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  5. Reducing Threats to the Blood Supply from West Nile Virus, Dengue Virus, and Chikungunya Virus Through Development of Detection Tools and Studies of Genetic Evolution and Pathogenesis
  1. Biologics Research Projects

Reducing Threats to the Blood Supply from West Nile Virus, Dengue Virus, and Chikungunya Virus Through Development of Detection Tools and Studies of Genetic Evolution and Pathogenesis

Principal Investigator: Maria Rios, PhD
Office / Division / Lab: OBRR / DETTD / LEP

General Overview

The mission of our research program is to reduce the risk of transmission of West Nile Virus (WNV), dengue virus (DENV), chikungunya virus (CHIKV), and the newly recognized threat Zika virus (ZIKV), through donated blood. This work is important because these viruses together have a substantial public health impact. ZIKV has caused large outbreaks in the Pacific region and the Americas since 2013, causes the Guillain-Barre syndrome and, when infecting pregnant women, birth defects such as microcephaly. Outbreaks of WNV in the U.S. have caused illness in tens of thousands of people, including ~20,000 cases of neurological disease, and >1,800 deaths through 2015. DENV causes 50-100 million cases of disease and tens of thousands of deaths annually worldwide. The first CHIKV outbreak in the Americas began in late 2013 in the Caribbean and quickly spread to South, Central and North American countries, causing >2 million cases by mid-2016. No effective therapies or FDA-licensed vaccines exist for these human infections. ZIKV, CHIKV and DENV are not yet established in the U.S., although small outbreaks have been caused by DENV in Hawaii, Texas and New York, both DENV and CHIKV in Florida, and all three have affected U.S. territories. Infected travelers and inadvertent importation of infected mosquitoes pose a risk, since the transmitting vectors (Aedes mosquitoes) are present in the contiguous U.S.

These viruses frequently cause asymptomatic (silent) infections, during which affected individuals feel well enough to donate blood. Transfusion-transmitted (TT) WNV, DENV and ZIKV have been reported, and TT-CHIKV is suspected to have occurred. Symptomatic infections may include headache, rash, conjunctivitis, joint and muscle pain and fever, and are often mistaken for the common cold; however, they may evolve into serious, fatal diseases.

To address the potential for WNV transmission through transplants and transfusions, FDA collaborated with industry and blood banks to develop new donor screening tests, which were in use by summer 2003 under an FDA-approved research protocol.

In response to the threat of these viruses, our laboratory is 1) preparing standardized virus stocks for evaluation of the accuracy of commercially-developed tests designed to identify these viruses in blood prior to their release for marketing and quality control by FDA; 2) studying the genetic variability of these viruses in order to identify mutants that could escape detection by these tests or interfere with vaccine development; 3) developing and evaluating technologies that enhance the ability of tests to identify the genetic variability of these pathogens in small volumes of specimen; 4) studying how the viruses interact with cellular components of blood, including red blood cells (RBCs) and 5) studying how the viruses cause disease, which might produce new information that could help in the search for new approaches for pathogen inactivation and therapies.

Our regulatory research also includes the development of renewable DNA reference panels to evaluate the accuracy of commercially developed blood donor molecular genotyping kits. These kits are used to enhance matching of RBC units for patients who need frequent transfusions, such as Sickle Cell Disease patients.

Scientific Overview

Our research focuses on understanding human infection with arthropod-borne viruses (arboviruses) relevant to transfusion safety, such as Zika virus (ZIKV), chikungunya virus (CHIKV), Dengue virus (DENV), and West Nile virus (WNV). These viruses present a challenge to the scientific community because these diseases can be fatal but there are no specific therapies or FDA-licensed vaccines available. These viruses are commonly transmitted by mosquitoes; ZIKV, CHIKV and DENV by Aedes sp., and WNV by Culex sp. ZIKV can also be sexually transmitted, and transfusion-transmission has been reported for all but CHIKV, which can be potentially transmitted by that route. Most infections with ZIKV, DENV and WNV, and many with CHIKV, are asymptomatic, despite the presence of viremia.

WNV is endemic in the contiguous U.S. where the blood supply is screened by WNV-NAT. Additionally, local transmissions of CHIKV in Florida, and DENV in Florida, South Texas and New York have occurred. A DENV outbreak occurred in Hawaii in 2015-16, and DENV is endemic to Puerto Rico and other U.S. territories. No locally-transmitted mosquito-borne cases of ZIKV have been reported in the U.S. There is also evidence that arbovirus mutations (CHIKV in Italy and WNV in the U.S.), enabled new variants to spread more efficiently.

