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Journal of Clinical Microbiology, July 2000, p. 2772-2773, Vol. 38, No. 7
Department of
Microbiology1 and Department of
Anatomical and Cellular Pathology,2 The Chinese
University of Hong Kong, Prince of Wales Hospital, Shatin, Hong
Kong SAR, China
Received 6 March 2000/Returned for modification 27 March
2000/Accepted 28 April 2000
Brain tissues from both sides of the cerebellum and frontal,
temporal, parietal, and occipital lobes were collected postmortem from
30 patients for human herpesvirus 8 (HHV-8) detection by PCR. Overall,
42 of 300 (14.0%) samples were positive, with similar rates for each
position. Nineteen patients (63.3%) were positive and showed a
significant increase in positivity with age (P = 0.007). The results indicate a neuroinvasive and neuropersistent potential of HHV-8.
Human herpesvirus 8 (HHV-8), also
known as Kaposi's sarcoma (KS)-associated herpesvirus, is a new member
of the Rhadinovirus genus in the
Gammaherpesvirinae subfamily (5). HHV-8 is
associated with all forms of KS in both immunocompromised and
immunocompetent patients (2, 13). The virus has been found
in primary effusion lymphoma (4, 11) and multicentric
Castleman's disease (7). HHV-8 DNA can also be detected in
noninvolved tissues from KS patients, including skin, peripheral blood
mononuclear cells, lymphoid tissue, prostate, and semen (10, 14,
15). While the spectrum of in vivo tropism of HHV-8 has yet to be
fully elucidated, reports of detection of HHV-8 in dorsal root ganglia
of KS patients (6), cerebrospinal fluid from human
immunodeficiency virus-positive patients (3), and brain
biopsy specimens from patients with unexplained encephalitis
(12) suggest a neuroinvasive potential of this novel virus.
In this study, the prevalence and distribution of HHV-8 in brain were examined.
Study samples.
Thirty consecutive unselected postmortem cases
were included. For each patient, a fresh autopsy sample was collected
from the cerebellum and frontal, temporal, parietal, and occipital lobes of both sides of the brain. Heart and kidney tissues of the same
patient were also collected as controls. Hemorrhagic areas and areas
with visible blood vessels were excluded. The study was approved by the
local institutional ethics committee.
HHV-8 detection.
DNA was extracted from a 2-mm3
tissue block by the QIAamp Tissue Kit (Qiagen, Hilden, Germany), with
the quality confirmed by a single-round PCR using primers PC03-PC07
targeting a 355-bp fragment of the
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Copyright © 2000, American Society for Microbiology. All rights reserved.
Survey for the Presence and Distribution of Human Herpesvirus
8 in Healthy Brain
ABSTRACT
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-globin gene as previously
described (9). HHV-8 DNA was detected by a nested PCR (outer
primers KS-1-KS-2, 5'-AGC CGA AAG GAT TCC ACC AT-3' and
5'-TTC GTG TTG TCT ACG TCC AG-3'; inner primers H8NS1-H8NS2,
5'-ACG GAT TTG ACC CCG TGT TC-3' and 5'-AAT GAC ACA TTG GTG
GTA TA-3') targeting a 233-bp and a 160-bp fragment,
respectively, of open reading frame (ORF) 26 (5, 10). Five
microliters of extracted DNA was amplified in a 50-µl reaction
mixture containing PCR buffer (10 mM Tris-HCl [pH 8.3], 50 mM KCl,
and 1.5 mM MgCl2), 200 µM (each) deoxynucleoside triphosphates, 1 U of Taq polymerase (Pharmacia Biotech,
Uppsala, Sweden), and 0.25 µM (each) outer primers. The cycling
conditions were initial denaturation (94°C, 4 min); 30 cycles of
denaturation (94°C, 1 min), annealing (55°C, 1 min), and extension
(72°C, 1 min); and a final extension (72°C, 8 min). Two microliters
of the first-round amplicons was amplified in a second-round PCR with
the same conditions using inner primers, 55°C for annealing, and
omission of initial denaturation. PCR amplicons were electrophoresed and visualized by ethidium bromide staining. All reactions were carried
out under stringent conditions to avoid cross-contamination (8). A negative control was included following each fifth sample.
Statistical analysis. The chi-square test or Fisher's exact test was used to analyze categorical variables. The independent-sample t test was used for numerical variables. Two-tailed P values of <0.05 were regarded as significant.
The 30 studied patients (29 Chinese and 1 Portuguese) were aged 20 to 95 years (mean, 61.4; standard deviation, 20.2) with a male/female ratio of 2:1. Three patients died of intracranial hemorrhage, with one due to trauma and two from rupture of intracranial arterial aneurysms. The deaths of remaining patients were not related to the central nervous system. None of the patients had clinical or pathological findings suggestive of current viral infection or skin lesions compatible with KS. All extracted DNAs were positive by the-globin PCR. All PCR
controls, including heart and kidney tissues, were negative in the
HHV-8 PCR. Samples positive by the first set of HHV-8 PCR (ORF 26) were
all confirmed by the second set of HHV-8 PCR (ORF 25).
