Detection of herpes simplex virus DNA in donor cornea culture medium by polymerase chain reaction.

Morris, David J.; Cleator, Graham M.; Klapper, Paul E.; Cooper, Robert J.; Biney, Emmanuel O.E.; Dennett, C; Marcyniuk, B.; Tullo, Andrew B.

doi:10.1136/bjo.80.7.654

Cited by 42 publications

(33 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Only 3.8% of 80 corneas were positive for HSV and post transplant follow up of the three recipients revealed no evidence of HSV induced eye disease or primary graft failure (Morris et al 1996). In our study we could not demonstrate HSV DNA by PCR in any of the recipient remnants.…”

Section: Discussioncontrasting

confidence: 73%

The relationship of graft survival and herpes simplex virus latency in recipient corneal buttons

Aydemir

Türkçüoğlu²,

Bulut³

et al. 2007

OPTH

View full text Add to dashboard Cite

Purpose: To demonstrate relationship between herpes simplex virus (HSV) corneal latency and graft survival. Methods: Prospective case control study. 28 recipient corneal buttons and donor cornea-scleral remnants were examined for HSV DNA with polymerase chain reaction (PCR). None of the recipient had a history of HSV infection. Serum samples of graft recipients were analyzed for the presence of anti-HSV IgG and IgM with enzyme-linked immunosorbent assay technique. All corneas were free of stromal scarring or epithelial defect before sampling and had an endothelial cell density of >2000 cells/mm 2 . Results: In twenty three patients (82%) anti-HSV IgG was detected in serum. In none of the recipients anti-HSV IgM was positive. HSV DNA was positive in six out of twenty eight (21%) of the recipient corneal buttons and none of the donor cornea-scleral remnants. In eighteenmonths follow-up period three out of six (50%) HSV DNA positive and one out of twenty-two (4.5%) HSV DNA negative patients experienced late endothelial failure that was statistically signifi cantly different (p = 0.022). Conclusion: Even without a history of HSV keratitis, presence of latent HSV virus in recipient cornea is an important risk factor for subsequent graft survival.

show abstract

Section: Discussioncontrasting

confidence: 73%

The relationship of graft survival and herpes simplex virus latency in recipient corneal buttons

Aydemir

Türkçüoğlu²,

Bulut³

et al. 2007

OPTH

View full text Add to dashboard Cite

show abstract

“…The stability of DNA in culture media has important implications for all investigations in which the concentration of target DNA is low or in which maintenance of its integrity influences the outcome of results. Regarding this point, the study of Morris et al [1996] lacks information about storage conditions and the time from end of culture to extraction of DNA. These probably were not documented because the relevance of these factors has not been recognized.…”

Section: Discussionmentioning

confidence: 99%

“…Further evidence for such possibility comes from animal studies [Cook and Brown, 1986;Cantin et al, 1992]. Although surgical trauma, such as penetrating keratoplasty, may induce viral reactivation in cases of latent corneal infection [Nicholls et al, 1996], the possibility of donor-to-recipient transmission of HSV-1 DNA via transplanted corneae, and its potential pathogenic consequences, have not been investigated conclusively [Morris et al, 1996].…”

Section: Introductionmentioning

confidence: 99%

“…Analysis of culture medium samples for the presence of viral DNA is naturally undertaken on the presupposition that if present, this material has remained stable under the storage conditions adopted prior to use [Morris et al, 1996]; but this is an assumption which lacks experimental corroboration. The consideration is an important one since, if degraded, viral DNA will elude detection, thereby giving rise to a false negative result.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Low rate shedding of HSV-1 DNA, but not of infectious virus from human donor corneae into culture media

