Detection of Viral DNA in Neonatal Herpes Encephalitis Autopsy Tissues by Solution‐Phase PCR: Comparison with Pathology and Immunohistochemistry

Gressèns, Pierre; Langston, Claire; Mitchell, William J.; Martin, John R.

doi:10.1111/j.1750-3639.1993.tb00750.x

Cited by 15 publications

(5 citation statements)

References 36 publications

(31 reference statements)

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“…Furthermore, only lungs five days post-infection were positive by immunohistochemistry. This can be explained by the higher sensitivity normally shown in PCR reactions when compared to immunohistochemistry, as observed by other authors in the detection of several infectious agents, including viruses [6,13,14,26]. By immunohistochemistry the presence of the BRSV antigen in bronchial epithelial cells, as detected by Prince et al [22] in cotton rats infected with HRSV, was verified.…”

Section: Discussionmentioning

confidence: 76%

Detection of bovine respiratory syncytial virus in experimentally infected balb/c mice

et al. 2004

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-The present study used an RT-nested-PCR and an immunohistochemistry assay to detect bovine respiratory syncytial virus in tissues from experimentally infected balb/c mice. As a first step, Chicken Embryo Related (CER) cell monolayers infected with the BRSV-25-BR strain isolated in Brazil were used for antigen production. Then, the infected lung and tracheal tissues of female balb/c mice were collected on 3, 5, 7 and 10 days post-infection and submitted to both techniques. Primers specific to F and G genes that amplify fragments of 481 bp and 371 bp, respectively, were used. The BRSV detection was not successful in all of the animals tested. The

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Section: Discussionmentioning

confidence: 76%

Detection of bovine respiratory syncytial virus in experimentally infected balb/c mice

et al. 2004

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“…The diagnostic value of post-mortem specimens for this purpose is well described [27-33]. Autopsies are routinely conducted in Bangladesh as a part of police investigations into possible homicides, suicides, or other unnatural deaths but not outside of this context [34].…”

Section: Introductionmentioning

confidence: 99%

Family and community concerns about post-mortem needle biopsies in a Muslim society

et al. 2011

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BackgroundPost-mortem needle biopsies have been used in resource-poor settings to determine cause of death and there is interest in using them in Bangladesh. However, we did not know how families and communities would perceive this procedure or how they would decide whether or not to consent to a post-mortem needle biopsy. The goal of this study was to better understand family and community concerns and decision-making about post-mortem needle biopsies in this low-income, predominantly Muslim country in order to design an informed consent process.MethodsWe conducted 16 group discussions with family members of persons who died during an outbreak of Nipah virus illness during 2004-2008 and 11 key informant interviews with their community and religious leaders. Qualitative researchers first described the post-mortem needle biopsy procedure and asked participants whether they would have agreed to this procedure during the outbreak. Researchers probed participants about the circumstances under which the procedure would be acceptable, if any, their concerns about the procedure, and how they would decide whether or not to consent to the procedure.ResultsOverall, most participants agreed that post-mortem needle biopsies would be acceptable in some situations, particularly if they benefitted society. This procedure was deemed more acceptable than full autopsy because it would not require major delays in burial or remove organs, and did not require cutting or stitching of the body. It could be performed before the ritual bathing of the body in either the community or hospital setting. However, before consent would be granted for such a procedure, the research team must gain the trust of the family and community which could be difficult. Although consent may only be provided by the guardians of the body, decisions about consent for the procedure would involve extended family and community and religious leaders.ConclusionsThe possible acceptability of this procedure during outbreaks represents an important opportunity to better characterize cause of death in Bangladesh which could lead to improved public health interventions to prevent these deaths. Obstacles for research teams will include engaging all major stakeholders in decision-making and quickly building a trusting relationship with the family and community, which will be difficult given the short window of time prior to the ritual bathing of the body.

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“…DNA samples were prepared from frozen corneas (Mitchell & Martin, 1992) or from paraffinembedded sections of cornea (Gressens et al, 1993). Approximately 100 ng of DNA from frozen corneas was added to 200 laM each dNTP, 1.5 mM-MgCI~, 50 mM-KC1; 10 mM-Tris-HCl pH 8.3, 0-001% gelatin, 2-5 U of Taq polymerase and 100 ng of each primer and subjected to 30 cycles of PCR.…”

Section: Methodsmentioning

confidence: 99%

Herpes simplex virus type 1 DNA persistence, progressive disease and transgenic immediate early gene promoter activity in chronic corneal infections in mice

Mitchell¹,

Gressèns²,

Martin³

et al. 1994

Journal of General Virology

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We have used a mouse model system and the corneal route of inoculation to examine the issue of extraneuronal persistence of herpes simplex virus type 1 (HSV-1). HSV-1 strain F DNA and inflammatory lesions were detected in corneal tissue of mice at 5, 11, 23, 37 and 60 days post-infection (p.i.). Viral DNA was localized by in situ PCR to epithelial cells and less frequently to cells in the stroma of the cornea. Viral proteins were not detected in the cornea and virus could not be isolated from tissue homogenates after 11 days p.i. even though histopathological lesions became progressively more severe at 37 and 60 days p.i. The DNA-containing cells were usually adjacent to the sites of inflammation or within these sites in the chronic stage (23, 37 and 60 days p.i.). In contrast to strain F, persistence of HSV-1 strain KOS DNA and inflammatory lesions were not detected after 11 days p.i. ; this result suggests that the long-term persistence of HSV-1 DNA and the development of inflammatory lesions are virus strain-dependent. We tested for the possibility of transgenic HSV-1 immediate early gene (ICP4) promoter activity in chronically infected corneas of transgenic mice containing the ICP4 promoter fused to the bacterial fl-galactosidase coding sequence. Our results indicated that the chimeric transgene was expressed in the cornea at 5, 11, 23, 37 and 41 days p.i. Possible explanations for these results and mechanisms for the generation of the chronic inflammatory lesions are discussed. The properties of chronic HSV infections in the cornea may be similar to those which have been described for persistent or defective viral infections in other systems.

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Detection of Viral DNA in Neonatal Herpes Encephalitis Autopsy Tissues by Solution‐Phase PCR: Comparison with Pathology and Immunohistochemistry

Cited by 15 publications

References 36 publications

Detection of bovine respiratory syncytial virus in experimentally infected balb/c mice

Detection of bovine respiratory syncytial virus in experimentally infected balb/c mice

Family and community concerns about post-mortem needle biopsies in a Muslim society

Herpes simplex virus type 1 DNA persistence, progressive disease and transgenic immediate early gene promoter activity in chronic corneal infections in mice

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