24th Clinical Virology Symposium
April 27 - 30, 2008 Daytona Beach, Florida, USA
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Conference Sessions
Session I
Session II
Session III
Session IV
 

REAL-TIME PCR DETECTION OF HERPES SIMPLEX VIRUS IN CSF USING THE SPARTAN DXTM

Session ID: S32
Author Name: L. Shaw1, A. Rogaeva2, C. Harder2, and T. Karnauchow1,3 1Regional Virology Laboratory, Children?s Hospital of Eastern Ontario; 2 Spartan Bioscience Inc.; and 3University of Ottawa, Ottawa, ON, Canada
Country: Canada
Conference Session: Session I

 

The Spartan DXTM (Spartan Bioscience Inc., Ottawa ON) is a compact, inexpensive, 4-well real-time PCR (RTPCR) instrument. It uses a 2-temperature reaction cycle that promises more rapid results than conventional RTPCR.  The diagnosis of HSV infections of the CNS is a logical application for this instrument: on-demand, non-batched analysis where speed and convenience are critical.  We evaluate the performance of a TaqMan®-based HSV assay adapted for the Spartan DX.

Methods: The RTPCR HSV assay used at CHEO (“reference-HSV”) was modified for the Spartan DXTM (“Spartan-HSV”).  Replicate testing of CSF spiked with HSV-1 DNA and with HSV-2 DNA (180 copies and 160 copies, respectively; ABi Inc., Columbia, MD) was performed.  HSV-1 stock (3.2 TCID50; 1 log > the lower limit of detection of reference-HSV) was added to blank CSF, extracted, and tested in same- and separate-day runs.  Finally, archived CSF specimens (23 PCR[+], 10 PCR[-]) were tested.  Extractions were performed by QIAamp DNA Mini Kit.  Specimens were tested in parallel by reference-HSV (ABI 7500; BioRad iCycler).  Spartan- and reference-HSV results (PCR [+/-], CT) were compared.

Results: Spartan-HSV detected both HSV-1 and HSV-2 DNA: mean CT values of samples spiked with 180 copies HSV-1/reaction (n=8) and with 160 copies HSV-2/reaction (n=8), were 35 (range: 34-36).  For CSF samples spiked with 3.2 TCID50 HSV-1 (n=46), the mean CT was 36.6 (range: 35-39).  For all study samples, Spartan and reference test results were in agreement. For clinical specimens, there was no meaningful CT difference between methods (Spartan CT = reference ± 2 CT). 

Conclusions: The Spartan-HSV assay is reliable and reproducible, with performance equaling conventional RT HSV PCR.  The Spartan DXTM is an excellent choice for laboratories needing an accurate, affordable, convenient platform to accommodate non-batched “stat” testing, or for smaller laboratories seeking to implement real-time molecular diagnostics in their facilities.