Garweg

Boehnke

1997

J. Med. Virol.

View full text Add to dashboard Cite

Fluid samples derived from 451 organ cultured corneae were tested for the presence of HSV-1 DNA after electroseparation and amplification for fragments of the glycoprotein D- and thymidine kinase-encoding genes. Of the culture media, 134 were processed immediately after withdrawal (Group 1); 100 were stored at ambient temperature for 6 to 60 weeks (Group 2); 90 were stored at -8 degrees C for 4 to 9 weeks (Group 3); and 127 were stored at -20 degrees C for 2 to 30 weeks (Group 4). The degradation of human DNA (marker gene, betaglobin) under these different storage conditions and of human and HSV-1 DNA as a sequential function of time at ambient temperature was gauged by the loss of a detectable signal for the respective component. Endothelial cell density within each of the corneal discs was determined before and after organ culture. In 7/451 culture fluid samples, HSV-1 DNA corresponding to either the glycoprotein D- or thymidine kinase-encoding genes was detected. In culture fluid samples derived from Group 2 at ambient temperature, for 6 to 60 weeks) and 3 (at -8 degrees C, for 4 to 9 weeks), complete degradation precluded the detection of human DNA, and hence probably also of HSV-1 DNA; only at -20 degrees C did DNA remain stable for protracted periods of time. Even so, HSV-1 DNA was detected in only 2% of those media in which no degradation was to be expected; additionally, there existed no correlation between its presence in culture fluid samples and the loss of endothelial cells or cytopathic changes. DNA can be extracted successfully and concentrated twenty-fold from high-volume samples by electroseparation. When shed into culture fluid, it is remarkably prone to a time and temperature dependent degradation, which may lead to false negative results. It is concluded that there is no infectious virus to be expected in the specimens; the occurrence of HSV-1 DNA in donor corneae would not appear to be an important factor influencing their biological quality during the period of organ culture.

show abstract

“…A typical dendritic keratitis was found on a cultured cornea [8] , and HSV-1 DNA was detected by PCR in 3 cultured corneas with endothelial necrosis [9] . Also, 2 reports have shown that HSV-1 DNA can be detected in the preservation medium: the first study showed HSV-1 DNA in 3 cell pellets out of 80 culture media [10] , and the second revealed 12 media contaminated by HSV-1 DNA in a series of 199 corneas with severe or complete endothelial necrosis [11] . Finally, Garweg and Boehnke [12] demonstrated a time-and temperature-dependent degradation of HSV-1 DNA once introduced in the culture medium, in a series of 451 corneal preservation media with different conditions of preservation (31, 4, -20 and -80 ° C).…”

Section: Introductionmentioning

confidence: 99%

The Survival of Herpes Simplex Virus in Preserved Murine Corneas

Robert¹,

Ducher

Pleyer

et al. 2008

Ophthalmic Res

View full text Add to dashboard Cite

Background/Aims: This study investigates the survival of HSV in infected mouse corneas, in the conditions of normal human eye bank preservation. Methods: Hundred seventy-two BALB/C mice infected with herpes simplex virus 1 (HSV-1) (KOS) were randomly assigned to either: no preservation in group 1 (n = 62), 31°C preservation for 3 weeks in group 2 (n = 70) or 4°C preservation for 8 days in group 3 (n = 40). The presence of HSV-1 was thereafter detected by viral culture and PCR. Results: In groups 1, 2 and 3, HSV-1 was detected by culture in 22 (35.5%), 1 (1.4%) and 0 (0.0%) of the corneas, and by PCR in 27 (43.7%), 3 (4.2%) and 7 (17.5%) of the corneas respectively. When compared to group 1, HSV was detected significantly less often in groups 2 (p < 0.0001) and 3 (p < 0.0001). Conclusions: HSV-1 DNA undergoes a degradation during corneal preservation.

show abstract

Detection of herpes simplex virus DNA in donor cornea culture medium by polymerase chain reaction.

Cited by 42 publications

References 18 publications

The relationship of graft survival and herpes simplex virus latency in recipient corneal buttons

The relationship of graft survival and herpes simplex virus latency in recipient corneal buttons

Low rate shedding of HSV-1 DNA, but not of infectious virus from human donor corneae into culture media

The Survival of Herpes Simplex Virus in Preserved Murine Corneas

Contact Info

Product

Resources

About