PMID- 38421570
OWN - NLM
STAT- Publisher
LR  - 20240229
IS  - 1559-0291 (Electronic)
IS  - 0273-2289 (Linking)
DP  - 2024 Feb 29
TI  - A Method Based on a Modified Fluorescence In Situ Hybridization (FISH) Approach 
      for the Sensing of Staphylococcus aureus from Nasal Samples.
LID - 10.1007/s12010-024-04892-9 [doi]
AB  - Staphylococcus aureus is a major source of bacteremia and develops several 
      complications, causing high morbidity and mortality. Rapid identification and 
      detection of these bacteria have become an important issue for biomedical 
      applications. Herein, an optical method based on a modified fluorescence in situ 
      hybridization (FISH) approach has been established using DNA hybridization 
      technology for the swift detection of pathogenic S. aureus from clinical samples. 
      The platform was constructed with single-stranded genomic DNA and microbial 
      colony by directly immobilizing in agarose-polyvinyl alcohol (AG-PVA) hydrogel on 
      the surface of a glass slide. The probe was based on an elongation factor 
      encoding the tuf gene, which binds with equal affinity to single-stranded DNA 
      targets as well as surface proteins on microbial cells. The probe was labeled 
      with MFP488 fluorophore having excitation wavelength 501 nm. The hybridization of 
      the labeled probe with the target DNA and surface proteins was carried out under 
      optimal FISH conditions, and the detection of bacteria was based on temporary 
      field excitation of the labeled probe under a fluorescence microscope. Positive 
      hybridization signals were detected by high fluorescence intensity. In comparison 
      to genomic DNA, robust signals were observed with microbial cells, perhaps due to 
      the moonlighting effect of the elongation factor Tu (Ef-Tu) expressed on the 
      surface of bacterial cells. The applicability of the developed platform was 
      tested on pediatric nasal samples, and results were verified with real-time qPCR. 
      The designed platform is stable and sensitive, and after detailed optimization, a 
      portable structure for on-site detection of pathogenic bacteria from clinical 
      samples can be produced.
CI  - (c) 2024. The Author(s), under exclusive licence to Springer Science+Business 
      Media, LLC, part of Springer Nature.
FAU - Sami, Amtul Jamil
AU  - Sami AJ
AD  - School of Biochemistry and Biotechnology, University of the Punjab, Lahore, 
      54590, Pakistan. amtuljamilsami@yahoo.com.
AD  - Center for Biosensor Research and Development, University of the Punjab, Lahore, 
      54590, Pakistan. amtuljamilsami@yahoo.com.
FAU - Bilal, Sehrish
AU  - Bilal S
AD  - School of Biochemistry and Biotechnology, University of the Punjab, Lahore, 
      54590, Pakistan.
AD  - Department of Biochemistry, Gulab Devi Educational Complex, Lahore, 54600, 
      Pakistan.
FAU - Alam, Sadaf
AU  - Alam S
AD  - School of Biochemistry and Biotechnology, University of the Punjab, Lahore, 
      54590, Pakistan.
FAU - Khalid, Madeeha
AU  - Khalid M
AD  - School of Biochemistry and Biotechnology, University of the Punjab, Lahore, 
      54590, Pakistan.
AD  - Department of Biochemistry, University of Okara, Okara, 56300, Pakistan.
FAU - Mangat, Hammad Ahmad
AU  - Mangat HA
AD  - Faryal Dental College Lahore, Sharaqpur, Lahore, 39460, Pakistan.
LA  - eng
PT  - Journal Article
DEP - 20240229
PL  - United States
TA  - Appl Biochem Biotechnol
JT  - Applied biochemistry and biotechnology
JID - 8208561
SB  - IM
OTO - NOTNLM
OT  - Staphylococcus aureus
OT  - Agarose
OT  - Biosensor
OT  - Fluorescence
OT  - Hydrogel
OT  - PVA
EDAT- 2024/02/29 12:46
MHDA- 2024/02/29 12:46
CRDT- 2024/02/29 11:14
PHST- 2024/02/12 00:00 [accepted]
PHST- 2024/02/29 12:46 [medline]
PHST- 2024/02/29 12:46 [pubmed]
PHST- 2024/02/29 11:14 [entrez]
AID - 10.1007/s12010-024-04892-9 [pii]
AID - 10.1007/s12010-024-04892-9 [doi]
PST - aheadofprint
SO  - Appl Biochem Biotechnol. 2024 Feb 29. doi: 10.1007/s12010-024-04892-9.