PMID- 11060066
OWN - NLM
STAT- MEDLINE
DCOM- 20010125
LR  - 20240407
IS  - 0095-1137 (Print)
IS  - 1098-660X (Electronic)
IS  - 0095-1137 (Linking)
VI  - 38
IP  - 11
DP  - 2000 Nov
TI  - Real-time quantitative PCR for human herpesvirus 6 DNA.
PG  - 4042-8
AB  - The diagnosis of human herpesvirus 6 (HHV-6) infection represents a complex issue 
      because the most widely used diagnostic tools, such as immunoglobulin G antibody 
      titer determination and qualitative DNA PCR with blood cells, are unable to 
      distinguish between latent (clinically silent) and active (often clinically 
      relevant) infection. We have developed a new, highly sensitive, quantitative PCR 
      assay for the accurate measurement of HHV-6 DNA in tissue-derived cell 
      suspensions and body fluids. The test uses a 5' nuclease, fluorogenic assay 
      combined with real-time detection of PCR amplification products with the ABI 
      PRISM 7700 sequence detector system. The sensitivity of this method is equal to 
      the sensitivity of a nested PCR protocol (lower detection limit, 1 viral genome 
      equivalent/test) for both the A and the B HHV-6 subgroups and shows a wider 
      dynamic range of detection (from 1 to 10(6) viral genome equivalents/test) and a 
      higher degree of accuracy, repeatability, and reproducibility compared to those 
      of a standard quantitative-competitive PCR assay developed with the same 
      reference DNA molecule. The novel technique is versatile, showing the same 
      sensitivity and dynamic range with viral DNA extracted from different fluids 
      (i.e., culture medium or plasma) or from tissue-derived cell suspensions. 
      Furthermore, by virtue of its high-throughput format, this method is well suited 
      for large epidemiological surveys.
FAU - Locatelli, G
AU  - Locatelli G
AD  - Unit of Human Virology, DIBIT, San Raffaele Scientific Institute, 20132 Milan, 
      Italy.
FAU - Santoro, F
AU  - Santoro F
FAU - Veglia, F
AU  - Veglia F
FAU - Gobbi, A
AU  - Gobbi A
FAU - Lusso, P
AU  - Lusso P
FAU - Malnati, M S
AU  - Malnati MS
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Clin Microbiol
JT  - Journal of clinical microbiology
JID - 7505564
RN  - 0 (DNA, Viral)
RN  - EC 2.7.7.- (Taq Polymerase)
SB  - IM
CIN - J Clin Microbiol. 2001 Aug;39(8):3020-1. PMID: 11499395
MH  - DNA, Viral/*blood
MH  - Herpesviridae Infections/*diagnosis/virology
MH  - Herpesvirus 6, Human/genetics/*isolation & purification/physiology
MH  - Humans
MH  - Polymerase Chain Reaction/*methods
MH  - Reference Standards
MH  - Reproducibility of Results
MH  - Sensitivity and Specificity
MH  - Taq Polymerase/metabolism
MH  - Viral Load
PMC - PMC87539
EDAT- 2000/11/04 11:00
MHDA- 2001/02/28 10:01
PMCR- 2000/11/01
CRDT- 2000/11/04 11:00
PHST- 2000/11/04 11:00 [pubmed]
PHST- 2001/02/28 10:01 [medline]
PHST- 2000/11/04 11:00 [entrez]
PHST- 2000/11/01 00:00 [pmc-release]
AID - 1113 [pii]
AID - 10.1128/JCM.38.11.4042-4048.2000 [doi]
PST - ppublish
SO  - J Clin Microbiol. 2000 Nov;38(11):4042-8. doi: 10.1128/JCM.38.11.4042-4048.2000.