Improved multiplex-PCR and microarray for herpesvirus detection from CSF

BackgroundA multiplex-PCR and microarray-based method was designed for detection of eight herpesviruses from clinical specimens.ObjectivesTo 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.Study designA 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.ResultsNew method was more sensitive for detection of HSVs and both two new detection oligonucleotides for VZV functioned well at low levels of viral DNA.ConclusionsThe revised HSV-PCR and new HSV- and VZV-oligonucleotides were found to function well and be more sensitive, thereby increasing reliability of the method.