PMID- 23249654
OWN - NLM
STAT- MEDLINE
DCOM- 20130619
LR  - 20211021
IS  - 1743-422X (Electronic)
IS  - 1743-422X (Linking)
VI  - 9
DP  - 2012 Dec 18
TI  - Development and validation of a Q-PCR based TCID50 method for human herpesvirus 
      6.
PG  - 311
LID - 10.1186/1743-422X-9-311 [doi]
AB  - 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(50), 
      and 43% for an infectious units approach using IFA. The average inter-assay CV 
      for quantitative PCR read-out of TCID(50) was 73%, compared to 66%, 25% and 77% 
      for the ocular inspection read-out for TCID(50), IFA read-out of TCID(50)and 
      infectious unit approaches respectively. CONCLUSIONS: The quantitative PCR based 
      read-out of TCID(50)proved to be more robust and easier to interpret than 
      traditional TCID(50)assessment approaches for HHV-6, and therefore it might be 
      considered as an alternative method.
FAU - Gustafsson, Rasmus K L
AU  - Gustafsson RK
AD  - Karolinska Institutet, Department of Clinical Neuroscience, The Multiple 
      Sclerosis Research Group, Center for Molecular Medicine building L8:00, 
      Karolinska University hospital Solna, SE-171 76, Stockholm, Sweden. 
      Rasmus.Gustafsson@ki.se
FAU - Engdahl, Elin E
AU  - Engdahl EE
FAU - Fogdell-Hahn, Anna
AU  - Fogdell-Hahn A
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Validation Study
DEP - 20121218
PL  - England
TA  - Virol J
JT  - Virology journal
JID - 101231645
RN  - 0 (DNA, Viral)
SB  - IM
MH  - Cell Line
MH  - DNA, Viral
MH  - Herpesvirus 6, Human/*genetics/growth & development
MH  - Humans
MH  - *Real-Time Polymerase Chain Reaction
MH  - Reproducibility of Results
MH  - Viral Load/*methods
MH  - Virus Replication
PMC - PMC3546908
EDAT- 2012/12/20 06:00
MHDA- 2013/06/20 06:00
PMCR- 2012/12/18
CRDT- 2012/12/20 06:00
PHST- 2012/07/17 00:00 [received]
PHST- 2012/12/11 00:00 [accepted]
PHST- 2012/12/20 06:00 [entrez]
PHST- 2012/12/20 06:00 [pubmed]
PHST- 2013/06/20 06:00 [medline]
PHST- 2012/12/18 00:00 [pmc-release]
AID - 1743-422X-9-311 [pii]
AID - 10.1186/1743-422X-9-311 [doi]
PST - epublish
SO  - Virol J. 2012 Dec 18;9:311. doi: 10.1186/1743-422X-9-311.