King's College London

Research portal

Detection of viral DNA in neonatal herpes encephalitis autopsy tissues by solution-phase PCR: comparison with pathology and immunohistochemistry

Research output: Contribution to journalArticle

Standard

Detection of viral DNA in neonatal herpes encephalitis autopsy tissues by solution-phase PCR : comparison with pathology and immunohistochemistry. / Gressens, P; Langston, C; Mitchell, W J; Martin, J R; Gressens, Pierre.

In: Brain Pathology, Vol. 3, No. 3, 07.1993, p. 237-50.

Research output: Contribution to journalArticle

Harvard

Gressens, P, Langston, C, Mitchell, WJ, Martin, JR & Gressens, P 1993, 'Detection of viral DNA in neonatal herpes encephalitis autopsy tissues by solution-phase PCR: comparison with pathology and immunohistochemistry', Brain Pathology, vol. 3, no. 3, pp. 237-50. https://doi.org/10.1111/j.1750-3639.1993.tb00750.x

APA

Gressens, P., Langston, C., Mitchell, W. J., Martin, J. R., & Gressens, P. (1993). Detection of viral DNA in neonatal herpes encephalitis autopsy tissues by solution-phase PCR: comparison with pathology and immunohistochemistry. Brain Pathology, 3(3), 237-50. https://doi.org/10.1111/j.1750-3639.1993.tb00750.x

Vancouver

Gressens P, Langston C, Mitchell WJ, Martin JR, Gressens P. Detection of viral DNA in neonatal herpes encephalitis autopsy tissues by solution-phase PCR: comparison with pathology and immunohistochemistry. Brain Pathology. 1993 Jul;3(3):237-50. https://doi.org/10.1111/j.1750-3639.1993.tb00750.x

Author

Gressens, P ; Langston, C ; Mitchell, W J ; Martin, J R ; Gressens, Pierre. / Detection of viral DNA in neonatal herpes encephalitis autopsy tissues by solution-phase PCR : comparison with pathology and immunohistochemistry. In: Brain Pathology. 1993 ; Vol. 3, No. 3. pp. 237-50.

Bibtex Download

@article{0ad81ce712be4a2094cbcdc8807092e6,
title = "Detection of viral DNA in neonatal herpes encephalitis autopsy tissues by solution-phase PCR: comparison with pathology and immunohistochemistry",
abstract = "To detect DNA sequences of herpes simplex virus (HSV) in neural and non-neural tissue sections in disseminated human neonatal HSV infection, a solution polymerase chain reaction (PCR) protocol was developed which amplified HSV thymidine kinase and host genomic DNA sequences that were hybridized with sequence-specific probes in Southern blots. Serial sections of formalin-fixed, paraffin embedded autopsy tissues were tested by PCR and compared to histology and HSV antigen detection. The sensitivity, specificity and reproducibility of this PCR protocol were determined on uninfected and HSV-infected mouse tissues and on HSV DNA from infected tissue culture cells. Samples estimated to contain as few as 60 copies of preserved HSV DNA target sequence gave a positive PCR result. In nine neonates that died during acute HSV infection, all non-neural tissues and a minority of neural tissues with histological lesions had HSV antigen; when DNA could be amplified, HSV DNA sequences were detected by PCR. Together, these findings indicate a direct role for virus in the pathogenesis of these lesions. In the same cases, some or all brain samples were negative for HSV antigen, but nevertheless had HSV DNA sequences detected by PCR. The possible explanations for this finding are discussed. In one neonate dying seven weeks after birth, HSV sequences were found in brain lesions in the absence of HSV antigen; neither HSV DNA nor antigen were found in non-neural tissues, suggesting a latent HSV infection in brain. It is practical to apply PCR methods to detect minute quantities of viral DNA in formalin-fixed, paraffin embedded autopsy tissues.(ABSTRACT TRUNCATED AT 250 WORDS)",
keywords = "Base Sequence, Blotting, Southern, DNA, Viral, Encephalitis, Female, Herpes Simplex, Humans, Immunohistochemistry, Infant, Infant, Newborn, Male, Molecular Sequence Data, Polymerase Chain Reaction, Simplexvirus, Thymidine Kinase",
author = "P Gressens and C Langston and Mitchell, {W J} and Martin, {J R} and Pierre Gressens",
year = "1993",
month = jul,
doi = "10.1111/j.1750-3639.1993.tb00750.x",
language = "English",
volume = "3",
pages = "237--50",
journal = "Brain Pathology",
issn = "1015-6305",
number = "3",

