Detection of Herpesviridae in whole blood by multiplex PCR DNA-based microarray analysis after hematopoietic stem cell transplantation.
Viral infections are important causes of morbidity and mortality in patients after hematopoietic stem cell transplantation. The monitoring by PCR of Herpesviridae loads in blood samples has become a critical part of posttransplant follow-up, representing mounting costs for the laboratory. In this study, we assessed the clinical performance of the multiplex PCR DNA microarray Clart Entherpex kit for detection of cytomegalovirus (CMV), Epstein-Barr virus (EBV), and human herpesvirus 6 (HHV-6) as a screening test for virological follow-up. Two hundred fifty-five blood samples from 16 transplanted patients, prospectively tested by routine PCR assays, were analyzed by microarray. Routine PCR detected single or multiple viruses in 42% and 10% of the samples, respectively. Microarray detected single or multiple viruses in 34% and 18% of the samples, respectively. Microarray results correlated well with CMV and EBV detections by routine PCR (kappa tests = 0.79 and 0.78, respectively), whereas a weak correlation was observed with HHV-6 (0.43). HHV-7 was also detected in 48 samples by microarray. In conclusion, the microarray is a reliable screening assay for a posttransplant virological follow-up to detect CMV and EBV infections in blood. However, positive samples must be subsequently confirmed and viral loads must be quantified by PCR assays. Limitations were identified regarding HHV-6 detection. Although it is promising, is easy to use as a first-line test, and allows a reduction in the cost of analysis without undue delay in the reporting of the final quantitative result to the clinician, some characteristics of this microarray should be improved, particularly regarding quality control and the targeted virus panel, such that it could then be used as a routine test.
Center for Molecular Diagnostic, iris-Lab, Iris-Brussels Public Hospital Network, Brussels, Belgium fdebaugn@ulb.ac.be.
France
F
Debaugnies
Department of Microbiology, iris-Lab, Iris-Brussels Public Hospital Network, Brussels, Belgium.
Laurent
L
Busson
Department of Haematology and Oncology, Hôpital Universitaire Des Enfants Reine Fabiola, Brussels, Belgium.
Alina
A
Ferster
Department of Haematology, Jules Bordet Institute, Brussels, Belgium.
Philippe
P
Lewalle
Department of Haematology and Oncology, Hôpital Universitaire Des Enfants Reine Fabiola, Brussels, Belgium.
Nadira
N
Azzi
Division of Infectious Diseases, Jules Bordet Institute, Brussels, Belgium.
Mickael
M
Aoun
Center for Molecular Diagnostic, iris-Lab, Iris-Brussels Public Hospital Network, Brussels, Belgium.
Godelieve
G
Verhaegen
Department of Microbiology, iris-Lab, Iris-Brussels Public Hospital Network, Brussels, Belgium.
Bhavna
B
Mahadeb
Center for Molecular Diagnostic, iris-Lab, Iris-Brussels Public Hospital Network, Brussels, Belgium.
Jérôme
J
de Marchin
Department of Microbiology, iris-Lab, Iris-Brussels Public Hospital Network, Brussels, Belgium.
Olivier
O
Vandenberg
Center for Molecular Diagnostic, iris-Lab, Iris-Brussels Public Hospital Network, Brussels, Belgium.
Marie
M
Hallin
D000293:Adolescent; D000328:Adult; D001769:Blood / Q000821:virology*; D002648:Child; D002675:Child, Preschool; D003587:Cytomegalovirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects; D006566:Herpesviridae Infections / Q000175:diagnosis* / Q000821:virology; D004854:Herpesvirus 4, Human / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D015654:Herpesvirus 6, Human / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016867:Immunocompromised Host; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D008403:Mass Screening / Q000379:methods; D046228:Microarray Analysis / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000379:methods; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*