Travel and U.S. troop deployment to areas where arboviruses are endemic, have raised concerns that these diseases could be inadvertently imported and spread by mosquitoes already established in the U.S. This threat has prompted development of new blood-screening assays designed to keep the U.S. blood supply safe for transfusions. Reference reagents for those agents assist development and validation of new assays; aid the determination of sensitivity and specificity to meet FDA regulatory requirements for licensure, and ensure consistency of new lots prior to market. A major roadblock is the lack of large volumes of well-characterized viral stocks, which cannot be obtained by direct human sampling.

This research program aims to: 1) produce reference panels needed to promote transfusion safety and update existing standard reagents as needed for FDA testing proficiency and lot-release panels; 2) identify the potential impact of genetic mutations of viruses on the performance of diagnostic and screening assays, and the efficacy of vaccines and therapies under development; 3) investigate the relevance of genetic variation to virus fitness and virulence using in vitro culture systems with human and animal cells; 4) assess potential ways to increase the sensitivity of current assays to improve blood safety; 5) investigate the biological properties and significance of viral association with blood cells (primarily RBCs) and their significance for infection by transfusion; 6) develop technology to enhance detection of arboviruses and predict infection outcomes.

These studies will cover unmet needs such as reference materials to assist the development and licensure of assays, and data to assess the efficacy of blood screening by estimation of residual risks after their implementation, and support decision-making on issues for which clinical evidence is not available, including blood safety, transplantation and vaccines.


Transfusion 2017 Aug;57(8):1977-87
Collaborative study to establish World Health Organization international reference reagents for dengue virus types 1 to 4 RNA for use in nucleic acid testing.
Anez G, Volkova E, Jiang Z, Heisey DAR, Chancey C, Fares RCG, Rios M, Collaborative Study Group

J Mol Diagn 2017 Jul;19(4):549-60
Highly multiplex real-time PCR-based screening for blood-borne pathogens on an OpenArray platform.
Grigorenko E, Fisher C, Patel S, Winkelman V, Williamson P, Chancey C, Anez G, Rios M, Majam V, Kumar S, Duncan R

Antiviral Res 2017 May;141:19-28
Strategies for serum chemokine/cytokine assessment as biomarkers of therapeutic response in HCV patients as a prototype to monitor immunotherapy of infectious diseases.
Menezes EG, Coelho-Dos-Reis JG, Cardoso LM, Lopes-Ribeiro A, Jonathan-Goncalves J, Porto Goncalves MT, Cambraia RD, Soares EB, Silva LD, Peruhype-Magalhaes V, Rios M, Chancey C, Teixeira-Carvalho A, Martins-Filho OA, Teixeira R

Transfusion 2017 Mar;57(3pt2):734-47
Relative analytical sensitivity of donor nucleic acid amplification technology screening and diagnostic real-time polymerase chain reaction assays for detection of Zika virus RNA.
Stone M, Lanteri MC, Bakkour S, Deng X, Galel SA, Linnen JM, Munoz-Jordán JL, Lanciotti RS, Rios M, Gallian P, Musso D, Levi JE, Sabino EC, Coffey LL, Busch MP

Transfusion 2017 Mar;57(3pt2):748-61
Harmonization of nucleic acid testing for Zika virus: development of the 1st World Health Organization International Standard.
Baylis SA, Hanschmann KO, Schnierle BS, Trosemeier JH, Blumel J, Zika Virus Collaborative Study Group

Emerg Infect Dis 2017 Jan;23(1):115-8
Meningitis associated with simultaneous infection by multiple dengue virus serotypes in children, Brazil.
Marinho PE, Bretas de Oliveira D, Candiani TM, Crispim AP, Alvarenga PP, Castro FC, Abrahão JS, Rios M, Coimbra RS, Kroon EG

J Virol Methods 2017 Jan;239:17-25
Development of a microarray-based assay for rapid detection and monitoring of genetic variants of West Nile virus circulating in the United States.
Grinev A, Chancey C, Volkova E, Chizhikov v, Rios M

PLoS Negl Trop Dis 2016 May 16;10(5):e0004717
Genetic variability of West Nile virus in U.S. blood donors from the 2012 epidemic season.
Grinev A, Chancey C, Volkova E, Anez G, Heisey DA, Winkelman V, Foster GA, Williamson P, Stramer SL, Rios M

J Appl Microbiol 2016 Apr;120(4):1119-29
Standardized methods to generate mock (spiked) clinical specimens by spiking blood or plasma with cultured pathogens.
Dong M, Fisher C, Anez G, Rios M, Nakhasi HL, Hobson JP, Beanan M, Hockman D, Grigorenko E, Duncan R