}

RIS (suitable for import to EndNote) Download

TY - JOUR

T1 - Detection of viral DNA in neonatal herpes encephalitis autopsy tissues by solution-phase PCR

T2 - comparison with pathology and immunohistochemistry

AU - Gressens, P

AU - Langston, C

AU - Mitchell, W J

AU - Martin, J R

AU - Gressens, Pierre

PY - 1993/7

Y1 - 1993/7

N2 - To detect DNA sequences of herpes simplex virus (HSV) in neural and non-neural tissue sections in disseminated human neonatal HSV infection, a solution polymerase chain reaction (PCR) protocol was developed which amplified HSV thymidine kinase and host genomic DNA sequences that were hybridized with sequence-specific probes in Southern blots. Serial sections of formalin-fixed, paraffin embedded autopsy tissues were tested by PCR and compared to histology and HSV antigen detection. The sensitivity, specificity and reproducibility of this PCR protocol were determined on uninfected and HSV-infected mouse tissues and on HSV DNA from infected tissue culture cells. Samples estimated to contain as few as 60 copies of preserved HSV DNA target sequence gave a positive PCR result. In nine neonates that died during acute HSV infection, all non-neural tissues and a minority of neural tissues with histological lesions had HSV antigen; when DNA could be amplified, HSV DNA sequences were detected by PCR. Together, these findings indicate a direct role for virus in the pathogenesis of these lesions. In the same cases, some or all brain samples were negative for HSV antigen, but nevertheless had HSV DNA sequences detected by PCR. The possible explanations for this finding are discussed. In one neonate dying seven weeks after birth, HSV sequences were found in brain lesions in the absence of HSV antigen; neither HSV DNA nor antigen were found in non-neural tissues, suggesting a latent HSV infection in brain. It is practical to apply PCR methods to detect minute quantities of viral DNA in formalin-fixed, paraffin embedded autopsy tissues.(ABSTRACT TRUNCATED AT 250 WORDS)

AB - To detect DNA sequences of herpes simplex virus (HSV) in neural and non-neural tissue sections in disseminated human neonatal HSV infection, a solution polymerase chain reaction (PCR) protocol was developed which amplified HSV thymidine kinase and host genomic DNA sequences that were hybridized with sequence-specific probes in Southern blots. Serial sections of formalin-fixed, paraffin embedded autopsy tissues were tested by PCR and compared to histology and HSV antigen detection. The sensitivity, specificity and reproducibility of this PCR protocol were determined on uninfected and HSV-infected mouse tissues and on HSV DNA from infected tissue culture cells. Samples estimated to contain as few as 60 copies of preserved HSV DNA target sequence gave a positive PCR result. In nine neonates that died during acute HSV infection, all non-neural tissues and a minority of neural tissues with histological lesions had HSV antigen; when DNA could be amplified, HSV DNA sequences were detected by PCR. Together, these findings indicate a direct role for virus in the pathogenesis of these lesions. In the same cases, some or all brain samples were negative for HSV antigen, but nevertheless had HSV DNA sequences detected by PCR. The possible explanations for this finding are discussed. In one neonate dying seven weeks after birth, HSV sequences were found in brain lesions in the absence of HSV antigen; neither HSV DNA nor antigen were found in non-neural tissues, suggesting a latent HSV infection in brain. It is practical to apply PCR methods to detect minute quantities of viral DNA in formalin-fixed, paraffin embedded autopsy tissues.(ABSTRACT TRUNCATED AT 250 WORDS)

KW - Base Sequence

KW - Blotting, Southern

KW - DNA, Viral

KW - Encephalitis

KW - Female

KW - Herpes Simplex

KW - Humans

KW - Immunohistochemistry

KW - Infant

KW - Infant, Newborn

KW - Male

KW - Molecular Sequence Data

KW - Polymerase Chain Reaction

KW - Simplexvirus

KW - Thymidine Kinase

U2 - 10.1111/j.1750-3639.1993.tb00750.x

DO - 10.1111/j.1750-3639.1993.tb00750.x

M3 - Article

C2 - 8293183

VL - 3

SP - 237

EP - 250

JO - Brain Pathology

JF - Brain Pathology

SN - 1015-6305

IS - 3

ER -

View graph of relations

© 2018 King's College London | Strand | London WC2R 2LS | England | United Kingdom | Tel +44 (0)20 7836 5454