial models. Four plasma proteins including factor V, VIII, fibrinogen and antithromin were used to determine their sensitivity to pathogen inactivation treatment.
Results: The most pathogen reduction titre was determined for 15 minutes irradiation period equal to 10.8 J/cm2 that is corresponding to Log 6.10 for BVDV, Log 6.09 for HSV-1, Log 6.62 for VSV and Log 3.36 for Polio. Bacterial reduction titer was Log 6.94 for E. coli and Log 7.00 for S. aureus. Indicator proteins for plasma activity were determined to be 75% for factor V, 88% for factor VIII, 52% for fibrinogen and 94% for antithrombin.
Conclusion: Results showed that the employed method can inactivate most of the pathogens in fresh frozen plasma. The acceptable activities of selected plasma proteins remained after treatment.


Busch M, Kleinman S, Nemo G. Current and emerging infectious risks of blood Transfusion. J Am Med Assoc 2003; 289:959-62.

Goodrich R, Edrich R, Li J, Seghatchian J. The Mirasol PRT system for pathogen reduction of platelets and plasma: an overview of current status and future trends. Transfusion and Apheresisi Science 2006; 35:5-17.

Vanlandingham D, Keil Sh, McElroy K, Pyles R, Goodrich R, Higgs S. Photochemical inactivation of chikungunya virus in plasma and platelets using the Mirasol pathogen reduction technology system. Transfusion 2013; 88:1163-69.

Wainwright M, Baptista M. The application of photosensitisers to tropical pathogens in the blood supply. Photodiagnosis and Photodynamic Therapy 2011; 8: 240-248.

Marschner S, Goodrich R. Pathogen reduction technology treatment of platelets, plasma and whole blood using riboflavin and UV light. Transfusion Medicine and Hemotherapy 2011; 38:8-18.

Hayashi M. Kishi M, Sofuni T, Ishidate M Jr. Micronucleus tests in mice on 39 food additive and eight miscellaneous chemicals. Food Chem Toxicol 1988; 26:487-500.

Munoz N, Hayashi M, Bang LJ, Wahrendorf J, Crespi M, Bosch FX. Effect of riboflavin, retinol and Zinc on micronuclei of buccal mucosa and of esophagus: a randomized double – blind intervention study in China. J Natl Cancer Inst 1987; 79:687-691.

Unna K, Greslin JG. Studies on the toxicity and pharmacology of riboflavin. J Pharmacol 1942; 76:75-80.

Li J, De Korte D, Woolum MD, et al. Pathogen reduction of buffy coat platelet concentrates using riboflavin and light: comparisons with pathogen-reduction technology- treated apheresis platelet products. Vox Sang 2004; 87:82-90.

Ruane PH, Edrich R, Gampp D, Keil SD, Leonard RL, Goodrich RP. Photochemical inactivating of selected viruses and bacteria in platelet concentrates using riboflavin and light. Transfusion 2004; 44: 877-85.

Goodrich RP,Gilmour D, Hovenga N, Keil SD. A laboratory comparison of pathogen reduction technology treatment and concerns. Transfusion 2009; 49:1205-16.

Aghaie A, Pourfatollah A, Bathaei S.Z, Moazzeni S.M, Khorsand Mohammad pour H, Sharifi Z. Inactivation of virus in intravenous immunoglobulin G using solvent/detergent treatment and pasteurization. Human antibodies 2008; 17: 79-84.

Reed L.J, Muench H. A simple method of estimating fifty per cent endpoints. The American journal of hygiene 1937; 27:493-497.

Goodrich R, Custer B, Keil SH, Bussch M. Defining adequate pathogen performance for transfused blood components. Transfusion 2010; 50: 1827-37.

Aubuchon J.p. Update on the status of pathogen inactivation methods. ISBT Science Series 2011; 6:181-188.

Goodrich R, Doane S, Heather R. Design and development of a method for the reduction of infectious pathogen load an inactivation of white blood cells in whole blood products. Biologicals 2009; 1-11.

Antic A, Stanojkovic Z, Macukanovic-Golubovic L, Jelic M. Evaluation of coagulation factors in fresh frozen plasma treated with riboflavin and ultra violet light. Vojnosanit Pregl 2012; 69: 22-26.

IssueVol 9 No 1 (2017) QRcode
SectionOriginal Article(s)
Viruses Bacteria Fresh Frozen Plasma Riboflavin Ultraviolet rays Inactivation

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How to Cite
Elikaei A, Hosseini SM, Sharifi Z. Inactivation of model viruses and bacteria in human fresh frozen plasma using riboflavin and long wave ultraviolet rays. Iran J Microbiol. 2017;9(1):50-54.