A new arenavirus in a cluster of fatal transplant-associated diseases.
BACKGROUND
Three patients who received visceral-organ transplants from a single donor on the same day died of a febrile illness 4 to 6 weeks after transplantation. Culture, polymerase-chain-reaction (PCR) and serologic assays, and oligonucleotide microarray analysis for a wide range of infectious agents were not informative.


METHODS
We evaluated RNA obtained from the liver and kidney transplant recipients. Unbiased high-throughput sequencing was used to identify microbial sequences not found by means of other methods. The specificity of sequences for a new candidate pathogen was confirmed by means of culture and by means of PCR, immunohistochemical, and serologic analyses.


RESULTS
High-throughput sequencing yielded 103,632 sequences, of which 14 represented an Old World arenavirus. Additional sequence analysis showed that this new arenavirus was related to lymphocytic choriomeningitis viruses. Specific PCR assays based on a unique sequence confirmed the presence of the virus in the kidneys, liver, blood, and cerebrospinal fluid of the recipients. Immunohistochemical analysis revealed arenavirus antigen in the liver and kidney transplants in the recipients. IgM and IgG antiviral antibodies were detected in the serum of the donor. Seroconversion was evident in serum specimens obtained from one recipient at two time points.


CONCLUSIONS
Unbiased high-throughput sequencing is a powerful tool for the discovery of pathogens. The use of this method during an outbreak of disease facilitated the identification of a new arenavirus transmitted through solid-organ transplantation.
Palacios
Gustavo
G
Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, NY 10032, USA.
Druce
Julian
J
Du
Lei
L
Tran
Thomas
T
Birch
Chris
C
Briese
Thomas
T
Conlan
Sean
S
Quan
Phenix-Lan
PL
Hui
Jeffrey
J
Marshall
John
J
Simons
Jan Fredrik
JF
Egholm
Michael
M
Paddock
Christopher D
CD
Shieh
Wun-Ju
WJ
Goldsmith
Cynthia S
CS
Zaki
Sherif R
SR
Catton
Mike
M
Lipkin
W Ian
WI
D000328:Adult; D000914:Antibodies, Viral / Q000097:blood; D001117:Arenaviridae Infections / Q000635:transmission / Q000821:virology*; D018050:Arenavirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D019295:Computational Biology; D018562:Disease Transmission, Infectious; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007668:Kidney / Q000648:ultrastructure / Q000821:virology; D016030:Kidney Transplantation / Q000009:adverse effects*; D016031:Liver Transplantation / Q000009:adverse effects*; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction / Q000379:methods; D012367:RNA, Viral / Q000032:analysis; D017422:Sequence Analysis, DNA / Q000379:methods*
D000914:Antibodies, Viral; D012367:RNA, Viral