Detection of herpes simplex virus infection of female genitalia by the peroxidase-antiperoxidase method alone or in conjunction with the Papanicolaou stain

Abstract

The sensitive peroxidase-antiperoxidase (PXAPX) method individually and in conjunction with the Papanicolaou (PAP) stain was used to detect herpes simplex virus (HSV) in specimens from human female genitalia. Initial studies using a model system of HSV-1or HSV-2-infected Vero cells established (i) acetone as the most effective fixative, (ii) optimal dilutions of preimmunization and anti-HSV serum for differentiation of infected from noninfected cells, (iii) optimal concentration of 3,3'-diaminobenzidine tetrahydrochloride and H202 for maximal staining of infected cells with minimal background reaction, and (iv) removal of endogenous peroxidase with absolute MeOH. These various parameters, once established, were utilized in the PXAPX or PXAPX-PAP on human specimens from the vulva or cervix. In these specimens, examined by standard light microscopy, PXAPX-positive cells were dark brown with a single nucleus or multiple nuclei. By coupling the PAP to the PXAPX, detailed nuclear observations of PXAPX-positive cells were possible and revealed nuclear changes consistent with HSV infection, including syncytium formation, chromatin condensation, and an occasional Cowdry type A inclusion. The PXAPX and PXAPX-PAP correlated (r = 0.742) over a period of 72 h with HSV isolation from these samples. MATERIALS AND METHODS Virus. Virus stocks of HSV-1 (strain 69-85) and HSV-2 (strain 316-D) were obtained from F.

Comparison of direct immunofluorescence and direct immunoperoxidase procedures for detection of herpes simplex virus antigen in lesion specimens

“…Although DIF and DIP showed good agreement with virus isolation, the sensitivity of these methods for the detection of HSV-positive specimens was somewhat lower than that seen in certain other studies (1,3,9,10,13,16). This may be related to two factors.…”

“…In our accumulated experience and in reports of various other workers, IF methods have shown well over 80% agreement with cell culture isolation for detection of HSV in clinical materials (9,12,13,16), although some studies (2) have shown a lower sensitivity for IF. Immunoperoxidase (IP) staining procedures also have been advanced in recent years for detection of HSV in lesion materials (1,3,8,9,10). These are considered to have advantages over IF methods in that results can be read with an ordinary light microscope and permanent preparations can be made.…”

Use of the Immunoperoxidase Technique for Estimation of Susceptibility of Herpes Simplex Virus to Idu

“…The immunoperoxidase antibody method has been employed in the last few years in clinical virology for virus identification in the place of fluorescent antibody technique (Benjamin 1977;Person et a. 1979).…”

Flow cytometric detection of herpes simplex virus type 2 infected and transformed cells by immunoenzymatic and by indirect immunofluorescence staining

“…Immunoperoxidase staining has been the It is possible to detect cells stained with an absorbing method of choice using either the indirect conjugated dye by light scatter analysis in flow cytometry systems antibody method (3,4,14) or the unlabeled antibody-en- (9)(10)(11)14). We have shown by calculation and experiment zyme method (9,16,17). The effectiveness of using the that the decrement in light scatter intensity caused by immunoperoxidase technique on human specimens, the addition of a n absorbing dye can be enhanced by however, is compromised by the endogenous peroxidase restricting the collection of scattered light to the 3"-6" activity commonly found in mammalian mucosecretions angular interval (9).…”