Failure to Detect Borna Disease Virus p24 and p40 mRNA in 60 Autopsy Brain Samples of the Stanley Foundation Brain Collection

FAILURE

TO DETECT BORNA DISEASE VIRUS p24 AND P40 mrna IN 60 AUTOPSY

BRAIN SAMPLES OF THE STANLEY FOUNDATION BRAIN COLLECTION

K. Ikeda*, S.

Haga, Y.Motoi, T. Aizawa, K. Arima. Department of Ultrastructure

and Histochemistry, Tokyo Institute of Psychiatry, Tokyo, Japan

There is evidence suggesting an

association of Borna Disease Virus (BDV) with certain psychiatric

disorders. To address the issues, several studies have been done

for detecting BDV-specific mRNA in autopsy brain tissues of

individuals with psychiatric disorders and controls. Thus far,

contradictory reports have been presented, covering the report of

the lack of detection and the report of very high prevalence of

BDV mRNA in samples from people with schizophrenia. We conducted

a similar study by nested RT-PCR for the brain samples of high

quality, with special precautions for both the false-positivity

and false-negativity in mind.

All the coded, autopsy brain

samples were provided from the Stanley Foundation Neuropathology

Consortium; tissues of each frontal and temporal cortex of 45

individuals with schizophrenia (15), bipolar disorder without

psychosis (15), and depression (15), and 15 controls. Total RNA

was extracted with Qiagen kit. To avoid the laboratory

contamination of BDV related nucleic acids, special cares were

paid for the processes of RNA extraction and the nested RT-PCR

assay. The RNA transcribed from the plasmid containing BDV p24 or

p40 sequence with an inserted foreign oligonucleotide (~50 nt),

P24 +@ or p40 + @ , was used as a positive control.

All the RNA samples of brain

tissues were capable to be amplified by nested RT-PCR for G3PDH

mRNA. Our nested RT-PCR assay detected reproducibly 500 copies of

p24+@ RNA and 100 copies of p40+@ RNA, showing the high sensitivity for detecting

BDV p24 or p40. No PCR products for p24 or p40 were observed in

any of the 120 samples.

The previous data including ours

on the detection of BDV in autopsy brain samples should be

reevaluated by the sensitive and careful nested RT-PCR assay.