1887

Abstract

. Herpes simplex virus (HSV) is a common lifelong sexually transmitted infection. HSV-1 typically manifests as oral cold sores, while HSV-2 is more traditionally associated with sexual transmission and infection. We have developed a real-time PCR (Trioplex) for the simultaneous detection of HSV-1 and -2 and the bacterial phage internal control (IC) MS2.

. To determine the performance of the Trioplex method and resolve discrepancies, 178 clinical specimens from cutaneous and mucocutaneous sources were tested using 3 different methods; virus culture with direct fluorescent antibody (DFA) immunostaining, Trioplex and a commercially available HSV analyte-specific reagent (ASR).

. HSV Trioplex was significantly more sensitive than virus culture (89 and 67 % HSV 1/2, respectively) and comparable to the commercial assay (<0.001). Cost analysis revealed that the Trioplex reduced cost by 80  % compared to cell culture.

. The implementation of the HSV Trioplex improved the detection turnaround time from 3–10 days to 2.5 h, thus streamlining Herpes detection, improving sensitivity and reducing laboratory costs.

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/content/journal/jmm/10.1099/jmm.0.000971
2019-05-01
2019-10-18
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References

  1. Liu J, Yi Y, Chen W, Si S, Yin M et al. Development and evaluation of the quantitative real-time PCR assay in detection and typing of herpes simplex virus in swab specimens from patients with genital herpes. Int J Clin Exp Med 2015;8:18758–18764
    [Google Scholar]
  2. Looker KJ, Magaret AS, May MT, Turner KME, Vickerman P et al. Global and regional estimates of prevalent and incident herpes simplex virus type 1 infections in 2012. PloS One 2015;10:e0140765
    [Google Scholar]
  3. Looker KJ, Magaret AS, Turner KME, Vickerman P, Gottlieb SL et al. Global estimates of prevalent and incident herpes simplex virus type 2 infections in 2012. PloS One 2015;10:e114989
    [Google Scholar]
  4. LeGoff J, Péré H, Bélec L. Diagnosis of genital herpes simplex virus infection in the clinical laboratory. Virol J 2014;11:83 [CrossRef]
    [Google Scholar]
  5. Gupta R, Warren T, Wald A. Genital herpes. The Lancet 2007;370:2127–2137 [CrossRef]
    [Google Scholar]
  6. Lafferty WE, Krofft S, Remington M, Giddings R, Winter C et al. Diagnosis of herpes simplex virus by direct immunofluorescence and viral isolation from samples of external genital lesions in a high-prevalence population. J Clin Microbiol 1987;25:323–326
    [Google Scholar]
  7. Curtin WM, Menegus MA, Patru MM, Peterson CJ, Metlay LA et al. Midtrimester fetal herpes simplex-2 diagnosis by serology, culture and quantitative polymerase chain reaction. Fetal Diagn Ther 2013;33:133–136
    [Google Scholar]
  8. Aliabadi N, Jamalidoust M, Asaei S, Namayandeh M, Ziyaeyan M. Diagnosing of herpes simplex virus infections in suspected patients using real-time PCR. Jundishapur J Microbiol 2015;8:e16727 [CrossRef]
    [Google Scholar]
  9. Tan TY, Zou H, Ong DCT, Ker KJ, Chio MTW et al. Development and clinical validation of a multiplex real-time PCR assay for herpes simplex and varicella zoster virus. Diagn Mol Pathol 2013;22:245–248 [CrossRef]
    [Google Scholar]
  10. Corey L, Huang ML, Selke S, Wald A. Differentiation of herpes simplex virus types 1 and 2 in clinical samples by a real-time TaqMan PCR assay. J Med Virol 2005;76:350–355 [CrossRef]
    [Google Scholar]
  11. Kimberlin DW. The tail wagging the dog (or the challenges faced when the financing of medicine gets ahead of the science of Medicine). J Clin Microbiol 2018;56:e00904–00918 [CrossRef]