Overall, 300 brain tissue samples were examined with 42 (14.0%) being
positive for HHV-8 DNA. The positive rates for each anatomical position
were similar (Table 1). Nineteen patients (63.3%) had HHV-8 DNA detected, of which 10 (52.6%) harbored viral DNA at more than one anatomical position, including five patients with
two positive samples, one patient with three positive samples, and
three patients with four positive samples. A 78-year-old male patient
who died of hemothorax due to traumatic injury harbored viral DNA at
eight anatomical sites. The viral DNA-positive and -negative groups
showed no significant difference in sex distribution (male/female
ratio, 11:8 versus 9:2; P = 0.246 by Fisher's exact test). Patients positive for HHV-8 were significantly older than the
negative group (mean age, 68.2 versus 49.6 years; P = 0.012; 95% confidence interval, 4.32 to 32.73 by t test). A
significant trend of increase in positive rate with age was also
observed (age < 30 years, 0%; 30 to 49 years, 42.9%; 50 to 69 years,
63.6%; >69 years, 90.0%; 
2 for linear trend, 7.336;
P = 0.007).
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FOOTNOTES |
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* Corresponding author. Mailing address: Department of Microbiology, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong SAR, China. Phone: (852) 2632-3333. Fax: (852) 2647-3227. E-mail: paulkschan{at}cuhk.edu.hk.
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REFERENCES |
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Text
References
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| 1. | Boshoff, C., S. Talbot, M. Kennedy, J. O'Leary, T. Schulz, and Y. Chang. 1996. HHV8 and skin cancers in immunosuppressed patients. Lancet 348:138[Medline]. |
| 2. | Boshoff, C., D. Whitby, T. Hatziioannou, C. Fisher, W. J. van der, A. Hatzakis, R. Weiss, and T. Schulz. 1995. Kaposi's-sarcoma-associated herpesvirus in HIV-negative Kaposi's sarcoma. Lancet 345:1043-1044[Medline]. |
| 3. |
Brink, N. S.,
Y. Sharvell,
M. R. Howard,
J. D. Fox,
M. J. Harrison, and R. F. Miller.
1998.
Detection of Epstein-Barr virus and Kaposi's sarcoma-associated herpesvirus DNA in CSF from persons infected with HIV who had neurological disease.
J. Neurol. Neurosurg. Psychiatry
65:191-195 |
| 4. |
Cesarman, E.,
Y. Chang,
P. S. Moore,
J. W. Said, and D. M. Knowles.
1995.
Kaposi's sarcoma-associated herpesvirus-like DNA sequences in AIDS-related body-cavity-based lymphomas.
N. Engl. J. Med.
332:1186-1191 |
| 5. |
Chang, Y.,
E. Cesarman,
M. S. Pessin,
F. Lee,
J. Culpepper,
D. M. Knowles, and P. S. Moore.
1994.
Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi's sarcoma.
Science
266:1865-1869 |
| 6. |
Corbellino, M.,
C. Parravicini,
J. T. Aubin, and E. Berti.
1996.
Kaposi's sarcoma and herpesvirus-like DNA sequences in sensory ganglia.
N. Engl. J. Med.
334:1341-1342 |
| 7. |
Gessain, A.,
A. Sudaka,
J. Briere,
N. Fouchard,
M. A. Nicola,
B. Rio,
M. Arborio,
X. Troussard,
J. Audouin, and J. Diebold.
1996.
Kaposi sarcoma-associated herpes-like virus (human herpesvirus type 8) DNA sequences in multicentric Castleman's disease: is there any relevant association in non-human immunodeficiency virus-infected patients?
Blood
87:414-416 |
| 8. | Kwok, S., and R. Higuchi. 1989. Avoiding false positives with PCR. Nature 339:237-238[CrossRef][Medline]. |
| 9. |
Lo, Y. M.,
W. Z. Mehal, and K. A. Fleming.
1989.
In vitro amplification of hepatitis B virus sequences from liver tumour DNA and from paraffin wax embedded tissues using the polymerase chain reaction.
J. Clin. Pathol.
42:840-846 |
| 10. |
Monini, P.,
L. de Lellis,
M. Fabris,
F. Rigolin, and E. Cassai.
1996.
Kaposi's sarcoma-associated herpesvirus DNA sequences in prostate tissue and human semen.
N. Engl. J. Med.
334:1168-1172 |
| 11. |
Nador, R. G.,
E. Cesarman,
D. M. Knowles, and J. W. Said.
1995.
Herpes-like DNA sequences in a body-cavity-based lymphoma in an HIV-negative patient.
N. Engl. J. Med.
333:943 |
| 12. | Said, J. W., T. Tasaka, S. de Vos, and H. P. Koeffler. 1997. Kaposi's sarcoma-associated herpesvirus/human herpesvirus type 8 encephalitis in HIV-positive and -negative individuals. AIDS 11:1119-1122[CrossRef][Medline]. |
| 13. | Schalling, M., M. Ekman, E. E. Kaaya, A. Linde, and P. Biberfeld. 1995. A role for a new herpes virus (KSHV) in different forms of Kaposi's sarcoma. Nat. Med. 1:707-708[CrossRef][Medline]. |
| 14. | Viviano, E., F. Vitale, F. Ajello, A. M. Perna, M. R. Villafrate, F. Bonura, M. Arico, G. Mazzola, and N. Romano. 1997. Human herpesvirus type 8 DNA sequences in biological samples of HIV-positive and negative individuals in Sicily. AIDS 11:607-612[CrossRef][Medline]. |
| 15. | Whitby, D., M. R. Howard, M. Tenant-Flowers, N. S. Brink, A. Copas, C. Boshoff, T. Hatzioannou, F. E. Suggett, D. M. Aldam, and A. S. Denton. 1995. Detection of Kaposi sarcoma associated herpesvirus in peripheral blood of HIV-infected individuals and progression to Kaposi's sarcoma. Lancet 346:799-802[CrossRef][Medline]. |
Journal of Clinical Microbiology, July 2000, p. 2772-2773, Vol. 38, No. 7
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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