PMID- 36268226
OWN - NLM
STAT- MEDLINE
DCOM- 20221024
LR  - 20221222
IS  - 2235-2988 (Electronic)
IS  - 2235-2988 (Linking)
VI  - 12
DP  - 2022
TI  - Development of a high-sensitivity and short-duration fluorescence in situ 
      hybridization method for viral mRNA detection in HEK 293T cells.
PG  - 960938
LID - 10.3389/fcimb.2022.960938 [doi]
LID - 960938
AB  - Coronavirus disease 2019 (COVID-19) is an extremely contagious illness caused by 
      severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Early disease 
      recognition of COVID-19 is crucial not only for prompt diagnosis and treatment of 
      the patients, but also for effective public health surveillance and response. The 
      reverse transcription-polymerase chain reaction (RT-PCR) is the most common 
      method for the detection of SARS-CoV-2 viral mRNA and is regarded as the gold 
      standard test for COVID-19. However, this test and those for antibodies (IgM and 
      IgG) and antigens have certain limitations (e.g., by yielding false-negative and 
      false-positive results). We have developed an RNA fluorescence in situ 
      hybridization (FISH) method for high-sensitivity detection of SARS-CoV-2 mRNAs in 
      HEK 293T cell cultures as a model. After transfection of HEK 293T cells with 
      plasmids, Spike (S)/envelope (E) proteins and their mRNAs were clearly detected 
      inside the cells. In addition, hybridization time could be reduced to 2 hours for 
      faster detection when probe concentration was increased. Our approach might thus 
      significantly improve the sensitivity and specificity of SARS-CoV-2 detection and 
      be widely applied for the high-sensitivity single-molecular detection of other 
      RNA viruses (e.g., Middle East respiratory syndrome coronavirus (MERS-CoV), 
      Hepatitis A virus, all influenza viruses, and human immunodeficiency virus (HIV)) 
      in various types of samples including tissue, body fluid, blood, and water. RNA 
      FISH can also be utilized for the detection of DNA viruses (e.g., Monkeypox 
      virus, human papillomavirus (HPV), and cytomegalovirus (CMV)) by detection of 
      their mRNAs inside cells or body fluid.
CI  - Copyright (c) 2022 Hu, Wang, Uddin, Greene, Hu and Ma.
FAU - Hu, Dailun
AU  - Hu D
AD  - Clinical College, Hebei Medical University, Shijiazhuang, China.
FAU - Wang, Tao
AU  - Wang T
AD  - Telethon Kids Institute, Perth Children's Hospital, Nedlands, WA, Australia.
AD  - Medical School, University of Western Australia, Nedlands, WA, Australia.
FAU - Uddin, Jasim
AU  - Uddin J
AD  - School of Veterinary Medicine, Murdoch University, Murdoch, WA, Australia.
FAU - Greene, Wayne K
AU  - Greene WK
AD  - Medical, Molecular and Forensic Sciences, Murdoch University, Murdoch, WA, 
      Australia.
FAU - Hu, Dakang
AU  - Hu D
AD  - Department of Laboratory Medicine, Taizhou Municipal Hospital, Taizhou, China.
FAU - Ma, Bin
AU  - Ma B
AD  - Medical, Molecular and Forensic Sciences, Murdoch University, Murdoch, WA, 
      Australia.
LA  - eng
PT  - Journal Article
DEP - 20221004
PL  - Switzerland
TA  - Front Cell Infect Microbiol
JT  - Frontiers in cellular and infection microbiology
JID - 101585359
RN  - 0 (RNA, Messenger)
RN  - 0 (Immunoglobulin M)
RN  - 0 (Immunoglobulin G)
RN  - 059QF0KO0R (Water)
SB  - IM
MH  - Humans
MH  - *SARS-CoV-2/genetics
MH  - *COVID-19
MH  - COVID-19 Testing
MH  - Clinical Laboratory Techniques/methods
MH  - RNA, Messenger/genetics
MH  - In Situ Hybridization, Fluorescence
MH  - HEK293 Cells
MH  - Immunoglobulin M
MH  - Immunoglobulin G
MH  - Water
PMC - PMC9577401
OTO - NOTNLM
OT  - HEK 293T cell
OT  - RNA virus
OT  - SARS-CoV-2
OT  - fluorescence in situ hybridization
OT  - mRNA
COIS- The authors declare that the research was conducted in the absence of any 
      commercial or financial relationships that could be construed as a potential 
      conflict of interest.
EDAT- 2022/10/22 06:00
MHDA- 2022/10/25 06:00
PMCR- 2022/10/04
CRDT- 2022/10/21 03:11
PHST- 2022/06/03 00:00 [received]
PHST- 2022/09/20 00:00 [accepted]
PHST- 2022/10/21 03:11 [entrez]
PHST- 2022/10/22 06:00 [pubmed]
PHST- 2022/10/25 06:00 [medline]
PHST- 2022/10/04 00:00 [pmc-release]
AID - 10.3389/fcimb.2022.960938 [doi]
PST - epublish
SO  - Front Cell Infect Microbiol. 2022 Oct 4;12:960938. doi: 
      10.3389/fcimb.2022.960938. eCollection 2022.