    [Google Scholar]
  12. Buelow DR, Bankowski MJ, Fofana D, Gu Z, Pounds S et al. Comparison of two multiplexed PCR assays for the detection of HSV-1, HSV-2, and VZV with extracted and unextracted cutaneous and mucosal specimens. J Clin Virol 2013;58:84–88 [CrossRef]
    [Google Scholar]
  13. Young S, Body B, Moore F, Dunbar S. Multicenter evaluation of the Luminex® ARIES® HSV 1&2 Assay for the detection of herpes simplex virus types 1 and 2 in cutaneous and mucocutaneous lesion specimens. Expert Rev Mol Diagn 2016;16:1241–1249 [CrossRef]
    [Google Scholar]
  14. Lai W, Chen CY, Morse SA, Htun Y, Fehler HG et al. Increasing relative prevalence of HSV-2 infection among men with genital ulcers from a mining community in South Africa. Sex Transm Infect 2003;79:202–207 [CrossRef]
    [Google Scholar]
  15. Rolfe KJ, Parmar S, Mururi D, Wreghitt TG, Jalal H et al. An internally controlled, one-step, real-time RT-PCR assay for norovirus detection and genogrouping. J Clin Virol 2007;39:318–321 [CrossRef]
    [Google Scholar]
  16. Dreier J, Störmer M, Kleesiek K. Use of bacteriophage MS2 as an internal control in viral reverse transcription-PCR assays. J Clin Microbiol 2005;43:4551–4557 [CrossRef]
    [Google Scholar]
  17. Hoorfar J, Malorny B, Abdulmawjood A, Cook N, Wagner M et al. Practical considerations in design of internal amplification controls for diagnostic PCR assays. J Clin Microbiol 2004;42:1863–1868 [CrossRef]
    [Google Scholar]
  18. Sedmak G, Bina D, MacDonald J, Couillard L. Nine-year study of the occurrence of culturable viruses in source water for two drinking water treatment plants and the influent and effluent of a wastewater treatment plant in Milwaukee, Wisconsin (August 1994 through July 2003). Appl Environ Microbiol 2005;71:1042–1050 [CrossRef]
    [Google Scholar]
  19. Jeddi F, Piarroux R, Mary C. Application of the NucliSENS easyMAG system for nucleic acid extraction: optimization of DNA extraction for molecular diagnosis of parasitic and fungal diseases. Parasite 2013;20:52 [CrossRef]
    [Google Scholar]
  20. Viera AJ, Garrett JM. Understanding interobserver agreement: the kappa statistic. Fam Med 2005;37:360–363
    [Google Scholar]
  21. Koelle DM, Wald A. Herpes simplex virus: the importance of asymptomatic shedding. J Antimicrob Chemother 2000;45:1–8 [CrossRef]
    [Google Scholar]
  22. Faron ML, Ledeboer NA, Patel A, Beqa SH, Yen-Lieberman B et al. Multicenter evaluation of the meridian bioscience molecular HSV 1&2 assay for the detection of herpes simplex virus 1 and 2 from clinical cutaneous and Mucocutaneous specimens. J Clin Microbiol 2016;54:2008–2013 [CrossRef]
    [Google Scholar]
  23. Fan F, Stiles J, Mikhlina A, Lu X, Babady NE et al. Clinical validation of the Lyra direct HSV 1+2/VZV assay for simultaneous detection and differentiation of three herpesviruses in cutaneous and mucocutaneous lesions. Loeffelholz MJ, editor J Clin Microbiol 2014;52:3799–3801 [CrossRef]
    [Google Scholar]
  24. Patwardhan V, Bhalla P, Rawat D, Garg VK, Sardana K et al. A comparative analysis of polymerase chain reaction and direct fluorescent antibody test for diagnosis of genital herpes. J Lab Physicians 2017;9:53–56
    [Google Scholar]
  25. Rübben A, Baron JM, Grussendorf-Conen EI. Routine detection of herpes simplex virus and varicella zoster virus by polymerase chain reaction reveals that initial herpes zoster is frequently misdiagnosed as herpes simplex. Br J Dermatol 1997;137:259–261 [CrossRef]
    [Google Scholar]
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