Concepcion Conejero-Goldberg1, Ena Wang2, Chuli Yi1,

Terry Goldberg3, Francesco M. Marincola2, Lorraine

Jones-Brando4, Maree J. Webster1, Robert H. Yolken4,

and E. Fuller Torrey1



Medical Research Institute, 2Department of Transfusion Medicine,

National Institutes of Health, 3Clinical Brain Disorders Branch,

National Institutes of Health, 4Stanley Division of Developmental

Neurovirology, Johns Hopkins University School of Medicine




Microarray technology has become an important tool in psychiatric research.  A

unique array based pathogen chip has been developed in our laboratory for the

detection of viral RNA expression levels or DNA prevalence from test samples.  A

set of long oligonucleotides (60-mer) was designed based on highly conserved

regions within viral families, as well as heterogenic regions characterized by

individual subfamilies.  In addition, by including oligonucleotides derived from

genes implicated in different stages of the infection, we were also able to

potentially define the stage of the viral infection.  To validate the viral

microarray we used virtually infected cell cultures to detect and identify

diverse viruses and their infectious stage.


Total RNA

from Brodmann area 46, from 8 patients with schizophrenia and 10 unaffected

controls was linearly amplified using in vitro transcription in combination with

a template switch technique.  Samples were then examined by cohybridization of

patient aRNA labeled with Cy5 (red) with pooled control aRNA labeled with Cy3

(green) to the pathogen microarray chip.  The expression of 51 genes displayed

statistically significant differences between schizophrenic cases and controls

(P<.05 by t-test).  Expression of 6 viral sequences differed between groups by X2

(using a 1 SD cut off).  All favored increased levels of expression in the

schizophrenia cases.  We found evidence of increased expression of sequences

homologous to retroviruses, human cytomegalovirus and HSV-2 in the schizophrenic

group.  Quantitative-PCR studies are underway to validate these results.



findings suggest that this platform provides the capability to detect a broad

spectrum of viruses in a single array while simultaneously discriminating among

different stages of the viruses.  This method may be applied to identify

evidence of viral infection in postmortem tissue from psychiatric patients as

well as a wide range of other diagnostic categories.