2. Preliminary evaluation of the analytical and
clinical performances multiplex RT-PCR DNA
microarray,
the Clart Enthepex kit.

3. This kit has been developed to determine in a fast
way the agent(s) responsible for the illness
(Herpesvirus or Enterovirus), thus allowing the
establishment of the most effective clinical
treatment in each case.

4. This kit is based on viral genome-specific fragment
amplification
by multiplex PCR and its
subsequent detection via
hybridization with a
microorganism-specific
binding probe on
low-density microarrays.

5.  allows simultaneous detection and identification of
the eight human herpesviruses and enteroviruses.
Coxsackivirus
 HHV-8

6.  HSV-Herpes Simplex virus
 VZV-Varicella Zoster virus
 CMV- Cytomegalovirus
 EBV-Epstein-Barr virus
 HHV-6, HHV-7, HHV-8 -human herpesvirus
 HEVs – human enteroviruses

8. Since neither QCMD panels nor known clinical
samples containing HHV-7 or HHV-8 were available,
the performances of the DNA microarray regarding
the detection of both these viruses have not been
evaluated.

9. 4 proficiency samples for each of HSV-1, HSV-2,
VZV, CMV, EBV, HHV-6 and HEVs with
concentrations ranging from 10² to 10⁵ copies/ml
were selected and independently tested 4 times
each in order to assess the specificity, the
sensitivity, and reproducibility of the DNA
microarray.

10.  Limit of detection of less than 500 copies/ml for all
of the 6 herpesviruses tested.
 HEVs- the low viral loads corresponding to 280
and 390 genome copies/ml were not detected
 HEVs – the viral load corresponding to
1480 genome copies/ml was detected in 3 out 4
analyses (75%)

11. The analytical sensitivities of the test corresponding
to the copy number of each one of the viruses that
can be detected in 100% of the analyses performed
were :
 HHV-6 between 500 to 1,000 copies/ml
 HSV-1, HSV-2, VZV, and EBV >2,000 copies/ml

13. The manufacturer's claims regarding the analytical
sensitivities of the assay are:
 10 copies for VZV, HHV-7, and HSV-1
 100 copies for HSV-2, CMV, EBV, HHV-6, HHV-8, and
the coxsackieviruses
 1,000 copies for the echoviruses and the polioviruses
per PCR mixture containing 5 μl of DNA/RNA extract.

14. Retrospective analysis of 78 CSF samples obtained
from patients hospitalized for suspected
neurological virus infections from 2002 to 2009

15.  28 samples previously tested positive for
herpesviruses (7 HSV-1, 2 HSV-2, 8 VZV, 1 CMV,
4 EBV, 6 HHV-6) and negative for HEV
 30 samples previously positively tested for HEV
and negative for herpesviruses.
 20 samples previously tested negative for
both HEVs and herpesviruses.

16.  27 of the 28 herpesvirus-positive CSF samples
were detected (96%)
 30 of the 30 HEV-positive CSF samples were
detected (100%)
 Were detected 11 (37%) HHV-7 and HEV mixed
infections among the 30 pediatric aseptic
meningitis cases initially related to HEVs

20.  High sensitivity allowing the detection of
minimum quantities of DNA.
 High specificity, when using a sequence
corresponding to a highly preserved region within
the viral genome and binding probes specific to
each human herpes and enterovirus type.
 Fast. Allowing the laboratory to provide the
answer to the clinician in a single work day
 Simultaneous detection of multiple DNA & RNA
viruses present in a single sample

21.  Limited automation (requires numerous handlings)
 Kit cannot be used without a microarray reader
piloted by specific software provided by the
manufactures, thus limiting the implementation of
the kit
 Risk of contamination during handling amplicons

22. Due to the small number of CSF specimens tested,
further studies are needed to better characterize the
performances of this test before its use in routine
patient care.

24.  http://www.genomica.es/en/in_vitro_diagnostics_products_clart_entherpex.cfm
 http://www.qcmd.org/index.php?pageId=34&pageVersion=EN
 http://www.journalofclinicalvirology.com/article/S1386-6532(02)00028-8
 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3522074
 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3209095/?log$=activity