Comparison of nested PCR and real time PCR of Herpesvirus infections of central nervous system in HIV patients.
BACKGROUND
Molecular detection of herpesviruses DNA is considered as the reference standard assay for diagnosis of central nervous system infections. In this study nested PCR and real time PCR techniques for detection of Herpes simplex virus type 1 (HSV-1), Cytomegalovirus (CMV) and Epstein-Barr virus (EBV) in cerebrospinal fluid of HIV patients were compared.


METHODS
Forty-six, 85 and 145 samples previously resulted positive for HSV-1, CMV and EBV by nested PCR and 150 randomly chosen negative samples among 1181 collected in the period 1996-2003 were retrospectively reassessed in duplicate by real time PCR and nested PCR.


RESULTS
Samples giving positive results for CMV, HSV-1 and EBV with nested PCR were positive also with real time PCR. One of the negative samples resulted positive for HSV and one for EBV. Real time PCR showed comparable sensitivity and specificity vs nested PCR.


CONCLUSIONS
Real time PCR proved to be a suitable method for diagnosis of herpesvirus infections in CNS, showing comparable sensitivity and being less time consuming than nested PCR.
Drago
Lorenzo
L
Laboratory of Clinical Microbiology, L. Sacco Teaching Hospital and Department of Clinical Sciences L, Sacco, University of Milan, Via GB Grassi74, 20157 Milan, Italy. lorenzo.drago@unimi.it
Lombardi
Alessandra
A
De Vecchi
Elena
E
Giuliani
Giuseppe
G
Bartolone
Rosaria
R
Gismondo
Maria Rita
MR
D017088:AIDS-Related Opportunistic Infections / Q000175:diagnosis* / Q000821:virology; D020805:Central Nervous System Viral Diseases / Q000175:diagnosis* / Q000821:virology; D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification; D003586:Cytomegalovirus Infections / Q000175:diagnosis / Q000821:virology; D004279:DNA, Viral / Q000134:cerebrospinal fluid; D020803:Encephalitis, Herpes Simplex / Q000175:diagnosis / Q000821:virology; D020031:Epstein-Barr Virus Infections / Q000175:diagnosis / Q000821:virology; D006564:Herpesviridae / Q000235:genetics / Q000302:isolation & purification*; D006566:Herpesviridae Infections / Q000175:diagnosis* / Q000821:virology; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000302:isolation & purification; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D008587:Meningitis, Viral / Q000175:diagnosis / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012189:Retrospective Studies; D012680:Sensitivity and Specificity
D004279:DNA, Viral