BACKGROUND: A multiplex-PCR and microarray-based method was designed for detection of eight herpesviruses from clinical specimens. OBJECTIVES: 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. STUDY DESIGN: 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.
BACKGROUND: A multiplex-PCR and microarray-based method was designed for detection of eight herpesviruses from clinical specimens. OBJECTIVES: 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. STUDY DESIGN: 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.
Authors: Anne A Gershon; Judith Breuer; Jeffrey I Cohen; Randall J Cohrs; Michael D Gershon; Don Gilden; Charles Grose; Sophie Hambleton; Peter G E Kennedy; Michael N Oxman; Jane F Seward; Koichi Yamanishi Journal: Nat Rev Dis Primers Date: 2015-07-02 Impact factor: 52.329
Authors: Anne J Jääskeläinen; Sari M Viitala; Satu Kurkela; Satu Hepojoki; Heidi Sillanpää; Hannimari Kallio-Kokko; Tomas Bergström; Jukka Suni; Ale Närvänen; Olli Vapalahti; Antti Vaheri Journal: J Microbiol Methods Date: 2014-03-01 Impact factor: 2.363