VIRAL DETECTION ARRAYS

Concepcion Conejero-Goldberg¹, Ena Wang², Chuli Yi¹,

Terry Goldberg³, Francesco M. Marincola², Lorraine

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

1Stanley

Medical Research Institute, ²Department of Transfusion Medicine,

National Institutes of Health, ³Clinical Brain Disorders Branch,

National Institutes of Health, ⁴Stanley Division of Developmental

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

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

These

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