Diagnosis of infection with human herpes viruses in routine laboratory practice.
BACKGROUND
The potential for faster detection of human herpes viruses using PCR compared to other methods is undisputed. However, because of fear of contamination, the clinical implication of nucleic amplification methods in routine laboratories is not widespread. Herpes viruses cause a wide spectrum of diseases and can cause morbidity and mortality in immune-compromised patients. Using real-time PCR, most of the problems associated with PCR (contamination, cumbersome detection, and rather expensive tests) are solved, and a rapid, economical, and--most importantly--closed system is at hand.


METHODS
We evaluated work procedures in our laboratory that enable the routine diagnosis of viral infections with high accuracy and rapid turn-around time. In parallel, inherent problems usually associated with PCR testing, especially cross-contamination could be suppressed to a minimum. The start of the work flow process begins with an automated nucleic acid extraction procedure that yields high quality DNA. A common--internally and externally controlled--PCR program for all six viruses allows rapid sample turn around.


RESULTS
In all, 7500 analyses for human herpes virus infection were performed in the last 5 years. Results for various different specimens were produced within 24 h. Contamination occurred rarely and could be ameliorated easily. The use of internal controls identified rare PCR-inhibited samples. The detection limits for our assays are markedly below the clinically relevant range.


CONCLUSIONS
Our workflow allowed rapid, cost-efficient, and labor saving routine diagnostic detection of viral infections.
Mostafaie
Nazanin
N
Department of Clinical Chemistry, Donauspital, Vienna, Austria.
Huber
Klaus R
KR
Sebesta
Christian
C
Bauer
Kurt
K
Kristoferitsch
Wolfgang
W
Volc-Platzer
Beatrix
B
Kurz
Herbert
H
Klinghofer
Ilse
I
Mirkovic
Borka
B
Hofmann
Jörg
J
Hejtman
Milos
M
Schrattbauer
Karl
K
Krugluger
Walter
W
D019411:Clinical Laboratory Techniques; D006564:Herpesviridae / Q000235:genetics / Q000302:isolation & purification*; D006566:Herpesviridae Infections / Q000175:diagnosis*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*