J Womens Health 2016 Mar;25(3):222-34
The Food and Drug Administration Office of Women's Health: impact of science on regulatory policy: an update.
Elahi M, Eshera N, Bambata N, Barr H, Lyn-Cook B, Beitz J, Rios M, Taylor DR, Lightfoote M, Hanafi N, DeJager L, Wiesenfeld P, Scott PE, Fadiran EO, Henderson MB

PLoS Negl Trop Dis 2016 Feb 12;10(2):e0004445
Distribution of dengue virus types 1 and 4 in blood components from infected blood donors from Puerto Rico.
Anez G, Heisey DA, Chancey C, Fares RC, Espina LM, Souza KP, Teixeira-Carvalho A, Krysztof DE, Foster GA, Stramer SL, Rios M

Hum Vaccin Immunother 2016 Feb;12(2):491-502
Booster dose after 10 years is recommended following 17DD-YF primary vaccination.
Campi-Azevedo AC, Costa-Pereira C, Antonelli LR, Fonseca CT, Teixeira-Carvalho A, Villela-Rezende G, Santos RA, Batista MA, Campos FM, Pacheco-Porto L, Junior OA, Hossell DM, Coelho-Dos-Reis JG, Peruhype-Magalhaes V, Costa-Silva MF, de Oliveira JG, Farias RH, Noronha TN, Lemos JA, von Doellinger VD, Simoes M, de Souza MM, Malaquias LC, Persi HR, Pereira JM, Martins JA, Dornelas-Ribeiro M, Vinhas AA, Alves TR, Maia ML, Freire MD, Martins RM, Homma A, Romano AP, Domingues CM, Tauil PL, Vasconcelos PF, Rios M, Caldas IR, Camacho LA, Martins-Filho OA

Genome Announc 2016 Feb 11;4(1):e01583-15
Complete genome sequences of dengue virus type 1 to 4 strains used for the development of CBER/FDA RNA reference reagents and WHO international standard candidates for nucleic acid testing.
Anez G, Heisey DA, Volkova E, Rios M

Biomed Res Int 2015;2015:321873
Epidemiological scenario of Dengue in Brazil.
Fares RC, Souza KP, Anez G, Rios M

Genome Announc 2014 Oct 30;2(5):e00811-14
Complete Genome Sequence of West Nile Virus Strains Used for the Formulation of CBER/FDA RNA Reference Reagents and Lot Release Panels for Nucleic Acid Testing.
Grinev A, Anez G, Rios M

Clin Infect Dis 2014 Jul 1;59(1):16-23
Rapid detection of hepatitis B virus in blood plasma by a specific and sensitive loop-mediated isothermal amplification assay.
Nyan DC, Ulitzky LE, Cehan N, Williamson P, Winkelmann V, Rios M, Taylor DR

Genome Announc 2014 Jul 31;2(4)
Complete Coding Region Sequence of a Chikungunya Virus Strain Used for Formulation of CBER/FDA RNA Reference Reagents for Nucleic Acid Testing.
Anez G, Heisey DA, Rios M

J Biomed Mater Res A 2014 Feb;102(2):305-14
Different molecular weight hyaluronic acid effects on human macrophage interleukin 1beta production.
Baeva LF, Lyle DB, Rios M, Langone JJ, Lightfoote MM

J Mol Diagn 2014 Jan;16(1):136-44
Multiplex screening for blood-borne viral, bacterial, and protozoan parasites using an OpenArray platform.
Grigorenko E, Fisher C, Patel S, Chancey C, Rios M, Nakhasi HL, Duncan RC

Int J Environ Res Public Health 2013 Sep 23;10(9):4486-506
Genetic analysis of West Nile virus isolates from an outbreak in Idaho, United States, 2006-2007.
Grinev A, Chancey C, Añez G, Ball C, Winkelman V, Williamson P, Foster GA, Stramer SL, Rios M

Biomed Res Int 2013;2013:678645
Dengue in the United States of America: a worsening scenario?
Anez G, Rios M

PLoS Negl Trop Dis 2013 May 30;7(5):e2245
Evolutionary dynamics of West Nile virus in the United States, 1999-2011: phylogeny, selection pressure and evolutionary time-scale analysis.
Anez G, Grinev A, Chancey C, Ball C, Akolkar N, Land KJ, Winkelman V, Stramer SL, Kramer LD, Rios M

Transfusion 2012 Sep;52(9):1949-56
Polymerase chain reaction-based tests for pan-species and species-specific detection of human Plasmodium parasites.
Mahajan B, Zheng H, Pham PT, Sedegah MY, Majam VF, Akolkar N, Rios M, Ankrah I, Madjitey P, Amoah G, Addison E, Quakyi IA, Kumar S

