1562. Comparison of the Quidel AmpliVue HSV 1+2 Assay and Cepheid HSV Analyte Specific Reagents for Detection of Herpes Simplex Virus 1 and 2 in Clinical Specimens
Session: Poster Abstract Session: Clinical Infectious Diseases: Sexually Transmitted Infections
Saturday, October 10, 2015
Room: Poster Hall
Posters
  • AmpliVUE_HSV_poster_IDWeek_2015_092315.pdf (433.0 kB)
  • Background: We compare the FDA-cleared Quidel AmpliVue HSV 1+2 Assay (AmpliVue) to a laboratory developed test using Cepheid HSV-1 and -2 analyte specific reagents (LDT) for identification of HSV-1 and -2.

    Methods: Mucocutaneous swab specimens (n=231) were eluted into 3.0 ml viral transport medium (VTM) prior to AmpliVue and LDT testing.  For AmpliVue, a 20 µL portion of VTM was added to 1.6 mL of dilution buffer.  A 50 µL aliquot of the diluted sample was transferred to a reaction tube containing lyophilized reagents required for isothermal target amplification and labeling.  Following 45 min. incubation at 64C, reaction tubes were loaded into hand-held AmpliVue lateral flow devices for detection.  For LDT, 100 µL of VTM was heated at 95°C for 5 min.  and 5 µL of the specimen was combined with 15 µL of LDT master mix (1 bead each, HSV-1, -2 primer, probe, internal control; 5 SmartMix beads, 200 µL dH2O) for analysis by RT-PCR.  ELVIS cell culture was used as reference comparator.

    Results: Sensitivity and specificity of AmpliVue and LDT for identification of HSV-1 and -2 compared to ELVIS are presented below.  Analysis of discrepant results between either molecular assay and ELVIS was based on a third PCR-based assay and DNA sequence analysis to confirm HSV type.  This resolution supported the presence of HSV-1 in 6/9 (AmpliVue) and 3/7 (LDT) specimens categorized as false positive based on ELVIS result.  This included 4 specimens that were positive by AmpliVue but negative by ELVIS and LDT.  Similarly, the presence of HSV-2 was confirmed in 11/16 (AmpliVue) and 11/21 (LDT) specimens categorized as false positive. This suggests the presence of low titer or non-viable HSV in these specimens.  Post-resolution sensitivity and specificity was as follows: AmpliVue sensitivity; 97.7% (HSV-1), 96.4% (HSV-2), specificity; 98.4% (HSV-1), 97.1% (HSV-2), LDT sensitivity; 84.6% (HSV-1), 100% (HSV-2), specificity; 97.9% (HSV-1), 94.3% (HSV-2).

     

    TP

    FP

    TN

    FN

    Total

    Sensitivity (CI)

    Specificity (CI)

    AmpliVue

     

     

     

     

     

     

     

         HSV-1

    37

    9

    183

    1

    230

    97.4% (85-99)

    95.3% (91-98)

         HSV-2

    42

    16

    169

    3

    230

    93.3% (81-98)

    91.4% (86-95)

    LDT

     

     

     

     

     

     

     

         HSV-1

    36

    7

    186

    2

    231

    94.7% (81-99)

    96.4% (92-98)

         HSV-2

    44

    21

    165

    1

    231

    97.8% (87-99)

    88.7% (83-93)

    Conclusion: AmpliVue HSV 1+2 and Cepheid HSV-1 and -2 analyte specific reagents (LDT) demonstrated comparable sensitivity and specificity for the detection of HSV-1 and -2.  Both tests identified HSV in specimens that were negative by ELVIS culture.

    Blake Buchan, PhD, D(ABMM), Pathology, Medical College of Wisconsin, Milwaukee, WI and Nathan Ledeboer, PhD, D(ABMM), Microbiology, Medical College of Wisconsin, Milwaukee, WI

    Disclosures:

    B. Buchan, Quidel: Research Contractor , Research support

    N. Ledeboer, None

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