Evaluation of a transcription mediated amplification assay for detection of herpes simplex virus types 1 and 2 mRNA in clinical specimens

• Published in: Journal of clinical virology Vol. 80; pp. 62 - 67
• Main Authors: Swenson, Paul D., El-Sabaeny, Azza, Thomas-Moricz, Vanessa, Allen, Megan, Groskopf, Anabel, Jiang, Alice, Getman, Damon
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.07.2016
• Subjects: Allergy and Immunology; Aptima; Cytomegalovirus; Female; Herpes Simplex - diagnosis; Herpes simplex virus (HSV); Herpes simplex virus 1; Herpes simplex virus 2; Herpesvirus 1, Human - genetics; Herpesvirus 1, Human - growth & development; Herpesvirus 2, Human - genetics; Herpesvirus 2, Human - growth & development; Human herpesvirus 3; Human herpesvirus 4; Human herpesvirus 5; Human herpesvirus 6; Human herpesvirus 8; Humans; Infectious Disease; Male; Real-Time Polymerase Chain Reaction; RNA, Messenger - analysis; RNA, Viral - analysis; Sensitivity and Specificity; Transcription, Genetic; Transcription-mediated amplification; UL42 mRNA; Aptima; UL42 mRNA; Herpes simplex virus (HSV); Transcription-mediated amplification
• ISSN: 1386-6532; 1873-5967; 1873-5967
• DOI: 10.1016/j.jcv.2016.04.023

•Herpes simplex viruses (HSVs) are double-stranded DNA viruses that package viral mRNAs.•A real time amplified molecular test was designed to detect packaged viral mRNA.•This test exhibited high clinical sensitivity and specificity in an STD clinic population. Herpes simplex viruses (HSV) are double-stranded DNA human herpesviruses (HHVs) that have the capacity to cause significant morbidity and mortality in humans. Like HHV5 (Cytomegalovirus) and HHV8 (Kaposi’s sarcoma virus), HSV type 1 (HSV-1), and HSV type 2 (HSV-2) (HHV1, HHV2) selectively package certain viral messenger RNAs inside mature virions, as well as expressing those mRNAs in infected cells. To evaluate the clinical and analytical performance of Aptima HSV 1&2 assay (AHSV), a newly developed automated real time transcription-mediated amplification (TMA) nucleic acid amplification test (NAAT) for HSV-1 and 2 UL42 mRNAs, compared to viral culture and HSV DNA NAAT. Cutaneous and mucocutaneous lesion swab specimens from a population of symptomatic female and male subjects attending a U.S. public health clinic (n=758) were evaluated by shell vial culture with fluorescent antibody staining for HSV-1 and 2. Specimens were then tested with AHSV for HSV-1 and 2 on the Panther instrument. Specimens from subjects with discordant culture—TMA paired results were tested using an FDA-cleared test for HSV-1 and 2 viral DNA. Analytical performance of AHSV was evaluated using test panels consisting of laboratory strains of HSV-1 and 2 and a variety of non-target human DNA viruses. Compared to culture, AHSV was sensitive and specific for detection of HSV-1 and 2 in patient lesion swab specimens, exhibiting clinical sensitivities of 98.2% (95% CI: 92.9–99.7) and 99.4% (95% CI: 96.0–99.9), respectively. Addition of HSV DNA NAAT discordant resolution testing results to culture results improved AHSV sensitivity for HSV-1 and 2–99.2% (95% CI: 94.7–99.9) and 100% (95% CI: 97.5–100), respectively. Clinical specificity of AHSV for HSV-1 and 2 detection was 97.8% (95% CI: 96.3–98.8) and 94.5% (95% CI: 92.2–96.1), respectively, compared to culture; and 99.5% (95% CI: 98.5–99.9) and 99.5% (95% CI: 98.3–99.7), respectively, compared to culture with discordant resolution. Analytical sensitivity (95% limit of detection) of AHSV for HSV-1 (McIntyre strain) was 28.9 TCID50/mL (95% FL: 23.4–37.9), and 0.54 TCID50/mL (95% FL: 0.42–0.75) for HSV-2 (MS strain). AHSV did not cross-react with laboratory strains of HHV-3, HHV-4, HHV-5, HHV-6, and four other non-HHV human DNA viruses. Real time transcription-mediated amplification NAAT for HSV viral mRNA is a sensitive and specific method for detection of herpes simplex virus infection in symptomatic patients.