PMID- 28659899
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20200930
IS  - 1664-302X (Print)
IS  - 1664-302X (Electronic)
IS  - 1664-302X (Linking)
VI  - 8
DP  - 2017
TI  - A Novel Method to Titrate Herpes Simplex Virus-1 (HSV-1) Using Laser-Based 
      Scanning of Near-Infrared Fluorophores Conjugated Antibodies.
PG  - 1085
LID - 10.3389/fmicb.2017.01085 [doi]
LID - 1085
AB  - Among several strategies used for Herpes simplex virus (HSV) detection in 
      biological specimens, standard plaque assay (SPA) remains the most reliable 
      method to evaluate virus infectivity and quantify viral replication. However, it 
      is a manual procedure, thereby affected by operator subjectivity, and it may be 
      particularly laborious for multiple sample analysis. Here we describe an 
      innovative method to perform the titration of HSV type 1 (HSV-1) in different 
      samples, using the "In-Cell Western(TM)" Assay (ICW) from LI-COR, a quantitative 
      immunofluorescence assay that exploits laser-based scanning of near infrared 
      (NIR). In particular, we employed NIR-immunodetection of viral proteins to 
      monitor foci of HSV-1 infection in cell monolayers, and exploited an automated 
      detection of their fluorescence intensity to evaluate virus titre. This 
      innovative method produced similar and superimposable values compared to SPA, but 
      it is faster and can be performed in 96 well plate, thus allowing to easily and 
      quickly analyze and quantify many samples in parallel. These features make our 
      method particularly suitable for the screening and characterization of antiviral 
      compounds, as we demonstrated by testing acyclovir (ACV), the main anti-HSV-1 
      drug. Moreover, we developed a new data analysis system that allowed to overcome 
      potential bias due to unspecific florescence signals, thus improving data 
      reproducibility. Overall, our method may represents a useful tool for both 
      clinical and research purposes.
FAU - Fabiani, Marco
AU  - Fabiani M
AD  - Department of Public Health and Infectious Diseases, Sapienza University of 
      RomeRome, Italy.
FAU - Limongi, Dolores
AU  - Limongi D
AD  - San Raffaele Pisana, Istituto di Ricovero e Cura a Carattere Scientifico, 
      Telematic UniversityRome, Italy.
FAU - Palamara, Anna Teresa
AU  - Palamara AT
AD  - San Raffaele Pisana, Istituto di Ricovero e Cura a Carattere Scientifico, 
      Telematic UniversityRome, Italy.
AD  - Department of Public Health and Infectious Diseases, Sapienza University of Rome, 
      Laboratory Affiliated to Istituto Pasteur Italia - Fondazione Cenci 
      BolognettiRome, Italy.
FAU - De Chiara, Giovanna
AU  - De Chiara G
AD  - Institute of Translational Pharmacology, National Research CouncilRome, Italy.
FAU - Marcocci, Maria Elena
AU  - Marcocci ME
AD  - Department of Public Health and Infectious Diseases, Sapienza University of 
      RomeRome, Italy.
LA  - eng
PT  - Journal Article
DEP - 20170614
PL  - Switzerland
TA  - Front Microbiol
JT  - Frontiers in microbiology
JID - 101548977
PMC - PMC5469900
OTO - NOTNLM
OT  - HSV-1
OT  - antivirals
OT  - herpes simplex virus
OT  - immunostaining
OT  - near-infrared fluorescence
OT  - plaque assay
OT  - virus titration
EDAT- 2017/07/01 06:00
MHDA- 2017/07/01 06:01
PMCR- 2017/06/14
CRDT- 2017/06/30 06:00
PHST- 2017/03/14 00:00 [received]
PHST- 2017/05/29 00:00 [accepted]
PHST- 2017/06/30 06:00 [entrez]
PHST- 2017/07/01 06:00 [pubmed]
PHST- 2017/07/01 06:01 [medline]
PHST- 2017/06/14 00:00 [pmc-release]
AID - 10.3389/fmicb.2017.01085 [doi]
PST - epublish
SO  - Front Microbiol. 2017 Jun 14;8:1085. doi: 10.3389/fmicb.2017.01085. eCollection 
      2017.