Cytological Diagnosis of Herpesvirus Keratitis by Means of in situ Hybridization: Report of a Case

Kobayashi, Tadao K.; Sato, Shiroh; Iwa, Nobuzo; Yakushiji, Michiyasu

doi:10.1159/000310235

Cited by 6 publications

(4 citation statements)

References 7 publications

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“…The technique can also be applied to cytological specimens, [12][13][14] in some instances after destaining a Papanicolou preparation. 15 Most authors stress the importance of rapid fixation when RNA is to be hybridized, because of the potential for degradation by the ubiquitous RNases, and initial ISH studies indicated that delay in fixation results in loss of signal for mRNA. 6,16 However, there are now several reports which indicate that RNA can be extracted from post-mortem tissues [17][18][19] and that hybridization to mRNA 20,21 and viral target sequences 22,23 can be successfully detected in tissues unfixed for at least 24 h or obtained at autopsy.…”

Section: Preparation Of Tissuesmentioning

confidence: 99%

In Situ hybridization and its diagnostic applications in pathology

1997

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In situ hybridization (ISH) is a technique by which specific nucleotide sequences are identified in cells or tissue sections. These may be endogenous, bacterial or viral, DNA or RNA. On the basis of research applications, the technique is now being translated into diagnostic practice, mainly in the areas of gene expression, infection and interphase cytogenetics. Diagnostic applications are most often based on short nucleotide sequences (oligomers) labelled with non‐isotopic reporter molecules, and sites of binding may be localized by histochemical or immunohistochemical methods. The technique can be applied to routinely fixed and processed tissues; with some targets, it is even possible to obtain hybridization in autopsy material. ISH has been used to detect messenger RNA (mRNA) as a marker of gene expression, where levels of protein storage are low; for example, to confirm an endocrine tumour as the source of excess hormone production. Its application in infectious diseases has to date been mainly in viral infections, such as the typing of human papillomavirus (HPV) or the detection of Epstein–Barr virus by the presence of small nuclear RNAs (EBERs). The expression of mRNAs for histone proteins has been used to detect cells in S phase, and related methods may be applied to detect apoptotic cells. Using probes to chromosome‐specific sequences, it is possible to detect aneuploidy, and to document changes in specific chromosomes, which may have prognostic significance in some tumours, such as B‐cell chronic lymphatic leukaemia. Using sequence‐specific probes, translocations can be identified, such as the t(11;12) of Ewing's sarcoma. This review presents an outline of the technique of in situ hybridization and discusses areas of current and potential diagnostic application. © 1997 John Wiley & Sons, Ltd.

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Section: Preparation Of Tissuesmentioning

confidence: 99%

In Situ hybridization and its diagnostic applications in pathology

1997

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“…The reaction is strong in the smears that contain a large amount of microbial flora and is important in the detection of HSV from cervical samples, since numerous bacterial organisms are always contaminated. However, in the recent study by Kobayashi et a1.,30 the ISH technique using the same DNA probes applied to corneal herpetic lesions for the detection of HSV DNA in corneal scrapes showed nonspecific staining extremely seldom when compared to those of cervical specimens. These results suggested that the cytological ISH procedure might be applied to corneal herpetic lesions.…”

Section: Discussionmentioning

confidence: 95%

Comparison of immunocytochemistry and in situ hybridization in the cytodiagnosis of genital herpetic infection

Kobayashi¹

1992

Diagnostic Cytopathology

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Over a 62-month period, 53 patients were found to have cervicovaginal smears that contained cells consistent with, or equivocal cells for, a herpes simplex virus (HSV) infection. The Papanicolaou-destained smears from these cases were restrained in situ hybridization (ISH) with a biotinylated cloned DNA probe and immunocytochemistry (ICC) assay and were compared for the detection of HSV in cervicovaginal smears by two methods. Cytological findings classified the 53 slides into two groups, i.e., cytologically herpes positive (33 patients) and equivocal cases (20 patients). Each group was subdivided into two groups: group A was confirmed by ICC, and group B was confirmed by ISH technique. Of the 33 cellular samples containing cells considered to be consistent with a herpes infection, 15 (88%) of 17 were positive by means of ICC technique (group A), 6 (43%) of 14 were positive by ISH technique (group B). Of the 20 smears showing equivocal cell changes thought unlikely to be caused by an HSV infection, 6 (60%) of 10 were positive by ICC (group A), 2 (29%) of 7 were positive by ISH (group B). With the ISH technique, five smears showed dislodged cells from glass slides due to enzyme treatment and denaturation. The results revealed that the ICC technique is a rapid and reliable procedure and thus recommended for routine diagnosis of HSV infection. Moreover, ICC is easier to perform and interpret and is less expensive than ISH. Therefore, the ICC may be preferable to ISH for detecting HSV in routine Papanicolaou diagnostic work.

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“…On the other hand, cellular changes suggestive of herpes simplex virus (HSV) infection have been noted in different body sites, [3][4][5] including oral lesions. 6 Rapid methods of viral diagnosis in clinical samples include routine cytopathological examination for diagnostic viral inclusions and groundglass nuclei and immunocytochemical detection of specific viral antigens, 6 nucleic acid hybridization, 3,5 and most recently, a DNA diagnostic method using the polymerase chain reaction (PCR). 7,8 This newly described molecular biology technique is well-suited for the diagnosis of infection processes.…”

Section: Abstract: Pemphigus; Hsv Infection; Oral Cytology; Polymeramentioning

confidence: 99%

Coexistence of Pemphigus vulgaris and herpes simplex virus infection in oral mucosa diagnosed by cytology, immunohistochemistry, and polymerase chain reaction

Takahashi¹,

Kobayashi²,

Suzuki³

et al. 1998

Diagn. Cytopathol.

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The patient was a 53-yr-old woman, who presented in September 1994 with blisters on her oral mucosa, and later, numerous bullous lesions appeared on her lips, face, and head. The patient was admitted to the dermatological ward for further investigations. Laboratory examination was not remarkable except for immunoglobulin, i.e., IgA 581 mg/dl and IgG 2,504 mg/dl, while IgM levels were unremarkable. At about the same time, a dermatologist established the

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Cytological Diagnosis of Herpesvirus Keratitis by Means of in situ Hybridization: Report of a Case

Cited by 6 publications

References 7 publications

In Situ hybridization and its diagnostic applications in pathology

In Situ hybridization and its diagnostic applications in pathology

Comparison of immunocytochemistry and in situ hybridization in the cytodiagnosis of genital herpetic infection

Coexistence of Pemphigus vulgaris and herpes simplex virus infection in oral mucosa diagnosed by cytology, immunohistochemistry, and polymerase chain reaction

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