Evaluation of three immunofluorescence assays for culture confirmation and typing of herpes simplex virus

Abstract

(KL), were evaluated for the laboratory confirmation and typing of herpes simplex virus (HSV). Of 108 coded HSV slide preparations, run in parallel with each monoclonal-antibody set, 103 were equivalent by the immunofluorescence assays. Among the five discordant isolates, three (2.8%) did not type with the KL monoclonal antibodies and two (1.9%) false-positive results occurred with the Syva typing system. All of the HSV clinical isolates tested were correctly typed with the ENI indirect immunofluorescence antigen detection system. Typing confirmation of the five discordant HSV isolates was performed by differential sensitivity to 5-bromo-2'-deoxyuridine and endonuclease cleavage analysis of the viral DNA. Use of the Syva and KL direct immunofluorescence antigen detection systems for the identification of HSV isolates is less time-consuming than use of the ENI indirect antigen detection system; however, sensitivity and specificity may be lost. * Corresponding author. medium (Hanks balanced salt solution) containing 0.5% gelatin, 50 ,ug of gentamicin per ml, and 100 U of mycostatin per ml. Specimens not immediately sent to the Virology Laboratory were temporarily stored (i.e., 24 to 38 h) at 4°C. Specimens were received from Nassau County Medical Center clinics, more than 13 hospitals, and approximately 30 physicians in group or individual practice throughout Nassau, Suffolk, Queens, and Manhattan Counties, N.Y. Virus isolation. Ail specimens received by the Virology

Evaluation of five monoclonal antibody-based kits or reagents for the identification and culture confirmation of herpes simplex virus

“…The Syva typing kit resulted in three false-positive HSV-2 signals, giving the erroneous indication of dual HSV-1 and HSV-2 infections. Previous studies in our laboratory with the Syva kit incorrectly identified 2 of 108 positive specimens as dual infections (12). In reports of recent studies, Gleaves and coworkers (9, 10) reported a total of three dual isolates obtained with the Syva kit.…”

“…Collection and transport of * Corresponding author. specimens to the laboratory have been described previously (12).…”

“…Additionally, the intensities and numbers of immunofluorescent cells in individually infected cultures were determined and found to be approximately equal prior to mixing. The plaque and immunofluorescence staining assays were performed by routine procedures (12,17). Analysis of the HSV DNA in coculture and individually infected cultures was performed as described above.…”

Typing of Clinical Herpes Simplex Virus Type 1 and Type 2 Isolates with Monoclonal Antibodies

“…Since several HSV-specific monoclonal antibodies (MAbs) have been shown to identify type-specific epitopes (2,5,11,17), these reagents have been used for type-specific antigen detection. However, the reported sensitivities and specificities for such reagents, including commercially available kits, have been found to vary among the assays in the range between 92 and 100% (4,6,7,17). This discrepancy may be explained by variability in the expression of different HSV proteins or intratypic variability of different epitopes within the same protein among the isolates (11).…”

Detection of cutaneous herpes simplex virus infections by immunofluorescence vs. PCR

“…Direct identification of the virus can be done with cell culture, which is rather timeconsuming, electron microscopy, or antigen detection, quite often based on immunofluorescence techniques. [1][2][3][4][5][6][7] Reported sensitivities of different techniques vary widely. In addition, amplification of DNA from skin swabs by polymerase chain reaction (PCR) has also been established.…”