Development and validation of a Q-PCR based TCID50 method for human herpesvirus 6.

ABSTRACT:

Background

For titer assessment of human herpesvirus 6 (HHV-6), IFA targeting viral proteins or a TCID(50) method with ocular inspection for CPE can be used. These methods rely on the subjective decision of the assessor, obstructing the ability to obtain unanimous results.

Findings

We have developed and validated an alternative TCID(50) read-out approach where infection in the titration culture plate is assessed by viral DNA load change by quantitative PCR. A ten time increase in viral DNA load was determined as cut point for infection since that yielded a maximum correlation with viral protein expression (93%). The average intra-assay CV was 9% for quantitative PCR read-out of TCID(50) compared to 45% for ocular inspection read-out of TCID(50) , 14% for IFA read-out of TCID

SUBMITTER: Gustafsson RK 

PROVIDER: S-EPMC3546908 | biostudies-literature | 2012 Dec

REPOSITORIES: biostudies-literature