Improved multiplex-PCR and microarray for herpesvirus detection from CSF.
BACKGROUND
A multiplex-PCR and microarray-based method was designed for detection of eight herpesviruses from clinical specimens.


OBJECTIVE
To improve the method, especially for detection of herpes simplex (HSV) and varicella-zoster viruses (VZV), and to update and validate the method using positive cerebrospinal fluid (CSF) samples.


METHODS
A new primer pair for HSV-PCR and four detection oligonucleotides for HSVs were designed. Two new detection oligonucleotides for VZV and additional oligonucleotides for CMV, EBV, HHV-6 and -7 were designed. The new and previous multiplex-PCR and microarray method were tested in parallel with dilution series of commercial herpesvirus DNAs and 20 CSF specimens positive for HSV-1, HSV-2, or VZV.


RESULTS
New method was more sensitive for detection of HSVs and both two new detection oligonucleotides for VZV functioned well at low levels of viral DNA.


CONCLUSIONS
The revised HSV-PCR and new HSV- and VZV-oligonucleotides were found to function well and be more sensitive, thereby increasing reliability of the method.
Jääskeläinen
Anne J
AJ
Department of Virology, Haartman Institute, FIN-00014 University of Helsinki, Finland. anne.jaaskelainen@helsinki.fi
Piiparinen
Heli
H
Lappalainen
Maija
M
Vaheri
Antti
A
D020805:Central Nervous System Viral Diseases / Q000134:cerebrospinal fluid / Q000175:diagnosis* / Q000821:virology; D002555:Cerebrospinal Fluid / Q000821:virology*; D004279:DNA, Viral / Q000032:analysis / Q000134:cerebrospinal fluid; D006564:Herpesviridae / Q000235:genetics / Q000302:isolation & purification*; D006566:Herpesviridae Infections / Q000134:cerebrospinal fluid / Q000175:diagnosis* / Q000821:virology; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D004279:DNA, Viral