Diagnosis of infection with human herpes viruses in routine laboratory practice

Abstract

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

Occurrence of viral DNA in paired samples of corneal rim and cornea preservation fluid

“…In this study, detection of adenovirus DNA was done with qualitative method by means of probe and starters complementary to the gene encoding the hexon protein—representing a conservative region of human adenoviruses. A hydrolysis probe of the TaqMan type, labeled with fluorochrome JOE/560, was used [Mostafaie et al, ; Rynans et al, ]. Limit of HAdV detection was 256 copies/ml.…”