Clear detection and typing of herpes simplex virus types 1 and 2 by an indirect ELISA assay: Comparison with three different combined methods—capture ELISA, restriction enzymes, and polymerase chain reaction

Markoulatos, Panayotis; Fountoucidou, Polyxeni; Marinakis, George; Krikelis, V.; Spyrou, Niki; Vamvakopoulos, Nicholas C.; Moncany, Maurice L. J.

doi:10.1002/(sici)1098-2825(1997)11:3<146::aid-jcla5>3.0.co;2-8

Cited by 11 publications

(8 citation statements)

References 30 publications

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“…No isolate needed to be further typed by another method. Our data also were reinforced by the fact that 6 HSV-1 and 14 HSV-2 isolates that had been included in a previous study (8) and were analyzed by 4 different methods; indirect ELISA, capture ELISA, restriction enzymes DNA analysis, and PCR; were tested by the present multiplex PCR assay. All these 20 isolates were detected and typed in perfect concordance with the previous reported results.…”

Section: Detection and Typing Of Clinical Isolates By Multiplex Pcrsupporting

confidence: 56%

“…Analysis of viral DNA with restriction enzymes is a specific method and the restriction patterns of HSV-1 are readily distinguished from those of HSV-2. Typing of HSV isolates by restriction-enzyme-DNA analysis is time consum-ing, involves the use of radioisotopes, and is too cumbersome for most clinical laboratories to be performed on a large scale (7,8).…”

Section: Introductionmentioning

confidence: 99%

“…Sensitivity and specificity may vary with different primer pairs and the genetic variability of naturally occurring strains of viruses is an important influencing factor. Many different primer pairs have been suggested for the diagnosis of HSV and VZV infections (1,2,8,(11)(12)(13)(14)(15).…”

Section: Introductionmentioning

confidence: 99%

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Detection and typing of HSV-1, HSV-2, and VZV by a multiplex polymerase chain reaction

Markoulatos

Georgopoulou

Kotsovassilis

et al. 2000

J. Clin. Lab. Anal.

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The development of a multiplex polymerase chain reaction method for the rapid and accurate detection and typing of HSV‐1, HSV‐2, and VZV from clinical specimens is described. A sensitive multiplex polymerase chain reaction was achieved by optimization of parameters such as the primers, magnesium, and dNTPs concentrations. False‐negative results that sometimes arise due to inhibitors of DNA amplification or failure of DNA extraction procedure used may be avoided by assaying each specimen with α‐tubulin primers. Multiplex PCR amplified viral sequences from all 55 specimens obtained from patients with clinical evidence of HSV or VZV infection indicated 100% sensitivity. From 55 patients who were investigated by multiplex PCR, HSV‐1 was detected in 28, HSV‐2 in 20, and VZV in 7 specimens. The reported results indicate that the present multiplex PCR assay has a potential application in clinical diagnosis when a rapid and accurate detection and typing of involved viruses HSV‐1, HSV‐2, or VZV is needed. J. Clin. Lab. Anal. 14:214–219, 2000. © 2000 Wiley‐Liss, Inc.

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Section: Detection and Typing Of Clinical Isolates By Multiplex Pcrsupporting

confidence: 56%

Section: Introductionmentioning

confidence: 99%

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Detection and typing of HSV-1, HSV-2, and VZV by a multiplex polymerase chain reaction

Markoulatos

Georgopoulou

Kotsovassilis

et al. 2000

J. Clin. Lab. Anal.

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“…Improvements in antibody production in recent years providing monospecific polyclonal and monoclonal antibodies has increased the efficiency of typing (Coyle et al, 1999; Markoulatos et al, 1997). Specifically, antibodies raised against type specific envelope glycoproteins are allowing for a much more accurate diagnosis, providing better sensitivity and specificity (Liljeqvist, Svennerholm, and Bergstrom, 1999; Slomka et al, 1998).…”

Section: Discussionmentioning

confidence: 99%

“…In the case of HSV-1, the severity can range from simple cold sores to fatal encephalitis (Markoulatos et al, 1997). Vertical transmission of HSV-2 can result in disseminated neonatal infections which can have devastating consequences (Corey, 1994; Coyle et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

Reconsideration of viral protein immunoblotting for differentiation of human herpes simplex viruses

Bowles¹,

Yedowitz²,

Blaho³

2008

Diagnostic Microbiology and Infectious Disease

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Herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) are ubiquitous human pathogens which infect their hosts for life and reactivate to cause disease at or near the initial site of infection. As the incidence of genital HSV-1 infections increase, there is an increased demand for valid viral typing diagnostics. In this report, we reconsidered and developed a triple-phase, immune-typing procedure that compares differences in electrophoretic mobilities of viral ICP4, ICP27, and VP22 proteins between HSV-1 and HSV-2 strains. We isolated and immunotyped five primary HSV-1 strains derived from oro-facial, ocular, and genital areas along with two primary HSV-2 strains from the genital area. Advantages of this methodology include its general technical simplicity, sensitivity, and ability to definitively type HSV. It is anticipated that this methodology will be useful in distinguishing viruses obtained in clinical cultures.

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Development of a quadriplex polymerase chain reaction for human cytomegalovirus detection

Markoulatos

Samara

Siafakas

et al. 1999

J. Clin. Lab. Anal.

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Clear detection and typing of herpes simplex virus types 1 and 2 by an indirect ELISA assay: Comparison with three different combined methods—capture ELISA, restriction enzymes, and polymerase chain reaction

Cited by 11 publications

References 30 publications

Detection and typing of HSV-1, HSV-2, and VZV by a multiplex polymerase chain reaction

Detection and typing of HSV-1, HSV-2, and VZV by a multiplex polymerase chain reaction

Reconsideration of viral protein immunoblotting for differentiation of human herpes simplex viruses

Development of a quadriplex polymerase chain reaction for human cytomegalovirus detection

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