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Towards Point of Care Diagnostics: Visual Detection of Meningitis Pathogens Directly from Cerebrospinal Fluid
Author(s) -
Kovtunov Evgeny A.,
Shkodenko Liubov A.,
Goncharova Ekaterina A.,
Nedorezova Daria D.,
Sidorenko Sergey V.,
Koshel Elena I.,
Kolpashchikov Dmitry M.
Publication year - 2020
Publication title -
chemistryselect
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.437
H-Index - 34
ISSN - 2365-6549
DOI - 10.1002/slct.202003869
Subject(s) - amplicon , polymerase chain reaction , neisseria meningitidis , cerebrospinal fluid , microbiology and biotechnology , molecular diagnostics , biology , bacteria , streptococcus pneumoniae , meningitis , virology , antibiotics , medicine , bioinformatics , gene , genetics , neuroscience , psychiatry
Rapid and cost‐efficient methods for diagnostics of infectious diseases can expedite prescription of adequate treatment, thus shortening the patients’ recovery and reducing adverse side effects. In this study, we developed a procedure for visual detection of bacterial pathogens Gram‐negative Neisseria meningitidis and Gram‐positive Streptococcus pneumoniae directly from human samples of cerebrospinal fluid (CSF). The assay uses hybridization probes that can efficiently recognize folded single‐stranded DNA analytes due to their split design. The assay includes the following stages: PCR amplification of N. meningitidis and S. pneumoniae targeted sequences; λ exonuclease treatment of the polymerase chain reaction (PCR) amplicons, and detection of the amplicons by hybridization probes with visual signal output. The assay requires 2.0 h with hands‐on time of about 40 min, and a regular PCR thermal cycler. It detects down to 10 bacteria in 2 μL of cerebrospinal fluid. The assay works for both gram‐positive and gram‐negative bacteria and does not require optimization of PCR conditions. This development creates a basis for the point‐of‐care diagnostics of bacterial meningitis with visual signal output.