Serology in the Digital Age: Using Long Synthetic Peptides Created from Nucleic Acid Sequences as Antigens in Microarrays.
BACKGROUND
Antibodies to microbes, or to autoantigens, are important markers of disease. Antibody detection (serology) can reveal both past and recent infections. There is a great need for development of rational ways of detecting and quantifying antibodies, both for humans and animals. Traditionally, serology using synthetic antigens covers linear epitopes using up to 30 amino acid peptides.


METHODS
We here report that peptides of 100 amino acids or longer ("megapeptides"), designed and synthesized for optimal serological performance, can successfully be used as detection antigens in a suspension multiplex immunoassay (SMIA). Megapeptides can quickly be created just from pathogen sequences. A combination of rational sequencing and bioinformatic routines for definition of diagnostically-relevant antigens can, thus, rapidly yield efficient serological diagnostic tools for an emerging infectious pathogen.


RESULTS
We designed megapeptides using bioinformatics and viral genome sequences. These long peptides were tested as antigens for the presence of antibodies in human serum to the filo-, herpes-, and polyoma virus families in a multiplex microarray system. All of these virus families contain recently discovered or emerging infectious viruses.


CONCLUSIONS
Long synthetic peptides can be useful as serological diagnostic antigens, serving as biomarkers, in suspension microarrays.
Section of Clinical Virology, Department of Medical Sciences, Uppsala University, Uppsala 751 85, Sweden. rizwan.muhd715@gmail.com.|Laboratory of Clinical Microbiology, Uppsala University Hospital, Uppsala 751 85, Sweden. rizwan.muhd715@gmail.com.
Muhammad
M
Rizwan
Section of Clinical Virology, Department of Medical Sciences, Uppsala University, Uppsala 751 85, Sweden. bengt.ronnberg@gmail.com.|Laboratory of Clinical Microbiology, Uppsala University Hospital, Uppsala 751 85, Sweden. bengt.ronnberg@gmail.com.
Bengt
B
Rönnberg
Section of Clinical Virology, Department of Medical Sciences, Uppsala University, Uppsala 751 85, Sweden. Maksims.Cistjakovs@rsu.lv.|August Kirchenstein Institute of Microbiology and Virology, Riga Stradins University, Riga LV-1067, Latvia. Maksims.Cistjakovs@rsu.lv.
Maksims
M
Cistjakovs
Laboratory of Clinical Microbiology, Uppsala University Hospital, Uppsala 751 85, Sweden. ake.lundkvist@imbim.uu.se.|Zoonosis Science Center, Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala 75123, Sweden. ake.lundkvist@imbim.uu.se.
Åke
Å
Lundkvist
Deutsches Krebsforschungszentrum, Heidelberg 69121, Germany. pipkorn@t-online.de.
Rudiger
R
Pipkorn
Section of Clinical Virology, Department of Medical Sciences, Uppsala University, Uppsala 751 85, Sweden. Jonas.blomberg@medsci.uu.se.|Zoonosis Science Center, Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala 75123, Sweden. Jonas.blomberg@medsci.uu.se.|Laboratory of Clinical Microbiology, Linköping University Hospital, Linköping 58185, Sweden. Jonas.blomberg@medsci.uu.se.
Jonas
J
Blomberg
emerging virus infections; immunoassay; megapeptide; suspension microarray; suspension multiplex immunoassay (SMIA)