PMID- 37368999
OWN - NLM
STAT- MEDLINE
DCOM- 20230731
LR  - 20231104
IS  - 2379-3694 (Electronic)
IS  - 2379-3694 (Linking)
VI  - 8
IP  - 7
DP  - 2023 Jul 28
TI  - Paired Capture and FISH Detection of Individual Virions Enable Cell-Free 
      Determination of Infectious Titers.
PG  - 2563-2571
LID - 10.1021/acssensors.3c00239 [doi]
AB  - Early detection of viruses can prevent the uncontrolled spread of viral 
      infections. Determination of viral infectivity is also critical for determining 
      the dosage of gene therapies, including vector-based vaccines, CAR T-cell 
      therapies, and CRISPR therapeutics. In both cases, for viral pathogens and viral 
      vector delivery vehicles, fast and accurate measurement of infectious titers is 
      desirable. The most common methods for virus detection are antigen-based (rapid 
      but not sensitive) and polymerase chain reaction (PCR)-based (sensitive but not 
      rapid). Current viral titration methods heavily rely on cultured cells, which 
      introduces variability within labs and between labs. Thus, it is highly desirable 
      to directly determine the infectious titer without using cells. Here, we report 
      the development of a direct, fast, and sensitive assay for virus detection 
      (dubbed rapid capture fluorescence in situ hybridization (FISH) or rapture FISH) 
      and cell-free determination of infectious titers. Importantly, we demonstrate 
      that the virions captured are "infectious," thus serving as a more consistent 
      proxy of infectious titers. This assay is unique because it first captures 
      viruses bearing an intact coat protein using an aptamer and then detects genomes 
      directly in individual virions using fluorescence in situ hybridization (FISH); 
      thus, it is selective for infectious particles (i.e., positive for coat proteins 
      and positive for genomes).
FAU - Liu, Yifang
AU  - Liu Y
AD  - Department of Bioengineering, Northeastern University, Boston, Massachusetts 
      02115, United States.
FAU - Potts, Jacob L
AU  - Potts JL
AD  - Department of Bioengineering, Northeastern University, Boston, Massachusetts 
      02115, United States.
FAU - Bloch, Dylan
AU  - Bloch D
AUID- ORCID: 0000-0002-5382-5765
AD  - Department of Bioengineering, Northeastern University, Boston, Massachusetts 
      02115, United States.
FAU - Nian, Keqing
AU  - Nian K
AD  - Department of Bioengineering, Northeastern University, Boston, Massachusetts 
      02115, United States.
FAU - McCormick, Caroline A
AU  - McCormick CA
AD  - Department of Bioengineering, Northeastern University, Boston, Massachusetts 
      02115, United States.
FAU - Fanari, Oleksandra
AU  - Fanari O
AD  - Department of Bioengineering, Northeastern University, Boston, Massachusetts 
      02115, United States.
FAU - Rouhanifard, Sara H
AU  - Rouhanifard SH
AUID- ORCID: 0000-0002-6991-4877
AD  - Department of Bioengineering, Northeastern University, Boston, Massachusetts 
      02115, United States.
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20230627
PL  - United States
TA  - ACS Sens
JT  - ACS sensors
JID - 101669031
SB  - IM
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/methods
MH  - *Viruses/genetics
MH  - Polymerase Chain Reaction
MH  - *Virus Diseases
MH  - Virion
PMC - PMC10621038
OTO - NOTNLM
OT  - aptamer
OT  - infectious titer
OT  - smFISH
OT  - spike antigen
OT  - virus quantification
COIS- The authors declare no competing financial interest.
EDAT- 2023/06/27 19:13
MHDA- 2023/07/31 06:42
PMCR- 2023/11/02
CRDT- 2023/06/27 14:53
PHST- 2023/07/31 06:42 [medline]
PHST- 2023/06/27 19:13 [pubmed]
PHST- 2023/06/27 14:53 [entrez]
PHST- 2023/11/02 00:00 [pmc-release]
AID - 10.1021/acssensors.3c00239 [doi]
PST - ppublish
SO  - ACS Sens. 2023 Jul 28;8(7):2563-2571. doi: 10.1021/acssensors.3c00239. Epub 2023 
      Jun 27.