Detection of varicella zoster virus DNA in human tissue by standard and nested polymerase chain reaction

Abstract

Abstracts molecules/ml. Most of them were detected by the long probe but not by the short probe. When PCR for the Sor PreS-gene was included it was possible to detect all 24 HBV-positive sera (not shown) by ELOSA. The reliable lower quantification limit for the long probe is 250 molecules/ml and for the short probe 2500 molecules/ml. Surprisingly, chemiluminescence did not produce better qualitative or quantitative results. The data suggest that the usage of several replicates allows relative quantification ~ in most cases. One possible drawback we see is the hybridization efficiency. Six of our positive samples showed great differences between the number of target molecules suggested by agarose gel electrophoreses or by hybridization (Southern blot or ELOSA). All of them contained more than::,! 06 molecules/ml. For these cases and for the samples where the short probe and the long probe gave discordant result (2 cases) we think that competitive PCR will be the method of choice, but in most cases ELOSA with the long probe gives reliable results and is highly sensitive.sponding to expected target copy numbers (5000, 500 and 50 copies/reaction) was employed. The NASBA showed a high reproducibility; the intra-and inter-assay coefficients of variation were < 10%. The results obtained by the amplification of the dilution panel corresponded to the quantification of NASBA QT using the undiluted (5000 copies) HIV-1 RNA standard. So far, the NASBA HIV-1 RNA QT is the only commercially available amplification assay which permits a quantitative detection of HIV-1 RNA. Our results, although preliminary, show that the NASBA allows an accurate and reproducible quantitation of HIV RNA combined with a simple nucleic acid isolation procedure and non-radioactive detection of the amplificates.

Subclinical Reactivation of Varicella-Zoster Virus in Immunocompromised and Immunocompetent Individuals

“…After primary infection, the virus establishes a life-long latent infection in dorsal root ganglion cells. Several times it has been shown by PCR that VZV DNA is detectable in ganglia [1][2][3]. The infection rate of VZV reaches 60-70% at the end of the first decade of life and about 90% at the end of the second decade [4].…”