Simultaneous detection of 6 human herpesviruses in cerebrospinal fluid and aqueous fluid by a single PCR using stair primers.
A Herpes Consensus allows the simultaneous detection of 6 human herpesviruses: herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2), human cytomegalovirus (HCMV), varicella-zoster virus (VZV), Epstein-Barr virus (EBV), and human herpes virus 6 (HHV-6). This technique was used first to examine retrospectively 100 DNA extracts from 95 CSF and 5 aqueous fluids, prepared by treatment by saturated NaCl followed by ethanol precipitation (n = 63) or by simple boiling (n = 37) and stored at -80 degrees C, and secondly to test prospectively 38 CSF samples for which two DNA extracts were prepared with commercially available DNA extraction kits. In all cases, the results were compared with those of an "in-house" PCR. Concordant results between both PCR and the Herpes Consensus techniques were obtained in 61 of 63 DNA extracts prepared by treatment by saturated NaCl (97%) and in only 31 of 37 boiled samples (84%). Both commercially available methods of DNA extraction examined appear to be suitable for Herpes Consensus PCR, although they cannot remove completely PCR inhibitors that must be sought in case of negative results. This preliminary study shows that the Herpes Consensus method should be of value for rapid diagnosis of herpesvirus infections on condition that it is performed on purified DNA extracts.
Bouquillon
C
C
Laboratoire de Virologie, CHU de Lille, France.
Dewilde
A
A
Andreoletti
L
L
Lambert
V
V
Chieux
V
V
Gerard
Y
Y
Lion
G
G
Bocket
L
L
Wattre
P
P
D001082:Aqueous Humor / Q000821:virology*; D002494:Central Nervous System Infections / Q000821:virology; D002555:Cerebrospinal Fluid / Q000821:virology*; D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification; D004279:DNA, Viral / Q000032:analysis*; D000431:Ethanol; D015828:Eye Infections, Viral / Q000821:virology; D005615:Freezing; D006564:Herpesviridae / Q000235:genetics / Q000302:isolation & purification*; D006566:Herpesviridae Infections / Q000821:virology; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000302:isolation & purification; D018258:Herpesvirus 2, Human / Q000235:genetics / Q000302:isolation & purification; D014645:Herpesvirus 3, Human / Q000235:genetics / Q000302:isolation & purification; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification; D015654:Herpesvirus 6, Human / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D012965:Sodium Chloride
D004279:DNA, Viral; D000431:Ethanol; D012965:Sodium Chloride