Am J Trop Med Hyg 2012 Sep;87(3):548-53
Phylogenetic analysis of dengue virus types 1 and 4 circulating in Puerto Rico and Key West, Florida, during 2010 epidemics.
Añez G, Heisey DA, Espina LM, Stramer SL, Rios M

J Virol Methods 2012 Aug;183(2):219-23
Application of a full-genome microarray-based assay for the study of genetic variability of West Nile virus.
Grinev A, Lu Z, Chizhikov V, Rios M

Vox Sang 2012 Aug;103(2):99-106
Distribution of hepatitis C virus in circulating blood components from blood donors.
Chancey C, Winkelman V, Foley JB, Silberstein E, Teixeira-Carvalho A, Taylor DR, Rios M

Vox Sang 2012 Jul;103(Suppl. S1):62-3
Dengue virus and other arboviruses: a global view of risks.
Rios M, Anez G, Chancey C, Grinev A

PLoS One 2011;6(11):e27459
Circulation of different lineages of dengue virus type 2 in central america, their evolutionary time-scale and selection pressure analysis.
Anez G, Morales-Betoulle ME, Rios M

BMC Immunol 2010 Jan 20;11:3
Anti-West Nile virus activity of in vitro expanded human primary natural killer cells.
Zhang M, Daniel S, Huang Y, Chancey C, Huang Q, Lei YF, Grinev A, Mostowski H, Rios M, Dayton A

ISBT Sci Ser 2009;4(1):87-94
Climate change and vector-borne viral diseases potentially transmitted by transfusion
Rios M

J Virol Methods 2008 Dec;154(1-2):27-40
Microarray-based assay for the detection of genetic variations of structural genes of West Nile virus.
Grinev A, Daniel S, Laassri M, Chumakov K, Chizhikov V, Rios M

J Infect Dis 2008 Nov 1;198(9):1300-8
In Vitro Evaluation of the Protective Role of Human Antibodies to West Nile Virus (WNV) Produced during Natural WNV Infection.
Rios M, Daniel S, Dayton AI, Wood O, Hewlett IK, Epstein JS, Caglioti S, Stramer SL

Emerg Infect Dis 2008 Mar;14(3):436-44
Genetic Variability of West Nile Virus in US Blood Donors, 2002-2005.
Grinev A, Daniel S, Stramer S, Rossmann S, Caglioti S, Rios M

Clin Infect Dis 2007 Jul 15;45(2):181-6
West nile virus adheres to human red blood cells in whole blood.
Rios M, Daniel S, Chancey C, Hewlett IK, Stramer SL

Neurology 2007 Jan 16;68(3):206-13
Differentiation of HAM/TSP from patients with multiple sclerosis infected with HTLV-I.
Puccioni-Sohler M, Yamano Y, Rios M, Carvalho SM, Vasconcelos CC, Papais-Alvarenga R, Jacobson S

Vox Sang 2006 Jul;91(1):81-7
Dombrock gene analysis in Brazilian people reveals novel alleles.
Baleotti W Jr, Rios M, Reid ME, Hashmi G, Fabron A Jr, Pellegrino J Jr, Castilho L

Transfusion 2006 Apr;46(4):659-67
Monocytes-macrophages are a potential target in human infection with West Nile virus through blood transfusion.
Rios M, Zhang MJ, Grinev A, Srinivasan K, Daniel S, Wood O, Hewlett IK, Dayton AI

J Med Virol 2006 Apr 18;78(S1):S22-S23
Evaluation of FDA licensed HIV assays using plasma from Cameroonian blood donors.
Lee S, Hu J, Tang S, Wood O, Francis K, Machuca A, Rios M, Daniel S, Vockley C, Awazi B, Zekeng L, Hewlett I

J Infect Dis 2006 Feb 1;193(3):427-37
Human T Lymphotropic Virus Types I and II Western Blot Seroindeterminate Status and Its Association with Exposure to Prototype HTLV-I.
Yao K, Hisada M, Maloney E, Yamano Y, Hanchard B, Wilks R, Rios M, Jacobson S

Clin Infect Dis 2005 Jun 1;40(11):1673-6
Seroprevalence of human T cell leukemia virus in HIV antibody-negative populations in rural Cameroon.
Machuca A, Wood O, Lee S, Daniel S, Rios M, Wolfe ND, Carr JK, Eitel MN, Tamoufe U, Torimiro JN, Burke D, Hewlett IK

Transfus Med 2005 Feb;15(1):49-55.
High frequency of partial DIIIa and DAR alleles found in sickle cell disease patients suggests increased risk of alloimmunization to RhD.
Castilho L, Rios M, Rodrigues A, Pellegrino J Jr, Saad ST, Costa FF

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